DETECTION AND SEPARATION

FIELD OF THE INVENTION

[0001] The invention relates to the analysis and processing of microorganisms, bio-molecules, and other small particles. In particular, the invention relates to systems, methods, components, and controllers for the separation, identification, classification, and containment of bacteria and other microbes, and other particles, including nanoparticles.

BACKGROUND OF THE INVENTION

[0002] Microbes are the largest contributors to earth's biodiversity. It is estimated that 85% - 99% of environmental bacteria have not yet been cultivated, much less put to productive use. Moreover, challenges to identification and analysis of the bulk of microbes are significant. For example, currently-available enumeration technologies such as massive DNA sequencing, fluorescent in-situ hybridization, phylochip etc. are all nucleotide-based. This and other limitations mean that sequencing errors are unacceptably high, and biologically genome mosaicism can occur, due to the possibility of horizontal gene transfer. Moreover, each of these techniques is destructive, so that follow-up through in-depth characterization, cultivation, and experimentation become impossible.

[0003] Such issues in the identification and understanding of microbes are of significant importance to future humankind, and they suggest the desirability of improving processes for detecting, separating, and otherwise processing microbes. However, the same is true of a very broad range of other small particles, including biomolecules and a wide variety of nanoparticles.

[0004] In view of the above and other shortcomings, many of which will be apparent to those skilled in the relevant arts upon a review of the following disclosure, it would be advantageous to provide, among other things, improved separators, detectors, and sorting and collecting systems, and systems comprised wholly or partly or parts or combinations thereof, and processes and stored machine- readable instruction sets adapted for use by such systems and components in identifying, separating, and detecting bacteria and other microbes, biomolecules, and other small particles as described below. Among other improvements, it would be desirable to provide such systems and components configured for non-destructively separating, detecting, identifying, and otherwise processing such particles.

SUMMARY OF THE INVENTION

[0005] We have developed new systems, methods, controllers, and other devices and components, including machine-readable instruction sets, useful in the separation, identification, classification, containment (or 'partitioning'), and other processing of bacteria and other microbes and bio-molecules, and other particles, including nanoparticles. Such innovations are useful, for example, in advancing the new sciences of bactomography (advancement of the study of bacteria) and micromography (advancement of the study of microorganisms, including for example fungi). Among other advantages, the invention enables mixtures or combinations of unknown microbes to be reliably, efficiently, and non-destructively separated, or partitioned, and sorted for identification and other analysis.

[0006] For purposes of this specification, a 'particle' is any object having major dimensions (e.g., greatest diameter or width) measuring from sub-microns to tens of microns. Such particles include, but are not limited to, molecules (including all bio- molecules), bio-particles, amino acids, and organic or otherwise inert particles. For processing in accordance with the various aspects of the invention, such particles can be, but are not necessarily, suspended in gasses or liquids, or provided in or with solid mixtures.

[0007] The disclosure herein refers, in various aspects and embodiments, to 'hybrid' systems and/or components, including for example hybrid detection and separation systems, and components thereof. In such usage, the term 'hybrid' refers to the employment of multiple methods, and types of methods, to enhance the efficiency, accuracy, consistency, and general performance of the systems. For example, as will be seen below, hybrid systems, subsystems, and components as contemplated herein can apply multiple physical principles, and thereby take advantage of various physical attributes of particles, to separate, detect, identify, contain (partition), and/or otherwise process them. Hybrid systems and components can also refer to systems and components that are tuned, optimized, or otherwise suitable for use in processing a wide range of incoming sample species. Such hybrid systems can, for example, include separation systems that combine the use of centrifugal and electrical forces to separate molecules and particles according to their size and charge.

[0008] In one aspect, the invention provides systems for separating and partitioning particles, including bacteria and other microbes. Such systems can comprise one or more separation stages, one or more detection stages, and one or more collection stages. In further aspects, the invention provides improved hybrid and/or single-purpose separation, detection, and collection stages suitable for use in such systems, and in systems of other types.

[0009] In various aspects and embodiments, systems and components in accordance with the invention include multiple stages. For example, the inventors have found that separation components which combine the use of multiple, hybrid separators, e.g., inertial (e.g., centrifugal) and electric-field separators, yield a number of advantages, which can be tailored by reasoned selection of the types of separators used. Similarly, the use of hybrid and other multi-stage detection and containment systems, including systems that provide multiple, parallel processing paths, have been found to be highly advantageous. Such hybrid and/or other multistage separation components can, for example, increase the accuracy, reliability, and consistency of identification, separation, and partitioning processes, reduce processing time, and enable the use of compact system configurations. Design flexibilities introduced through the use of multi-stage systems, optionally including hybrid subsystems or components, enable the invention to be implemented in the form of large-scale, fast, and very high throughput systems, suitable for example for use by analytic laboratories and large-scale, centralized processing, as well as in the form of physically smaller systems suitable for use in clinics, medical offices, and other specialized analysis centers. The use of hybrid or multi-stage separation components can also, for example, enable the separation of particles according to a wide variety of combinations of size, mass, shape, and electrical charge.

[0010] Thus, it will be appreciated by those skilled in the relevant arts that both scalability and the ability to process analyses at variable speeds are among the numerous advantages offered by the invention; and open up new possibilities in environmental, medical, pharmaceutical, agricultural, food processing, research, industrial, and a wide variety other applications. [0011] A further significant advantage offered by the invention is the ability to provide extremely precise analyses, and, through the use of multiple processing channels and other improvements disclosed herein, to do so at greatly improved processing speeds.

[0012] Thus it will be appreciated that in various aspects and embodiments, systems and detection components in accordance with the invention can comprise multiple detection stages and/or multiple throughput channels. The use of multiple throughput channels, or conduits, for example, can provide significant increases in the speed of processing, particularly where larger numbers of microbes or other particles are to be processed. It can also help in the optimization of analysis logistics.

[0013] The use of hybrid detection systems, comprising detection components of multiple types, can provide refinements in the detection and processing of particles according to a wide variety of characteristics. Examples of such hybrid detection components, or stages, include vacuum-based devices and optical manipulation devices. As those skilled in the relevant arts will appreciate, such different types of detection components, or devices, can be used either alone or in any desired combination(s).

[0014] In further aspects, the invention provides a number of mechanical, electrical, and other improvements in individual stages, including separators, detectors, and other system components, including for example the configuration(s) of such devices, and the sequence(s) in which the devices are applied in various forms of particle processing. As previously noted, these improvements enable a wide variety of implementations of systems in accordance with the invention, from relatively large systems intended for industrial usage to smaller systems intended for clinics and specialist laboratories.

[0015] For example, separation components in accordance with the invention can comprise a plurality of centrifugal separation chamber designs, including but not limited to configurations comprising vertical, horizontal and/or center axis chamber rotation, and a variety of sidewall profiles adapted for specific purposes. In addition, or alternatively, operation sequences of rotors can be tailored for dynamic optimization between retention time and separation power under various analysis conditions.

[0016] Further component improvements include specialized configurations of charged plates for separation, detection, and/or collection devices, and configuration of capillaries and other flow channels, or conduits, etc. in separation and other stages. Further improvements include the use of gravity, thermodynamics, and electromagnetic fields, separately or in combination, in separation stages; as well as multi-channel parallel or otherwise arrayed channels, conduits, and/or cells in detection components for high throughput detection & to accommodate / compensate for multi-stage separation. Electrical potentials applied to electric field generators used in the invention can include two-dimensional (2D) fields, gradient and/or multi-zone fields suitable for use in conjunction with parallel and other multichannel capillary designs, of cylindrical and other tubular design. Operation sequence variations, such as alternating field application in separation, detection, and/or collection stages, can be used to advantage in a variety of specific circumstances.

[0017] Among the many significant advantages enabled by the invention and disclosed herein is the non-destructive detection, separation, and other processing of microbes and other particles, and particularly those of widely varying size and other characteristics. Such advantages are accomplished, for example, through the use of high-resolution optical devices.

[0018] In various further aspects and embodiments, the invention provides improvements in the manipulation and control of microbes and other particles. Such improvements include the use of any or all of optical, mechanical, or fluidic (i.e., liquid and/or gaseous) means to facilitate and increase the quality, effectiveness, and efficiency of detection and/or identification of microbes and other particles. For example, detection components in accordance with the invention can incorporate pluralities of excitation devices, sensor arrays, and computational devices, including highly specialized configurations thereof, operated in various combinations, to capture and interpret light of transmitted, reflected, diffracted, or modulated through particle interactions during processes of the type disclosed herein. Identification of particles, for example, can be accomplished through interpretation of light properties such as intensity, frequency, phase, polarization, and/or one- or multi-dimensional reflection, refraction, diffraction, and modulation patterns caused by the impingement on or other interaction of light with particles as they are manipulated in accordance with our disclosure. Other forms of detection method suitable for use in accordance with the invention include crystallography, Raman spectroscopy, and diffraction spectroscopy.

[0019] As a specific example, excitation through the use of pluralities of lasers controlled through the use of micro-controllers and the variation of laser power, spectrum, frequency, phase, polarization, and other characteristics can be used to very rapidly, finely, and accurately control movement and/or identification of particles.

[0020] A further aspect of the invention is the implementation of a variety of return (or "feedback") loops, or paths, in many system configurations. Such return systems can reduce or eliminate a variety of potential inefficiencies, including, for example, the possibilities of losses of time and/or sample due to the need to transfer incompletely-analyzed samples to separate containment vessels in order to return them to the start, and/or other desired point(s), in an analysis process.

[0021] In further aspects and embodiments, the invention provides data processors; persistently-stored, machine-readable and/or executable data structures representing data and/or coded instructions useable by such processors; and methods of using each of the foregoing, to control and otherwise implement separation, detection, containment, and other processes disclosed herein. Such processes include, for example, the implementation of processes controlled at least partly through the use of non-deterministic data-sets in order to identify, classify, and/or sort particle samples based on a wide variety of physical characteristics, including mass, size, and electrical charge.

[0022] In further aspects and embodiments, the invention provides processes of using systems, components, and other devices (including data processors) and data structures as described herein. In addition to the separation, detection, identification, and containment of microbes and other particles, such processes include, for example, cleaning, tuning, and conditioning processes to be used in setting up, maintaining, and using such devices. [0023] For example, a cleaning and conditioning process in accordance with the invention can include processes for self-contained cleaning and conditioning processes at individual stage(s) (or "section(s)"), or at the level of a complete system, including separation, detection, and containment sections (i.e., "complete chain" cleaning processes. Any or all such processes may be activated automatically, as part of power-up or power-down sequences, for example. Such conditioning processes may be configured, for example, to maximize or otherwise optimize or maintain process efficiencies through the treatment of various potential particle contact surfaces through the use of controlled rotation speeds, flow path surface coatings, flow speeds and/or paths, and the application of desired electrical potentials, etc. Alternatively, or in addition, various components of the systems can be removable, so that, for example, they may simply be discarded and replaced, cleaned and re-used, etc.

[0024] By all of the above means the invention provides significant improvements over the prior art. As noted above, for example, the many advantages offered by the invention, which are numerous, include but are not limited to:

• High throughput: using multiple particle processing channels and/or hybrid separation and detection components, results can be available in tens of minutes, or less, rather than days;

• No culture needed: Samples can be identified without the need of growing sample sizes through culturing methods, which introduce further processing delays, particularly at the front end. This improvement can save days or weeks in identification and other analysis processes;

• High resolution capability: Bio-molecules and other particles can be detected down to sub-micron size;

• Optimized identification processes: (1) Built-in cycling systems minimize the need for prior knowledge of sample content; (2) Samples may be contained within equipment during multiple cycles to minimize contamination in laboratories, etc.;

• Versatile design architecture: Amenable to portable equipment as well as integration into in-line monitoring facilities; • Non-destructive partitioning, identification, containment, and other processing of bacteria and other biological or non-biological particles.

[0025] The extraordinarily wide variety of applications and improvements enabled by the wide variety of configurations and processes enabled by the disclosure herein will be apparent to those skilled in the relevant arts, once they have been made familiar with this disclosure. Such applications and improvements are not limited by the many specific examples and embodiments described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Various aspects and embodiments of the invention are illustrated in the accompanying drawings, which are meant to be exemplary and not limiting, and in which like references are intended to refer to like or corresponding parts.

[0027] Figure 1 is a schematic diagram of an embodiment of a system for the separation, detection, sorting, containment, and other processing of bacteria, microbes, and other particles in accordance with the disclosure.

[0028] Figure 2 is a schematic diagram of an embodiment of a multi-stage hybrid particle separation system in accordance with the disclosure.

[0029] Figures 3 - 9 are schematic diagrams of embodiments of inertial separation systems, and components thereof, in accordance with the disclosure.

[0030] Figures 10 - 18 are schematic diagrams of embodiments of electric field separation systems, and components thereof, in accordance with the disclosure.

[0031] Figure 19 is a schematic diagram showing illustrative hypothetical comparisons of results of the use of single-purpose and hybrid separation systems in accordance with the disclosure.

[0032] Figures 20 - 25 are schematic diagrams of embodiments of particle detection systems and components thereof, in accordance with the disclosure.

[0033] Figures 26A-26D are schematic diagrams of particle sorting and collection systems, and components thereof, in accordance with the disclosure.

[0034] Figures 27A-27B are schematic diagrams of sample flow diversion devices in accordance with the disclosure. DESCRIPTION OF EMBODIMENTS

[0035] Preferred embodiments of methods, systems, and apparatus according to the invention are described through reference to the drawings.

[0036] In various aspects and embodiments the invention provides systems, methods, components, and controllers useful in the separation, identification, classification, containment, and other processing of particles, including bacteria and other microbes. Figure 1 is a schematic diagram of such a system 1000. In the embodiment shown, system 1000 comprises one or more of each of sample inlets 1002, separation (sub-)systems or stages 100, detection (sub-)systems or stages 200, collection and storage (sub)systems or stages 300, sample outlets 1009, and controller (sub)systems 400.

[0037] Sample inlets 1002 are configured to accept samples in any desired or otherwise appropriate form(s), which may include solid, gaseous, and/or liquid, so far as the sample can be suspended in a liquid system.

[0038] Separation (sub-)systems or stages 100 make use of any desired physical property(ies), as described herein, to separate samples comprising particles of multiple types (e.g., biological samples comprising multiple types of proteins and/or other substances) for detection and identification by detection (sub-)systems or stages 200, as explained in greater detail below.

[0039] Detection (sub-)systems or stages 200 make use of any desired physical property(ies), as described herein, to evaluate properties of particles separated by separation stage(s) 100, generally in order to allow the particles to be uniquely identified, and their properties evaluated, as explained in greater detail below.

[0040] Collection, storage, or partitioning (sub)systems or stages 300 make use of any desired physical property(ies), as described herein, to provide means for containing separated and evaluated particles, for any further desired use(s). Stage(s) 300 can, for example, direct or otherwise move particles to storage containers via one or more sample outlets 1009, and/or provide feedback loops to any desired processing point of a system 1000, for further refinement or analysis. For example, feedback loops can be configured to selectively return all or any part of an evaluated sample to any desired point(s) of stage(s) 100, 200. [0041] Controller (sub)systems 400 provide any desired or otherwise useful data and signal processing systems 402, volatile or persistent memory(ies), storage devices, or other media 404, input devices 406, and/or output devices 408, to enable full or partially automatic control of any components of a system 1000, and/or any processes performed thereby. This can include, for example, such micro-controllers and/or other components adapted for the control of electrical, optical, and physical valves and switches, liquid and/or gas temperatures and pressures, flow, etc., as would be required or otherwise desirable in implementing the processes described herein. For example, suitably-configured microcontrollers can be incorporated in specially-configured firmware for component and system-level coordination, control etc.

[0042] As will be appreciated by those skilled in the relevant arts, each of (sub)systems or stages 100, 200, 300, 400, and their various components, may comprise any device(s) compatible with the purposes disclosed herein. Wide varieties of such devices are now available, either off the shelf or by modification or fabrication in accordance with the purposes and principles disclosed herein; doubtless further suitable devices will developed hereafter.

[0043] In particular, each of stage(s) 100, 200, 300 can employ any of a wide variety of techniques, making use of any suitable physical (including electrical and/or magnetic) property(ies) of a sample to partition or otherwise separate, detect, and contain particles of a sample population. For example, such systems can incorporate any or all of centrifugal or other inertial devices, electrical charge manipulation devices, and in some cases light-based (optical) devices. Many preferred systems in accordance with the invention use multiple channel and non-destructive inertial and charge-based separation techniques; optical detection to quickly obtain particles' type, size, chemical signature, population count and exact species; and inertial, electrical/magnetic, and/or optical principles to sort or partition the particles for analysis, containment, or transport, and to initiate feedback cycle processes for finer- resolution detection.

[0044] Each of stage(s) 100, 200, 300 may be connected to each other via one or more physical communications means or connections 2002, 3002, etc. Similarly, sample inlet(s), outlet(s), and return or feedback loops 1002, 1009 may be of any design(s) and configuration(s) suitable for the intended application(s) of a system 1000, and may employ any desired physical properties of samples to be introduced and released to containment.

|0045] Figure 2 is a schematic diagram of an embodiment of a separation

(sub)system, or stage, 100 in accordance with the invention. In the embodiment shown, a separation stage 100 comprises one or more of each of sample inlets 1002, 1103, inertial stages 1100, electric-field flow separators 1200, outlets 2002, and interstage flow or connection channels or conduits 1 103, 1203, and any other components suitable for implementing the purposes described herein. A gaseous, liquid, plasma, or solid sample 702 (see e.g. Figure 3) is introduced at inlet 1002 and optionally passed to component inlet(s) 1103, for passage to inertial stage(s) 1100. After initial separation based wholly or partially on centrifugal and/or other inertial properties of the sample and its constituent particles by inertial stage(s) 1100, the sample, or any desired portion(s) of it, are passed to one or more electric-field flow separators (EFSs) 1200 via one or more inter-component connecting channels 1203. EFS separator(s) 1200 can employ charge plates, for example array(s) of capillaries disposed between electrodes, Halbach arrays, and/or other electro-magnetic field generators suitable for a desired application, etc. Separated samples may be passed to detection and/or collection stage(s) 200, 300 via inter-stage flow channel(s) 2002.

|0046] Figure 3 is a schematic diagram of an embodiment of an inertial separation stage 1100 in accordance with various aspects and embodiments of the invention, in the form of a centrifugal elutriation device 1100, 1 150. In the embodiment shown, device 1100, 1150 comprises a body 1107 comprising a chamber wall 1 101 , base 1102, and cap 1105 forming a chamber 1 120, which contains sample 702 shown in the form of a solution or suspension. The body 1107 is adapted to rotate about an axis 1 110 defined by inlet channel (e.g., a tube or other conduit) 1103. Inlet 1103 can further provide lateral support for the body via inlet interface 1 125, which may for example include ball or other bearings. The body 1107 may be rotated by a motor, transmission, or other means engaging any suitable portion of the body 1107, including for example any one or more of base 1102, cap 1105, and wall 1101. Alternatively or in addition, body 1107 may rotated by a motor or transmission engaging inlet 1103. By rotating the body 1 107 about the axis 11 10, for example as shown by arrow 1 111 , at a sufficiently high speed, separator 1100 can cause desired portion(s) to flow upward along interior surface(s) of chamber wall(s) 1101 toward outlet(s) 1 104 which communicate with any or all of further separation stage(s) 1100, 1200, detection stage(s) 200, and collection stage(s) 300. Outlet(s) 1104 may be formed by continuous or segregated annular opening(s) in cap 1105, chamber wall 1 101 , etc.

[0047] As will be appreciated by those skilled in the relevant arts, centrifugal separator(s) 1150 are only one type of inertial separator that is suitable for use in implementing the invention. While centrifugal separators are among the most preferred forms of inertial separators, based on their ability to provide rapid, continuous, and very fine control of physical separation of particles, inertial separators employing curved flow paths, such as helical capillaries, may also be advantageously incorporated in various embodiments of the invention, as discussed in greater detail below. Moreover, the inducement of laminar and/or other flow patterns, profiles, and regimes in particles suspended in fluids while passing through capillaries, as described further below can also be used to improve separation of particles. Thus, it may be seen that in various applications, other forms of inertial separation systems may serve, as alternatives or in addition to centrifugal separators. Other types of systems operating on inertial principles may also be used. For example, in some applications in which relatively low throughput / high latency in processing acceptable, time-of-flight particle separators will serve.

[0048] As will be appreciated by those skilled in the relevant arts, selective control of the rotating speed of a centrifugal separator 1 100, 1150 via controller 400 can cause particles of different masses or densities to flow along the chamber wall(s) 1101 at different rates, so that they may be selectively removed via outlet(s) 1104. Additional degrees of control may be obtained by orienting the separator 1100 such that axis 11 10 is vertical, horizontal, or variable.

[0049] In operation, solution or other sample 702 can be injected into chamber

1120 via either or both of outlet(s) 1002, 1203, as shown by arrows 1103e, 1103f, optionally while the separator 1100 is rotating. As the separator rotates, smaller cells or particles, which may for example be characterized by relatively low sedimentation levels, will be subjected to a net force (which may for example comprise some or all of centrifugal, gravitational, and drag forces) that is higher than the net force(s) experienced by, for example, larger cells or particles with, for example, higher sedimentation levels. As a result, relatively smaller cells or particles will tend to move more quickly toward outlet(s) 1104 than larger cells or particles. Cells (particles) which experience balanced forces will tend to remain at their locations. Increasing the rotational speed and/or the inward flow of sample 702 through inlet 1103 will elute the smaller particles first; larger cells (particles) can be eluted by further increasing the rotational speed and/or the flow of sample 702, either by increasing input flow via inlet 1 103, by use of selected chamber surface coatings on walls 1101 , inlet(s) 1103, etc., and/or the rotational speed of the chamber 1107.

[0050] Figures 4 - 5 are schematic diagrams showing embodiments of walls

1101 of centrifugal separators 100, 1150 in accordance with the invention. In Figure 4a, it is shown that wall(s) 1101 may be straight, or that their relative slope(s) Θ relative to axis 1 110 of rotation may be varied, in order to control absolute or relative elutriation speeds, depending upon the rotational speed of the separator, the physical properties of the sample 702, including mass, size, viscosity, density, etc. Constant-slope embodiments 1101a such as that shown in Figure 4 can be relatively easier, and therefore less expensive, to fabricate, than other configurations. They can also be used in such fashion as to provide relatively high separation efficiencies, through for example careful control of rotational speeds of the separator 1100, 1150.

[0051] In Figure 4B it is shown that wall(s) 1101 may be curved, either in convex shapes (as shown), concave (not shown), or both (i.e., complex) (not shown), such that slopes Θ of such walls 1101 b relative to axis 1 110 of rotation may be tailored to achieved any desired results. In general, it has been found that carefully defined curvature of chamber walls 1 101 b can be used to selectively increase or decrease sample retention time(s), though in some circumstances this can result in relatively lower separation efficiencies.

[0052] In Figure 5, it is shown that wall(s) 1101 may incorporate grooves, channels, wedges, ridges, or other discontinuities 1130 in order to help guide sample particles 702 toward outlet(s) 1203 in various ways for various purposes. Such features 1130 may be of any desired configuration(s) - straight, curved (e.g., helical), etc., and may be of constant or varied widths T, B. They may be formed as integral parts of chamber walls 1101 , or attached by any suitable means. Widths T, B of features 1130 may be varied such that they are constant along their axial (e.g., top-bottom) runs or lengths L, or they may vary such that T = B, T < B, and/or T > B at any portions of their lengths L. An example of an advantage offered by use of configurations in which T < B is the improved filtration of smaller and larger particles. In given rotational conditions, the effect of Coriolis acceleration is higher for larger particles, thus pushing them towards the wedges 1130 on the side 1101. Since the wedges 1130 are narrower at the top and wider at the base, the Coriolis jetting will be downward rather than being upward, causing larger particles to be pushed down, which can assist separation.

[0053] Figures 6A and 6B are schematic diagrams showing further innovative features of embodiments of inertial separators 1 100, 1 150 in accordance with the invention, including namely variations of chamber bases 1102 in order to accomplish various desired results. As shown in Figure 6A, for example, bases 1 102 may be cupped or otherwise curved, so that they form walls of straight cross-section (Fig. 6A(1)), and/or of variably-sloped cross section (Fig. 6A(2)). Bases 1 102 having constant slope γ (relative to axis 1110) can be easier, and therefore less expensive, to fabricate; by controlling rotational speed, surface coating, and the slope γ, backsplash due to sample introduction, separation speeds and characteristics, etc., may be controlled. Variable slopes γ can be used to control flow transition from chamber bottom 1103 to walls 1 101 , and thereby sample retention times, etc.

[0054] As shown in Figure 6B, cupped chamber bases 1102 can be provided with (i.e., incorporate) grooves, channels, wedges, ridges, etc. 1130' in order to help guide sample particles 702 toward wall(s) 1101 and outlet(s) 1203 in various ways for various purposes. Such features 1130' may be of any desired configuration(s) - straight, curved (e.g., helical), etc, and may be of constant or varied widths T, B. They may be formed as integral parts of chamber walls 1 101 , or attached by any suitable means. For example, ridges or fins 1115 may be integrally molded or injected with, machined, or attached to bases 1102 and/or walls 1101. Widths T, B of features 1 130 may be varied such that they are constant along their axial (e.g., top-bottom) runs or lengths L, or they may vary such that T = B, T < B, and/or T > B at any portions of their lengths L. The incorporation of fins or ridges having holes 11 16 can be used to control back-splashing, sample mixing, and other processes during separation. They can also be used to improve the impartation of rotational motion to particle samples gather at the base 1 102 of the chamber, where, when relatively little friction is provided between the sample 702 and the interior surfaces of base 1 102 and/or sides 1101 , sample can tend to gather and avoid being processed.

[0055] Figures 7A - 7E are schematic diagrams showing innovative features of embodiments of centrifugal inlet channels or conduits 1 103 in accordance with the invention.

[0056] Figures 7A and 7B show embodiments of centrifugal chambers 1 150 which are adapted to rotate around inlets 1103, i.e., in which inlets 1103 do not rotate at all, or do not rotate at the same speed as the chamber body 1107. In such embodiments, it may be advantageous to use ball or other bearings between the rotating separator chamber body 1107 and the non-rotating/differently rotating inlet 1103. Solution(s) or other sample(s) 702 may be injected at entrance 1103e of the inlet, such that they flow into the chamber 1120 via inlet vent 1 103f; optionally such injection may be made while the chamber body 1107 is at rest, or moving. Among the several structural and processing advantages afforded by non-rotating inlets 1 103 is that separation within the inlet 1103 can be reduced or eliminated if/as desired.

[0057] As shown in Figures 7A, 7B, inlet channels 1 103 can be of a variety of configurations, depending upon their intended application(s), cost, and other factors, including materials etc. For example, they can be straight (Fig 7A), or curved. In the example shown in Figure 7B, separator inlet 1103 has a generally coiled, or helical, shape. Sample 702 injected at 722 flows downward and circumferentially, in a generally helical path, such that within the inlet channel 1103 the sample 722 is subjected to centrifugal forces, causing larger particles 703 to be pushed further toward the outside circumference of the flow path than the smaller particles 704. As in many tube flow conditions, parabolic laminar flow principles can cause smaller and larger particles 703, 704 to separate due to differences in flow speed, particle mass, and friction between particles, tube walls, and sample solution, etc.

[0058] Figures 7C - 7E show further embodiments of chamber inlets 103, and in particular embodiments adapted to rotate during separation processes. Such rotating inlet designs can be configured or rotation at the same speeds as their respective centrifugal chambers 1 107, or at different speeds. [0059] In the embodiment shown in Figure 7C, interior surface(s) 1113 of inlet channel 1103 comprise ridges, wedges, or grooves, etc. 1170. As the inlet channel 1103 rotates and sample solution 702 is introduced at 1103e, the sample flows downward in the direction of arrows 1103g and forms a generally parabolic flow profile as shown at 1103g. In addition, features 1170 induce a rotational spin to the flow motion. The rotation will result in a centrifugal force that can cause relatively larger particles 703 to move toward the outer circumferential regions of the inlet channel 1103, while smaller particles 704 remain closer to the center of the channel.

[0060] In the embodiment shown in Figure 7D, a single inlet channel 1103 is split into multiple parallel flow channels 1103', which are disposed in a pattern around axis 1 110 which may but in general not need be circular and/or centered on (e.g., concentrically arranged about) axis 1110. In such a configuration, rotation of the inlet channel 1103 can cause relatively larger particles 703 to move toward the outer circumferential regions of the inlet channels 1103', while smaller particles 704 tend to remain closer to the center of the channels. Parabolic flow effects can, as in other cases, be used to advantage.

[0061] As shown in Figure 7E, multi-channel inlet configurations 1 103 can comprise tubes disposed in multiple concentric rings. In the embodiment shown in Figure 7E, a set of relatively larger tubes 1103R is disposed in a generally circular pattern about axis 1 1 10, at a radius r1 , while a set of relatively smaller tubes 1103r is disposed at a radius r2, which in general is not equal to rl In the embodiment shown, two concentric sets of rings are shown. However, in various embodiments 3 or more sets of rings may be used.

[0062] As will be apparent to those skilled in the relevant arts, any or all of the features disclosed in Figures 7A- 7E may be used in combination. For example, interior surfaces of inlet tubes 1103 can be provided with ridges 1170, and/or as shown in Section D-D of Figure 7E; and set(s) of single straight tubes, helical or otherwise curved tubes, and/or multi-tube concentric and/or otherwise parallel structures (Figs 7D, 7E) may be used, e.g., in series.

[0063] Features of inertial separator outlets 1104, etc., are shown in Figures 8A - 8C. As shown in Figure 8A, rotation of chamber body 1107 can cause separated particles to flow upward and outward from the flow chamber 1 120, as shown by arrows 740, 744. Varying the speed of rotation of the chamber 1120 can enable the selection of which particles, relatively larger or smaller, are permitted to flow into interstage outlet 1203. Relatively smaller particles can be eluted by rotation at relatively lower speeds, and relatively larger particles by rotation at relatively higher speeds. Carrier solutions may be included as a part of sample 702 to assist in this process.

[0064] Alternatively, or in addition, as shown in Figure 8B one or more movable collecting channels 1193 may be used to draw off particles from any desired point(s) within the rotating chamber 1120. In the embodiment shown, for example, a tube 1203, 1 193 may be raised or lowered (M-M), translated laterally (L- L) and or rotated (N) in order to place a particle vent inlet 1 194 at any desired location within the chamber 1120, generally in close proximity to the chamber wall 1101. The use of suspension and/or solution buffers in the sample 702 can be used to help control this process. As will be understood by those skilled in the relevant arts, embodiments employing movable collecting channels 1193 enable the very precise elution of particles of specific size(s) according to any major or minor axis dimension(s), diameter(s), circumference(s), etc.

[0065] A further alternative or additional embodiment is shown in Figure 8C. In this embodiment channels 1130 and/or ridges 1130', which may be of differing widths, lengths, and configurations, may be used to guide particles of desired sizes toward suitably-sized holes 1133, 1 134, 1135 in walls 1 101 of chamber bodies 1 107. By using holes of varying size, arrangement(s), and number(s), for example with smaller holes 1 134 near the base 1 102 of the chamber and larger holes 1 135 further from the base, varying speeds, surface coatings, and solution buffers may be used to elute particles into a plurality of inter-stage channels 1203. Those skilled in the relevant arts who have made themselves familiar with this disclosure will understand that the sizes, numbers, locations, proximities, and arrangements of holes 1133, 1134, 1 135, channels 1130, and or ridges 1 130' may be used to control elution and separation of particles of a wide variety of sizes, masses, etc., depending upon the characteristics of the chamber 1 107, nature of sample 702, etc.

[0066] Figure 9 is a schematic diagram showing an example of mechanical support and installation for a centrifugal separation device 1100 in accordance with the disclosure. As shown in Figure 9, the body 1 107 of the separation device 1 100 can be attached to a motor or other device 1188 to drive rotation of the body, and cleaning system(s) 1182, including cleaning fluid and waste tanks, pumps, etc. can be provided for use as described below. The chamber 1107 is connected to the rotor 1188 at the bottom, or base 1102. The top part 1101 of the chamber (a.k.a the cap) can be fixedly or rotatably attached to the inlet 1103 for injecting sample solution 702. The inlet 1 103 can help to reduce or prevent vibration of the upper portions of the chamber 1107. Support for the chamber 1102 can alternatively, or in addition, be improved through the use of additional supports, such as ball- or other type bearings 1187 along the periphery of the body 1107. Additional bearings or supports can be provided at the top of the cap, for example bearings 1125 shown in Figure 7A.

[0067] As noted above, a plurality of centrifugal and/or other inertial separation components 1100 may be used, in parallel (e.g, multiple analysis pathways) and/or in series (for example, to improve refinement of particle separations).

[0068] Once a desired degree of particle separation has been accomplished by inertial means, separated portions 710 of sample 702 can be transferred to one or further stages of a hybrid separation system, for further separation through the application of principles based on other characteristics of the sample particles. For example, as shown in Figure 2, the separated particles may be passed to one or more electromagnetic field or flow (EFS) separators 1200 via interstage or interconnecting flow channels or conduits, 1203, for further-refined separation.

[0069] EFS separators 1200 suitable for use in implementing the invention can be of any form compatible with the purposes disclosed herein, and the specific applications to which they are to be put. For example, in many embodiments EFS separators comprising one or more arrays of capillaries disposed between charged plates, or other electrodes.

[0070] Figures 10A and 10B are schematic views of an EFS separator 1200 suitable for use as at least one stage of a separation component 100 of a system 1000 in accordance with the invention. In the embodiment shown, EFS separator 100, 1200 comprises a pair of flat sheet electrodes 1210a, 1210b and a capillary tube sample conduit or channel 1250 (which may also be referred to as a 'microchannel') disposed between the electrodes and configured to pass a sample 702 comprising particles carrying one or more distinct electric charges through an electromagnetic field established by passing static or variable current(s) through the electrodes. The sample 702, which has optionally been subjected to inertial separation process(es) by one or more inertial separation stages 1100, is introduced to the EFS separator 1200 via one or more interstage channels or conduits 1203 and passed into the conduit 1250. As the sample 702 passes through the capillary 1250 between the electrodes 1210a, 1210b, etc., electromagnetic field(s) set up by the currents are used to manipulate the charged particles within the sample and thereby separate them for detection and analysis by further stages 300 of the system 1000; the separated particles can be transferred to such later stages 300 via exit port(s) 1209.

[0071] Examples of charge schemes applicable to EFS separators in accordance with the invention comprise, for example, constant (DC) voltages, which can be applied to conduit walls and other electrodes; cyclical EFFF schemes, including DC or AC and variable & gradient voltage profiles, and other waveforms; and application of different electrode patterns to conduit walls or electrodes. In addition, or alternatively, neutral or ground electrodes may also be used in various embodiments.

[0072] Variations in control possibilities are enabled by the use of varied electrode and conduit configurations in EFS separators in accordance with the invention. For example, as shown in Figures 11A and 11 B, throughput can be increased by providing a plurality of conduits 1250a-x between the electrodes 1210a, 1210b. Throughput can further be increased by employing a plurality of EFS separators 1200, in parallel and/or series.

[0073] Another example is shown in Figures 12A and 12B. In the embodiment shown, EFS separator 1200 comprises a single capillary or conduit 1250, and two electrodes. One electrode 1210a' is positioned along axis 1237 or in another desired place within the capillary 1250; and cylindrical or other tubular capillary 1210b' encloses the capillary. As those skilled in the relevant arts will understand, once they have been made familiar with this disclosure, variations in relative sizes, material properties (including conductivity and capacitance), and shapes of electrodes 1210a', 1210b' can be used to tune, or optimize, the EFS separator 1200 for a variety of applications, and to control or influence construction costs.

[0074] As shown in Figure 13, capillaries and other conduits 1250 suitable for use in the invention can be modified for a wide variety of purposes. For example, all or parts of inner surfaces of the capillaries, i.e., surfaces exposed to analysis sample 702, can be coated with a variety of materials for specific applications. In the embodiment shown in Figure 13, for example, the inner surface of a conduit 1250 has been coated with electrically chargeable material, so that it can be induced to positive, negative, or neutral charge(s) (i.e., it can be made selectively into an anion, cation, or neutral surface/species), so that charged sample particles can be separated as desired. As noted, a variety of suitable charged or chargeable polymers are available, including, without limitation, polyamine, cationic cyclodextran, Poly(N,N-dimethylacrylamide) (PDMA), Polyethylenimine (PEI), etc. Suitable neutral polymers include, for example, alkylene-glycol poly (vinyl alcohol), copolymers of acrylamide, etc.

[0075] Figure 14 shows a further variation of an EFS separator 1200. The embodiment shown in Figure 14 combines features of the separators 1200 shown in Figures 11 and 12. In the embodiment shown, a plurality of conduits 1250a... 1250x are placed or otherwise disposed in a circular or other disposition (i.e., 'bundled') around an inner electrode 1210a', and encased by an outer electrode 1210b'. One or more inner electrodes 1210a' are disposed at the ends of the separator 1200, near the center (or at another desired position) of the conduit array 1250a-1250x. In addition to enabling increased throughput of any sample compositions, embodiments such as that shown in Figure 14 enable the simultaneous, or rapid, sequential separation of sample portions that have previously been separated according to inertial or other characteristics, using for example, one or more inertial separators 1100 as described herein.

[0076] Figure 15 shows a further variation of an EFS separator 1200. In the embodiment shown, two or more concentrically-disposed conduits 1250a, 1250b, etc., are disposed axially around an inner electrode 1210a', and inside an outer electrode 1210b. The aggregated conduit can be fabricated using, for example, a plurality of annularly layered porous glass capillaries or tubes, configured to act as micro- or nanofilters. Both the inner and outer electrodes 1210 may, like all electrodes described herein, be fabricated using materials and/or coatings that may be corrosion resistant, depending upon the application(s) to which they are to be put. For example, annularly-disposed porous Vycor glass (PVG) tubes may be used. Such configurations may be used with particular advantage in analyzing anionic particles, given the cationic nature of PVG.

[0077] Figure 16 shows a further variation of an EFS separator 1200. In the embodiment shown, no external capillary tube or Mylar spacer is used, rather electrodes 1210a' are formed into tubular, or other desired shape. Such electrodes and corresponding shapes can be molded or lithographically constructed to crate different channels with different shapes.

[0078] Figure 17 shows a further variation of an EFS separator 1200, in which filter(s) or filter layers 1270a-1270d of selected or otherwise variable mesh (pore sizes) are placed between electrodes 1210a, 1210b, to facilitate size- and charge- based separation. Filter(s) and/or filter layer(s) 1210a, 1210b, etc., may be used in many of the EFS separator embodiments described herein, as an additional separation means.

[0079] As previously described, hybrid separators, provided in distinct separation stages and operating on varying physical principles, may be used to advantage in separating particles in accordance with the invention. In addition, or in the alternative, hybrid separators may be provided in single-stage devices. In the above and other embodiments, for example, EFS separation processes may be conducted in combination with, and thereby assisted by, advantageous application(s) of other forms of energy, force, acceleration, and/or other physical phenomena, such as gravity, thermal phenomena, centrifugal acceleration, and additional electrical potentials/field generation processes. Individual separator stages 100, 1200 operating on such combined, or hybrid, principles may be described as single-stage hybrid compressors, which may be used singly or in combinations with any further desired forms of separators 100.

[0080] An example of such a single stage, hybrid separation system 1300 is shown in Figure 18. In the example shown in Figure 18, hybrid separation system combines an inertial separation component 1 100, in the form of a microchannel capillary 1252, 1250 having the shape of a helix or an Archimedean spiral, with an EFS component 1200 comprising electrodes 1210a, 1210b and capillary 1250, 1252. Inertial component 1 100 operates on both centrifugal principles and laminar and/or other flow principles, profiles, or regimes, as previously described. EFS component 1200 can further help to separate particles based on charge, as described above.

[0081] The use of curved microchannel(s) 1250, 1252 can result in improved separation efficiency, through the application of hydrodynamic-based microfluidic cell separation techniques. As a particular example, curved microchannels 1252 can provide separation efficiencies as high as 95%, compared to straight micro-channels 1250. This can, for example, arise through the use of curved microchannels 1250, 1252 in order to induce secondary flow patterns referred to as Dean vortices, which give rise to forces which may be used to separate particles of different sizes, masses, and/or densities. As previously noted, and intended throughout the remainder of the specifications, particle sizes in such contexts can include any or all of any major or minor axis dimension(s), diameter(s), circumference(s), etc.

[0082] The improvement in efficiency provided through the application of curved conduits may be even further increased through the concurrent application of EFS techniques. For example, the application of electric fields substantially perpendicular to the direction of flow in a curved microchannel 1250, as shown in Figure 18, can cause 2-dimensional separation based on both mass and charge. Centrifugal forces induced by the curved flow path can be used to separate charges, based on their masses, in radial directions, as shown in Section B-B of Figure 18. Differences in velocity of the carrier flow will separate particles, as shown in Section A-A of Figure 18. In addition, applications of steady-state or modulated electric fields can cause separation of particles in a desired direction on the basis of electric charge in a vertical direction.

[0083] Among the many advantages offered by hybrid EFS/inertial stages 1300, 1200, 1100 such as that shown in Figure 18 is that that they can be configured to operate without need of moving parts. This can help to reduce manufacturing and maintenance costs, as well as ease and efficiency of operation.

[0084] Some of the advantageous effects of the use of combined inertial/EFS hybrid separation systems are illustrated schematically in Figure 19. At (a), a representative chart of results of separation of a sample of microbes, using solely an inertial (e.g., centrifugal elutriation) separation system 1 100, 1 150 are shown. At 1801 a hypothetical, non-dimensional chart of sample mass flow vs. time is shown: this represents the relative amounts of sample mass 702 flowing into the separation subsystem 300 as a function of time, and is identical in all three cases (a) - (c). An illustrative schematic diagram of particle mixing in sample 702 for all three cases (a) - (c) is shown at 1847. In 1847 it is indicated that particles 703, 704 of sample 702 are mixed, with respect to size, mass, shape, and electrical charge, prior to any separation.

[0085] At 1802 in (a), it is shown (again hypothetically) that the centrifugal separator 1 100, 1150 has separated the sample 702 into portions centered around 3 peaks, representing a continuous mass spectrum dominated by three prominent distinct mass groupings. In other words, three distinct mass groupings are easily separable using the inertial system alone. This is further illustrated at 1848, where comparison with diagram 1847 suggests that the particles 703, 704 have been separated based on mass.

[0086] At 1803 in (b) of Figure 19, it is shown that an electric field-based separation system, such as an EFS separator 1200, analyzing an identical sample 702 based on particle charges has provided a charge grouping centered on four prominent distinct charges. Thus, the EFS separator has provided four easily- separable charge groupings. A schematic illustration of charges 703, 704 of a sample 702 separated based on charge is shown schematically at 1849.

[0087] In (c), however, it is shown at 1804 that a hybrid system has resolved an identical sample into seven distinct groupings, based on both mass, charge, and shape, in about the same amount of time required for either of the separate analyses. Thus, it may be seen that significant improvements in speed and quality of analysis are available, using systems in accordance with the disclosure. In practice, considerably greater improvements are readily achievable, using various aspects of the invention disclosed herein. It is also significant that such results are obtainable without finely tuning or optimizing individual separation stages. For example, a centrifugal stage 1 150 can be optimized to increase throughput while passing some unsorted smaller particles to an EFS stage 1200, as the EFS stage can be relied upon to provide further separation. [0088] Once a desired degree of particle separation has been accomplished, by single or hybrid separation means, separated portions 710 of a sample 702 can be transferred to one or more detection stages 200 (Figure 1 ), for example via interstage conduit(s) 2002.

[0089] Particle detectors 200 suitable for use in implementing the invention can be of any form(s) compatible with the purposes disclosed herein, and the specific applications to which embodiments of the invention are to be put. A wide variety of suitable technologies are known, and doubtless further examples will be developed in the future. For many applications, preferred embodiments of the invention use non-destructive technologies, such as those based on the scattering and detection of spatial distributions of light or other forms of particle excitation. The use of laser-based excitation technologies has been used with significant advantage, due to reliability, simplicity, controllability, and cost.

[0090] The use of non-destructive detection technologies offers many advantages, including for example the fact that biological particles may be preserved for further processing, for culturing to promote more exhaustive studies, etc.

[0091] For many applications, superior results can be obtained through the use of hybrid detection systems comprising the use of wide-spectrum identification techniques followed by narrow-band technologies. Such techniques enable, for example, an initial identification process targeting wide ranges of particles, followed by narrower spectrum detection processes to provide refined accuracies and identifications. The variety of exploratory sciences employed by the invention allow the attainment of accuracies of detection not previously possible.

[0092] Specific examples of non-destructive excitation / detection technologies suitable for use in implementing the invention include various forms of spectrometry, including for example Raman spectrometry (RS) for identification of bacteria, and multi-angle light scattering (MALS) techniques. MALS, for example, can be implemented in such ways as to produce unique light scattering patterns for each distinct microorganism, enabling resolutions down to about one tenth of the incident wavelength, or approximately 0.05 micrometers (μητι). Using MALS, any or all of the relative intensity, polarization, and phase of scattered light, considered as functions of scattering angle, can be used to identify a microorganism's unique dielectric composition.

[0093] As with other aspects of the invention, further advantage may be gained, in some applications, through the use of multiple particle throughput channels. This can be accomplished by any desired means, including for example parallel channels of similar detection stages, or through parallel channels based on different types of detection technologies.

[0094] As previously noted, the invention enables sorting, or partitioning, of particles based on a variety of characteristics or criteria. Particle detectors in accordance with the invention can, for example, be used to determine absolute or relative sizes of particles, counts of partitioned or unpartitioned groups of particles, and identities of various species of particles, and to partition particles according to such characteristics.

[0095] Figure 20 is a schematic diagram of a laser-based, non-destructive optical particle detection (sub)system 200. In the embodiment shown, (sub)system 200 comprises a plurality of particle detection, or excitation, devices 2102 in the form of lasers; one or more lenses 2200, adapted to focus and direct light generated by laser(s) 2102 toward any portion(s) of light detection chamber(s) 2300 containing particles of sample 702; one or more detection chambers, or conduits, 2300, which may be of any type suitable for transmitting light provided via lens(es) 2200 while adequately containing particles of sample 702, and fabricated using any materials suitable for that purpose; one or more light-scattering components 2400, adapted to condition light scattered by particles of sample 702 for detection by detection elements 2500; and one or more light detection elements 2500. In the embodiment shown, (sub)system 200 further comprises one or more optional detection lasers 21 12, which may be used to detect the sizes and/or locations of the perimeter of a cluster our other formation of sample 702, so that desired placement of the sample within the one or more detection chambers 2300, and/or determination of a sample size, may be accomplished.

[0096] Each of components 2102, 2112, 2200, 2300, 2400, 2500 may comprise any device(s) suitable for purposes disclosed herein. A wide variety of such devices are now available; doubtless others will be developed in future. Examples of currently-available excitation (e.g., light) sources include a variety of lasers, including for example semiconductor diode lasers, LEDs, solid state lasers, gas lasers, etc. Examples of currently-available suitable detectors include photodiodes, charge-coupled device (CCD) arrays, complementary metal-oxide- semiconductor (CMOS) arrays, photo-multiplier tube (PMT) detectors, avalanche photodiodes (APDs), etc.

[0097] The embodiment of Figure 20 is shown as a single-channel detection system. For many applications, multiple channel systems may be preferable. For example, in some configurations light-scattering components 2400 perform best when they are able to provide three-dimensional (3-D) data for sensing and interpretation by detector(s) 3500, processors 400, etc. Thus, for example, in some embodiments light-scattering components 2400 are configured to cause front, back, and/or side scattering of light.

[0098] In many embodiments, the use of multiple lasers 2102, 21 12, capable of generating light of differing wavelengths is preferred. The use of such arrays, or groups or banks, of lasers or other particle excitation sources can improve the amount and quality of scattering, which in turn can improve detection, identification, and other analysis of particles.

[0099] In embodiments of the invention employing lasers or other light sources as excitation devices 2102, the efficiency and accuracy of optical detectors 2500 in accordance with the invention can be improved through control of the polarization state(s) of input light sources 2102. Such polarization state(s) include any type of polarization suitable for a desired analysis, including for example circular, elliptical, planar, and/or linear polarization. The identification of structural details of small particles, for example, can in many circumstances be more accurately and efficiently accomplished by controlling polarization states than by the analysis of scattered light intensities alone.

[00100] In some embodiments, lenses 2200 and/or light-scattering elements 2400 comprise laminated or otherwise coated and/or polarized surfaces. For example, polarization optics, special coatings, etc., can be applied in order to improve focussing, filtering of selected wavelengths, etc. For example, beam focusing through the use of polarizing coating(s) and/or film lamination(s) can provide highly efficient and accurate results.

(00101] In various embodiments, detectors 2500 are optimized for detection of light of expected or otherwise desired wavelengths, in order, for example, to increase the speed, efficiency, and accuracy of identifications and other analyses. In many embodiments, multiple detectors 2500 are provided, such that they can effectively be combined to form large area detectors.

[00102] In various embodiments, selected combinations of laser(s) 2102 and lens(es) or other beam conditioning device(s) 2200 are used for a variety of purposes. For example, such combinations can be used to form optical beams suitable for manipulating particles of defined sizes, in order to improve identification, maneuvering, or other processing of the particles. Linearly and/and or circularly polarized light can be used to excite and/or otherwise manipulate particles for identification, maneuvering, etc. Phase- or wavelength-difference detection techniques may be used to identify and/or otherwise analyze particles.

[00103] Optical excitation may be induced using such combinations for, for example, creating characteristic scattering patterns, to be used in particle identification. Similarly, correlation of data signals generated by arrays or other pluralities of detectors 2500 may be used to identify and/or otherwise analyze particles.

[00104] In operation, a sample 702 provided, for example, via conduit(s) 2002 from separation (sub)system(s) 100, enters detection chamber 2300 in the direction shown by arrow 2301 , and is guided by the chamber to an analysis region as shown at 702 in Figure 19. In the analysis region the sample 702 is subjected to laser radiation generated by lasers 2102, 21 12, optionally conditioned by lens(es) 2200. Light scattered by impingement upon or other interaction (e.g., refraction or diffraction) with particles of sample 702 is further conditioned by light-scattering element(s) 2400, and detected by detection element(s) 2500. Corresponding signals generated by the detection element(s) 2500 may be interpreted by controller(s) 400 and used in identification and other processes.

[00105] In Figure 21 , a multi-channel (or multi-conduit) embodiment comprising a one-dimensional (1-D) array of detector elements 2500 is shown. As previously noted, the incorporation of multiple throughput channels 2300 can significantly reduce processing times. Alternatively, or in addition, the use of excitation devices 2102 of differing wavelengths, phases, polarization states, etc., and correspondingly- optimized detectors 2500 in conjunction with each of the throughput channels 2300 can be used to enable the simultaneous analysis of multiple samples comprising particles of differing characteristics. For example, the various modules, units, arrays, etc., of lasers 2102 shown in Figure 21 can be configured to provide any one or more desired wavelengths of light, and samples 702 can comprise portions of a single sample comprising statistically uniform mixtures of particles, or each of samples 702 can comprise particles previously sorted by mass, charge, and/or other properties, as described herein.

[00106] In Figure 22, a generalized schematic diagram of a multi-channel (or multi-conduit) embodiment comprising a plurality of throughput channels 2300 and a 2-D array of detector elements 2500 is shown. The 2-D array of detector elements 2500 can be coupled with arrays of correspondingly-configured lasers or excitation devices 2102 (not shown).

[00107] More specific embodiments of the generalized 2-D detector concept shown in Figure 22 are shown in Figures 23A - 23C. While some of the embodiments are shown as single channel (i.e., comprising a single flow throughput channel 2300), they can in general be used in parallel, or sequentially, in order to accomplish a variety of improvements, including increased analysis output rates, greater precision through the use of separate channels for separate classes of particles, etc. In general, each of the embodiments uses optical detection techniques to observe the interaction response between particles of interest with light excitation, through the detection and analysis of a variety of light characteristics, including but not limited to emitted and detected intensity(ies), phase(s), polarization state(s), 2D or 3D spatial scattering patterns, etc. Analysis of such characteristics can be accomplished through the application of automated signal processing algorithms, as described herein.

[00108] In the embodiment shown in Figure 23A, detector stage 200 comprises a detection chamber 2300 in the form of a capillary tube 2510 adapted for introduction of sample(s) 702 at an inlet 2002, and for allowing the sample(s) to pass a plurality of laser excitation devices 2102, 2102' and 2-D detector arrays 2500 in the form of one-sided or two-sided CCDs. Lenses 2200, mirrors 2525, and complete or partial anti-reflective (AR) coatings 2310 on capillary 2300 are provided to increase the efficiency, precision, and effectiveness of the detector stage 200.

[00109] As particles 703, 704, etc., pass along the length of the capillary 2300, they pass a plurality of laser excitation devices 2102, 2012' which are configured to provide light of differing wavelengths λι , λ ₂ , and polarization states Pt, P ₂ , and thereby to increase the amount and quality of scattering data collected by detectors 2500.

[00110] As may be seen in Figure 23A, mirrors 2525 can be used to effectively separate the capillary 1250 into a plurality of chambers 2300a, 2300b, 2300c, 2300d by reflecting light of specific wavelengths λι _, λ ₂ , polarization states P-i, P ₂ , etc. such that it is collected by specified detectors 2500, or portions thereof. Thus, for example, data acquired by detectors 2500 representing light of differing wavelengths may be decoupled ab initio, with corresponding improvements in the quality of data collected, and subsequent analysis processes.

[00111] Thus, for example, light emitted by laser 2102a may be seen to pass into region 707 of the capillary 1250, which corresponds to a detection chamber region 2300a. A portion of the light emitted by laser 1202a passes through the wall of capillary 1250 and strikes particles 703, 704, etc, of sample 702, and is reflected back out of the detection region 2300a and onto CCD sensor surfaces 2501 , 2502. A further portion of the light emitted by laser 1202a passes by the particles in region 2300a and strikes CCD detector 2503, either directly or after being reflected by mirror surface 2525a. Controller(s) 400 can record data representing the intensity, frequency, wavelength, time, and location of light beams captured by detectors 2501 , 2502, and 2503, and by comparing the data to known characteristics of the light emitted by laser(s) 2102a, determine reflection, diffraction, refraction, absorption, polarization state, and other characteristics associated with particles 703, 704 etc. present in the detection region 2300a at the time, and thereby deduce the length, shape, and other characteristics of the particles, and thereby identify them.

[00112] As will be appreciated by those skilled in the relevant arts, the use of AR coatings along the walls of capillary 1250 can decrease the amount of light reflected off the surfaces of the capillary without having first impinged upon or otherwise interacted with the particles 703, 704, etc., and thereby increase the signal-to-noise (SN) ratio, and quality, of data collected by sensors 2500, and thereby increase the efficiency, accuracy, reliability, and effectiveness of analysis processes according to the disclosure.

[00113] It may also be seen in Figure 23A that capillary 1250 / detection chamber 2300 is bent, or curved, rather than straight, as shown in Figures 20 - 22. The use of bent or curved capillaries/chambers (i.e., conduits) 1250, 2300 can facilitate the effective division of the conduits into multiple detection regions 2300a, etc., as described. In addition, because the flow path of particle samples 702 passing through the curved or bent conduits 1250, 2300 is longer than those of samples passing through straight conduits, the use of bent/curved conduits can facilitate the fabrication of compact detection stages 200, which in turn can be used to produce smaller, more readily portable or otherwise transportable systems 1000, which increases the variety of applications to which such systems may be put.

[00114] Figure 23B shows an alternative embodiment of a multi-wavelength, multi-polarization state detector stage 200. The embodiment shown in Figure 23b can, for example, enable improved efficiencies in the employment of two-sided CCD detectors 2500, thereby making it easier to implement multiple detection regions 2300a , 2300b, etc., and to provide compact system designs.

[00115] Figure 23C shows a further alternative embodiment of a multi- wavelength, multi-polarization state detector stage 200. The embodiment shown in Figure 23b comprises two flow paths 1250, 1250', which can be provided in parallel, so as to process separate samples 702, 702', or sequentially, so that by means of a connecting flow path (not shown) the same sample 702 can be subjected to repeated or distinct detection processes. In other words, excitation sources 2102c, 2102d can provide light at wavelengths λι, λ ₂ , and polarization states P-i, P2, respectively, like sources 2012a, 2012b, and thereby provide data similar to that provided by sources 2102a, 2102b, or they can provide excitation at separate wavelengths λ _3, λ ₄ , polarization states P ₃ , P ₄ , and thereby increase the diversity of data collected by sensors 2500.

[00116] Figures 24A - 24D illustrate further aspects of various embodiments of detector stages 200 in accordance with the invention. In particular, Figures 24A - 24D show embodiments of detector stages or devices 200 adapted to provide planar wave excitation to particle sample(s) 702 in detection chambers 2300. Planar excitation of particles can reduce or eliminate the need to maintain beam alignment and focus between the source 2012 of the excitation and the detection chamber 2300. Pluralities of excitation sources 2102 such as light-emitting diodes (LEDs) which emit light of different wavelengths can be combined in lateral dimensions to provide benefits such as further reduction or elimination of the need to focus light sources.

[00117] In the embodiment shown in Figure 24A, a light source 2102 such as a laser provides light in a wedged light guide plate 2106, which passes the light into a diffuser 2107 and thence into stacked prism films 2108 and plane polarizer(s) 21 10. Light passes out the polarizer(s) 21 10 along a planar wavefront, so that only substantially parallel light rays 2160 are passed through slit 2115 and between mirrors 2525. After passing between mirrors 2525, the substantially parallel light rays 2160 strike detection chamber 2300, which may be provided with anti-reflective coatings so as to increase signal-to-noise ratios as described above. As previously described, some of the light rays 2160 will be reflected by particles 703, 704, etc. of sample 702, while others will be diffracted or refracted around the particles. Light 2161 which is reflected from the particles will strike mirrors 2525, and reflected thereby, with a tendency to spread toward outer portions 2550 of detector(s) 2500. Diffracted or refracted light rays 2165 will tend to be captured by inner portions 2575 of the detector(s) 2500.

[00118] The use of selected slit widths 2115, curvatures for surfaces of mirrors 2525, and wavelengths and polarization states λ-ι, Ρ _Ί , etc., can enable very precise interpretation of signals generated by detector(s) 2500 representing times, locations, wavelengths, and polarization of light captured by the detector(s), and thereby improve identification and other analyses related to the particles 703, 704, etc. For example, the use of slits 2115 of approximately the same width as the diameter or width 231 1 of chamber 2300, and/or curved (e.g., concave or cone-shaped) reflective surfaces 2526 on mirror(s) 2525, can provide significant improvement in the diffusion of light across the breadth of detector(s) 2500, and thereby help to separate the sources and types 2161 , 2165, etc. of light captured by the detectors (sometimes called 'ray tracing'). [00119] A further means of helping to differentiate the sources or types 2161 , 2165 of light captured by detector(s) 2500 is the application of color-shifting films or coatings, or color converters 2526 to reflective surfaces of mirrors 2525, as shown for example in Figure 24B. Such films can, for example, lengthen or shorten the wavelengths λ of light reflected by the mirrors, and thereby help to distinguish light 2161 which has been reflected by particles 703, 704, etc, from light 2165 which has diffracted around the particles. Such information can be used to further distinguish, describe, and or identify the particles 703, 704. Examples of color converters suitable for use in implementing the invention include phosphors and/or quantum dots (QDs; i.e., semiconductor devices that tightly confine electrons or holes in all three spatial dimensions).

[00120] An alternative or additional means of helping to differentiate the sources or types 2161 , 2165 of light captured by detector(s) 2500 (ray tracing) is the use of additional detectors 2576 in place of mirrors 2525, to capture and interpret light 2161 reflected by particles 703, 704, etc., rather than reflect it back to regions 2550 of detector(s) 2500, as shown for example in Figure 24C.

[00121] An embodiment of a light source device 201 configured to improve detection and identification accuracy through the use of light, generated by a single source 2102, in multiple polarization states is shown in Figure 24D. In the embodiment shown in Figure 24D, device 201 comprises a liquid crystal layer 2742 that is controllable (e.g., tunable), through application of variable voltage(s), so as to enable selectable adjustment(s) of intensity and polarization state(s) of light 2160 emitted by the device 201. Such use of controllable liquid crystal layer(s) 2742 can allow the transmission light 2160 having multiple light polarization states P _n but the same wavelength λ and same optical path. Such embodiments allow in-situ signal differential for multiple polarization, and thereby enhance detection accuracy.

[00122] Figures 25A - 25C show further embodiments of detector stages 200 in accordance with various aspects of the invention.

[00123] Figure 25A shows a coaxial embodiment of a detector stage 200, adapted to provide for optical excitation of particles at multiple wavelengths, polarization states, etc. As shown in Figure 25A, the detector stage 200 comprises a plurality of excitation sources 2102a, 2012b, etc., arranged concentrically around the outside of an AR-coated detection chamber 2300 to form a source-detector ring 575, such that each of the sources 2012x is located diametrically across the chamber 2300 from a corresponding detector 2500x. Additional detector arrays 2500 can be located adjacent to the source-detector ring 575, to capture additional reflected, diffracted, and/or refracted light rays.

[00124] An alternative or additional embodiment of a coaxial detector stage 200 is shown in Figure 25B. In the embodiment shown in Figure 26, a plurality of angularly-displaced detectors 2576, 2577, 2578 is logically coupled to each excitation source 2102, so as to enable separate detectors to capture light rays 2161 , 2163, 2165 scattered by particles 702, 703, 704 etc in different ways. A plurality of sources 2102a, 2102b, etc., may be provided, and each may be separately coupled with the detectors 2576, 2577, 2578 according to various time and/or frequency division techniques, etc. For example, each of sources 2012a, 2012b, etc. may be activated at different times, so that light is captured by the detectors 2576, 2577, 2578 according to corresponding sources 2102a, etc., in time- sequential fashion; alternatively (or in addition) each of the sources 2102x and detectors can be coupled simultaneously but according to dispersed emission frequencies, so that scattering may be separately traced.

[00125] Coaxial arrangements such as that shown in Figure 25B can reduce dependencies between the rate of flow of sample(s) 702 and detection analyses, and therefore provide more reliable and consistent identifications, etc. Such arrangements can also be used to provide very compact design configurations, particularly as they can reduce or eliminate the need for light collimators and lenses.

[00126] A further alternative or additional co-axial detector embodiment is shown in Figure 25C. In the embodiment shown in Figure 25C, detector stage 200 comprises a wide spectrum light source 2102, which transmits light into a port 2177 and thence to a baffle 2178 adapted to diffuse the light 2160 through the detection chamber 2300. Spectrometer(s) 2195 at one or more detector port(s) 2180 can record color shifts caused by interaction(s) of the diffused light rays 2169 with the particles 702, 703, 704, etc., and separate them into spectrograms for subsequent analysis. Such arrangements can reduce or eliminate the need for multiple or variable excitation sources 2102, as described in connection with the various embodiments disclosed above. [00127] Specular reflection or component port 2190 shown in Figure 25C can enable capture and analysis of specular components 2167 of light reflected by particles 703, 704, etc. using detector(s) 2500 etc. Selective inclusion or exclusion of specular components (SCI or SCE) can improve the efficiency and quality of identifications and other analyses.

[00128] With particles of sample 702 suitably separated and identified, they may be passed, for example via conduits or channels 3002, to one or more collection stages or (sub)systems 300, for sorting or partitioning into temporary or indefinite containment, routing to other components of a system 1000 for further analysis processes, etc.

[00129] An example of a collection stage 300 in accordance with the invention is shown schematically in Figure 26A. In the embodiment shown, collection stage 300 comprises an inlet 3002 adapted for receiving sorted and/or analyzed sample(s) 702; one or more return loops, lines, or paths 390; one or more clamps 310; one or more particle detector system 315, comprising for example one or more laser(s) 321 and detector(s) 321 ; one or more carrier/buffer injection systems 370; and one or more containment ports or vents 340 and vent devices 344.

[00130] Collection of identified particles by (sub)systems 300 can be implemented using a variety of suitable technologies. For example, suitable devices can include cell-sorting and/or micro-fabricated collecting devices comprising mechanical or acoustical trap or drive means, and/or optical traps. Like separation stages 100, collection processes 300 can be based on the implementation of individual or hybrid technologies, and may be implemented in parallel and/or sequential multiples in order to increase processing speed and/or accuracy.

[00131] Return loop(s) 390 can be configured to return particle samples 702 to any desired one or more points in system 1000. This includes, for example, to one or more stages of a separation (sub)system 100 and/or detection (sub)system 200. Optionally, the point(s) in system 1000 to which samples 702 may be returned via loop(s) 390 may be manually and/or automatically selected, using suitably- configured input/output systems 406, 408, etc., of controller(s) 400. Particles 703, 704 and/or samples 702 can be routed into return loop(s) 390 using any or all of clamp device(s) 310 and diversion devices 340 such as those described below. [00132] Clamp(s) 310 comprise any means suitable for use in blocking and/or trapping sample(s) 702, or portions thereof, at desired points within the collection stage 300, and/or within any other component(s) of system(s) 1000, including any of stages 100, 200, etc. For example, mechanical (including acoustic), optical, and/or pneumatic means may be used. Opening, closing, and other functions of clamp(s) 310 may be conducted manually and/or automatically, through the use of controller(s) 400. They may be used, for example, to control timing of arrival of sample(s) 702 at desired points of the (sub)system 300, to return sample(s) 702 to desired portions of the system 1000 via return path(s) 390, etc.

[00133] Particle detector system(s) 315, comprising for example one or more laser(s) 321 and detector(s) 321 can be configured to detect the presence of particles at any one or more points with the collection stage 300, so that, for example, any fully- and/or semi-automated sorting, return, or other processes, such as sorting processes to be conducted at port(s) 340, may be prepared and timely conducted.

[00134] Carrier/buffer injection system(s) 370 can be configured to inject any desired or otherwise suitable buffer and/or carrier gas(ses) or other fluid(s) into a sample 702, for example to accompany the sample into container(s) 380. For example, inert or other fluid(s) may be used to preserve samples, prevent corrosion or contamination of container(s) 380, protect laboratory personnel, etc.

[00135] Containment port or vent device(s) 344 can comprise any suitable or otherwise desired means, including for example acoustic, vacuum and/or laser- manipulation systems, configured for ushering sample(s) 702 into vials, tubes, or other containers, for storage, inculturation, further analysis, etc.

[00136] A schematic diagram of acoustic wave device(s) 344, 346 suitable for use in collection system(s) 300 in accordance with the invention is shown in Figure 26B. In the embodiment shown in Figure 26B, a plurality of acoustic devices 346, such as ultrasound devices, are adapted to provide focused acoustic waves directed at ports 340. As sample(s) 702 and/or separated or partitioned particles 703, 704 enter the vicinity of vent port(s) 340, acoustic waves of suitable frequencies and intensities may be directed at the port, and thereby push the particles into container(s) 380 acoustically. [00137] A schematic diagram of vent device(s) 344, 349 based on differential pressures, suitable for use in collection system(s) 300 in accordance with the invention, is shown in Figure 26C. In the embodiment shown in Figure 26C, a plurality of differential pressure devices, such as micro-pumps 349 adapted to create suction(s) sufficient to draw particles 703, 704, etc., into containers 380, are placed at ports 340. As particles 702 approach a port adjacent desired container(s), corresponding pump(s) 349 may be activated, causing the particles to be drawn into the container(s).

[00138] A schematic diagram of optical vent device(s) 344, 351 suitable for use in collection system(s) 300 in accordance with the invention is shown in Figure 26D. In the embodiment shown in Figure 26D, which is a schematic section of channel or conduit 345 taken at section 26D-26D of Figure 26A, comprises laser or other optical excitation source(s) 320 and rotatable or otherwise deflectable mirror(s), waveguide(s), or other beam deflector(s) 357, positioned proximate to vent port(s) 340. As desired particle(s) 703, 704 pass along the conduit 345 proximate the port(s) 340, excitation source(s) 320 can be activated, to cross through the conduit 345 on a side of particle(s) 703, 704 distal from the port(s) 340 (i.e., 'behind' the particles), as shown at 359a. Beam deflector(s) 357 may be activated so as to cause the light beam 359 to shift to a direction 359b, and in doing so to impel the particle(s) 703, 704 to move into container(s) 380. Control of beam deflector(s) 357 can be accomplished by any suitable means, which may for example include suitably-configured microelectromechanical system(s) (MEMS) actuators coupled to mirror(s), waveguide(s), or other beam deflector(s) 357, causing the beam deflector(s) to rotate from a position 1 to a position 2, as shown in Figure 26B. A variety of beam directing or deflecting devices may be used, including for example suitably-configured lenses, polarizers, etc.

[00139] Further embodiments of improvements in particle sorting and containment systems 300, in the form of MEMS routing devices 385, are shown in Figures 27A and 27B. Such devices may be referred to as particle and/or sample flow control devices. It should be noted that the embodiments of the routing device(s) 385 shown are suitable for use not only in implementation of sorting and collection systems 300, but may be used generally in any suitable locations in systems 1000, in order to route particles 703, 704, etc., and/or sample(s) 702 to desired stages or components 100, 200, 300, etc., including for example through the use of return loop(s) 370.

[00140] As a particular example, routing device(s) 385 can comprise microvalves or switches comprising rotatable or otherwise re-directable conduit sections 386, controlled by MEMS devices 389. As shown for example in Figure 27A, re-directable sections 386 can selectively connect conduits 345, 2300, 1250, etc., with pluralities of alternate downstream paths or conduits 381a, 381 b, etc., as desired, in order to route particle(s) or particle group(s) 702, 703, 704, etc., to desired downstream system(s), containers 380, etc.

[00141] An important aspect of the use of routing device(s) 385 in accordance with the invention is that, as shown in Figure 26B, they may be employed, in series or and in parallel, in any desired combination(s), with any desired components, including for example, any or all of stages or subsystems 100, 200, 300, and/or returns 390 between them, in order to facilitate any desired routing of sorted or unsorted, identified, or unidentified particles or samples 702, 703, 704, etc.

[00142] Container(s) 380 may be of any type suitable for use in specific applications, for example glass, metal, ceramic, or composite tubes, vials, etc.

[00143] As in all (sub)systems 100, 200, 300 etc. in this disclosure, sample(s) 702 may be moved to from point to point, to desired position(s) within the collection stage 300, through any suitable means, including for example differential pressures, electrical, optical and/or mechanical (including acoustic) forces, etc.

[00144] As noted above, cleaning and other conditioning of system(s) 1000 and their sub(systems) 100, 200, 300, and components thereof, can be an important part of their maintenance and operation. Cleaning and other conditioning processes can be important both for ensuring the purity of samples to be analyzed and for ensuring the maintainability of the systems, and the health of humans who use them. Moreover, such processes can be conducted in a wide variety of manners. In various aspects and embodiments the invention provides a number of innovative processes for cleaning and other conditioning of particle analysis systems.

[00145] In general, cleaning and other conditioning processes can be accomplished wholly or partially through the introduction of sterilizing, biocidal, buffering, anti-corrosion, and/or other agents to portions of systems 1000, (sub)systems 100, 200, 300, etc., and/or components of any thereof, or by replacement thereof.

[00146] For example, interior surfaces of components of a system 1000, including particularly those surfaces that may be expected to come into contact with samples 702 and/or particles 703, 704, etc., can, for many applications, be coated with, exposed to, or fabricated using biocompatible and/or corrosion resistant materials such as epoxy novolac or metallic alloys. As will be understood by those skilled in the relevant arts, the selection of appropriate materials will depend upon the type(s) of particles to be processed, the characteristic(s) of components to be treated, including the materials of which they are made, and in many cases the purpose(s) of the analyses to which the systems and components are to be put.

[00147] Components such as centrifugal separation chambers 1107, 1120, detection chambers 2300, capillaries 1250 and other conduits or channels 2300, 345, etc., and other portions of systems 1000 exposed to samples and/or particles during processing, can be configured with efficient cleaning and sterilization in mind. As shown in Figure 9, for example, a drain 1160 can be provided at the bottom 1103 or other appropriate portion of a chamber body 1 107 of a centrifugal separator stage 100, 1100. During an example cleaning process, the chamber body 1 107 can be held stationary while suitable portions of the chamber 1120 are filled with cleaning agent. The chamber can be made to rotate at relatively low rates, typically much slower RPMs than are used for analysis. This can, for example, induce centrifugal effects in the cleaning agent, and thereby reduce the amount of the cleaning agent required for sterilizing/cleaning desired surfaces of the chamber. Once the cleaning process is finished, the cleaning agent can be dumped into a holding tank at 1182 in Figure 9.

[00148] Alternatively, components such as separation chambers 1107, separation chambers 1107, 1 120, detection chambers 2300, capillaries 1250 and other conduits or channels 2300, 345, can be provided in removable form, and either removed for cleaning, or simply replaced, following or in advance of, separation, detection, sorting, containment, and other processes. The use of disposable / replaceable components can be advantageous in terms of efficiency and effectiveness of analysis processing, sanitation / health concerns, and cost effectiveness of systems and components in accordance with the invention. [00149] Thus, in various aspects and embodiments, the invention provides systems, processes, and corresponding computer program instruction sets for self- contained cleaning and conditioning of individual stages or (sub)systems 100, 200, 300.

[00150] In various further aspects and embodiments, the invention provides systems, processes, and corresponding computer program instruction sets for self- contained cleaning and conditioning of complete systems 1000, or at least the entire sample processing chain, consisting of all conduits, chambers, etc., through which particles pass, or in which they are otherwise exposed during separation, detection, and/or collection/sorting, or containment.

[00151] To ensure the accuracy and safety of various analyses, any cleaning processes in accordance with the invention can be incorporated as automatic, semiautomatic, and optionally over-ridable parts of system or stage power-up and/or reset processes. Such options can, of course, be provided, managed, and modified through the proper programming and operation of controller(s) 400. In various embodiments, such power-up sequences can proceed in accordance with coded instruction sets applying algorithms adapted to maximize surface cleaning treatments, based on centrifuge rotation rates, system and/or subsystem flow rates, electrical potentials applied in EFS separation systems, etc.

[00152] In accordance with one embodiment of a power-up sequence in accordance with the invention, an operator of a controller 400 is enabled to choose between one, some, or all of components 1000, 100, 200, 300 for cleaning, e.g., to remove contamination, prior to particle processing operations. Such start-up processes can start with treating contact surfaces (e.g., surfaces of particle pathways and containment compartments) with one or more treatment agents, by injecting the agents through inlets/ports 1002, 2002, 3002, etc., and allowing them to pass through some or all of the particle processing chain or pathway. As one specific example, an inside surface of a capillary tube 1250, 345 or conduit 1002, 2002, 3002, etc., can be coated with buffering agent to reduce particle fiction. The efficiency and/or effectiveness of such cleaning processes can further be improved by, for example, activating moving parts of the system 1000, such as separation chambers 1107 and/or mechanical clamps, gates or valves; and/or by applying electrical fields in EFFS/detection stage(s) 200 etc. [00153] As previously mentioned, one or more controllers 400 can be used to control operation of any portion(s) of systems 1000, in order to initiate, control, oversee, and record the results of any desired processes, including any or all of the separation, detection, sorting, containment, and data reduction or analysis processes described or suggested above. Such controllers may be provided with any number(s) of local and/or remote (e.g., cloud-based) data storage systems 404, in order to store coded machine readable instruction sets, processing and/or analysis data, etc. Such instruction sets, analysis data, etc., may be used in real-time or delayed analyses, archived for further research, data reduction and analysis, etc.

[00154] Collection, retention, analysis, and other processing of data collected by systems 1000 in accordance with the invention, and particularly data collected by sensors 2500, etc., of detection stages or (sub)systems 200, is among the important advantages offered by systems 1000 in accordance with the invention, and components thereof, including any or all of (sub)systems 100, 200, 300, 400. For example, data representing wavelength, phase, polarization state, time, and location of light, generated using signals provided by sensors 2500; data representing pressures generated by gauges along flow communication lines 1002, 2002, 3002, 1250, 2300, 345, etc.; switch, clamp, and/or gate states provided by MEMS and other devices 310, 389, etc.; can all be stored by controller(s) 400 in memory(ies) 404 and processed, in real time or later, to interpret and otherwise analyze particle identities, system operational functions and performance, etc.

[00155] Thus, for example, the invention provides, in various aspects and embodiments, systems, devices, and machine-readable instruction sets configured to cause one or more automatic data processors to acquire, store, analyze, and otherwise process particle identification and other data generated by separation, detection, sorting, and containment components described or suggested, including without limitation the generation of data sets comprising data useful for classification of particles and/or data representing characteristic responses of particles to optical and other detection system components.

[00156] Similarly, such controllers 400 and data stored in memory(s) 404 can be used to generate suitable command signals and thereby to control operations of devices such as gates 389, clamps 310, micropumps 349, acoustic devices 344, 346, etc., and thereby control the flow of sample(s) 702 and particles 703, 704, etc., through system(s) 1000 and their various subsystems, stages, and components, and data collection and analysis processes as described herein.

[00157] While the disclosure has been provided and illustrated in connection with a variety of specific, presently-preferred embodiments, many variations and modifications may be made without departing from the spirit and scope of the invention(s) disclosed herein. The disclosure and invention(s) are therefore not to be limited to the exact components or details of methodology or construction set forth above. Except to the extent necessary or inherent in the processes themselves, no particular order to steps or stages of methods or processes described in this disclosure, including the Figures, is intended or implied. In many cases the order of process steps may be varied without changing the purpose, effect, or import of the methods described. The scope of the invention is to be defined solely by the appended claims, giving due consideration to the doctrine of equivalents and related doctrines.

Selected features from one or more of the above-described embodiments may be combined to create alternative embodiments not explicitly described, features suitable for such combinations being readily apparent to persons skilled in the art. The subject matter described herein in the recited claims intends to cover and embrace all suitable changes in technology.