BACKGROUND TO THE INVENTION Biological tissue, for example plant or animal tissue, is analysed for a vast range of purposes using various tissue processing techniques. For example, tissue may be required to undergo DNA extraction, forensic analysis or the like. A multi-stage process is usually required, one stage of which is tissue disintegration. Tissue needs to be broken down or disintegrated to release the cells for subsequent processing and analysis stages. This is usually achieved by placing samples of the tissue in a ninety six-well plate or similar, and manually securing the well plate in a clamp of a shaking device, which vigorously vibrates the tissue (in the presence of some form of grinding beads) to perform disintegration.

The various stages involved in processing tissue for analysis are labour intensive if carried out manually. Robotic arms and automated analysis equipment have been designed for automating some tissue processing tasks. However, at some point, manual intervention is required to place the tissue samples in a disintegrator, and then retrieve them once tissue disintegration is complete.

SUMMARY OF THE INVENTION It is an object of the invention to provide a disintegrator that can automatically receive and disintegrate tissue samples and/or which at least provides the public with a useful choice. Preferably such a disintegrator can be integrated into an automated sample processing system to remove the need for manual intervention to carry out tissue disintegration.

In accordance with a first aspect of the present invention, there is provided an apparatus for tissue disintegration including: a holder for retaining a sample or samples of tissue; a shaking mechanism for shaking the holder; and a controller adapted to receive one or more trigger signals for operating the holder to secure and release the sample (s), and for operating the shaking mechanism.

The holder may be configured to receive a sample receptacle containing the sample (s).

The holder comprises a clamp to clamp the sample receptacle. Preferably, the clamp includes a base part for supporting the sample receptacle and a clamping member to clamp the sample receptacle against the base part.

The apparatus preferably includes an actuator to move the clamp between a released position in which the sample receptacle is released and a clamped position in which the sample receptacle is secured in the holder, which actuator may be a pneumatic ram.

The controller may be configured to operate the actuator in response to the trigger signal (s).

The shaking mechanism advantageously includes a motor with an offset cam operatively connected to the motor and configured such that rotational movement of the offset cam shakes the holder. The controller may include a speed controller for the motor, and may be configured to operate the speed controller in response to the trigger signal (s).

The apparatus is preferably used to disintegrate tissue.

In accordance with a second aspect of the present invention, there is provided a tissue sample processing system including: a disintegrator with a holder for retaining a sample or samples; a holder shaking mechanism and a controller for operating the holder shaking mechanism; a robotic device for placing the sample (s) in and retrieving the sample (s) from the holder; and a trigger source for triggering the controller to secure the sample (s) when placed in the holder by the robotic device, operate the shaking device, and release the sample (s) for retrieval by the robotic device.

The holder is preferably configured to receive a sample receptacle. The holder may comprise a clamp to clamp the sample receptacle. Preferably, the clamp includes a base part for supporting the sample receptacle and a clamping member to clamp the sample receptacle against the base part.

The system may include an actuator to move the clamp between a released position in which the sample receptacle is released and a clamped position in which the sample receptacle is secured in the holder, which actuator may be a pneumatic ram.

The controller may be configured to operate the actuator.

In one embodiment, the shaking mechanism includes a motor with an offset cam operatively connected to the motor and configured such that rotational movement of the offset cam shakes the holder. The controller preferably includes a speed controller for the motor.

Preferably, the trigger source is configured to send a process initialisation signal, and the controller is configured to secure the sample (s) when placed in the holder by the robotic device, operate the shaking device, and release the disintegrated sample (s) following disintegration for retrieval by the robotic device based on time delays.

Alternatively, the controller may be configured to secure the sample (s) when placed in the holder by the robotic device in response to a trigger, configured to operate the shaking device in response to a further trigger, and configured to release the sample (s) for retrieval by the robotic device.

Advantageously, the robotic device includes at least one robotic arm. The system preferably includes a controller to operate the robotic device. The controller to operate the robotic device may communicate with and trigger the disintegrator controller.

The system preferably includes a liquid processing system for carrying out analysis and/or further processing of the sample (s). The system preferably includes a controller to control the liquid processing system. The liquid processing system controller suitably communicates with and triggers the disintegrator controller and/or the robotic device controller.

At least the liquid processing system and robotic device may be controlled by one controller.

In one embodiment, the robotic device is configured to retrieve the sample (s) from the holder and deliver the sample (s) to part of the liquid processing system for analysis and/or further processing. The robotic device is suitably configured to retrieve the sample (s) from the liquid processing system following the analysis and/or further processing.

The robotic device may be part of the liquid processing system.

The robotic device preferably includes a robotic arm configured to interchangeably carry a number of items.

The robotic device may include two robotic arms, at least one of which is configured to carry a gripping device for gripping tissue sample receptacles.

The other robotic arm may be configured to carry at least one pipetting device for extracting liquid from the sample (s).

The processing system preferably further includes a cool storage unit for storing the tissue sample (s) prior to disintegration and/or sample (s) following analysis and/or further processing. The robotic device may be configured to transfer the sample (s) from the cool storage unit to the disintegrator. The robotic device may be further configured to retrieve the sample (s) from the liquid processing system following the analysis and/or further processing, and to deliver the sample (s) back to the cool storage unit.

The liquid processing system may be configured to extract one or more components from the sample (s). The component could be a molecule such as DNA or a protein, a compound, or an element for example. The system has application for a wide range of samples of organic or inorganic materials, such as biological tissue, paint chips, etc.

The system has application for metabolomics, forensic, medicine, etc.

The liquid processing system is preferably configured to extract DNA from the sample (s).

The system is preferably configured to deliver sample (s) to the holder of a disintegrator, 'operate the disintegrator, retrieve sample (s) from the holder and deliver the sample (s) to a liquid handler for further processing and/or analysis, further process and/or analyse the sample (s), retrieve the sample (s) from the liquid handler, and deliver the sample (s) to a cool storage unit, automatically.

The system is preferably used to disintegrate and process sample (s) of tissue.

In accordance with a third aspect of the present invention, there is provided a method of processing sample (s) including: placing the sample (s) in a holder of a disintegrator using a robotic device ; sending one or more trigger signals to a controller to operate the holder to secure the sample (s), operate a shaking mechanism to shake the holder, and operate the holder to release the sample (s); and removing the sample (s) from the holder using the robotic device.

The tissue sample (s) is/are suitably placed in a receptacle that is adapted to be placed in the disintegrator holder. Preferably, the sample (s) is/are placed in at least one well of a multiple-well plate, which is adapted to be placed in the disintegrator holder.

The holder may comprise a clamp to clamp the receptacle.

The clamp preferably includes a base part for supporting the receptacle, and a clamping member to clamp the receptacle against the base part, and the method includes clamping the receptacle between the base part and the clamping member prior to shaking the holder.

The shaking mechanism may include a motor with an offset cam operatively connected to the motor and configured such that rotational movement of the offset cam shakes the holder, and the method suitably includes operating the motor to shake the holder.

The method preferably includes initially extracting the sample (s) from a cooled or otherwise storage position and delivering it/them to the disintegrator.

The method preferably further includes delivering sample (s) from the disintegrator to part of a liquid processing system for carrying out analysis and/or further processing of the sample (s). The method preferably includes extracting a component from the sample (s). More preferably, the method includes extracting DNA from the sample (s).

The method may include retrieving the sample (s) from the liquid processing system following the analysis and/or further processing. The method suitably includes delivering the sample (s) from the liquid processing system to a cool storage unit.

Preferably, delivery of the sample (s) to the holder of the disintegrator, operation of the disintegrator, retrieval of the sample (s) from the holder and delivery to a liquid handler for further processing and/or analysis, further processing and/or analysis of the sample (s), retrieval of the sample (s) from the liquid handler, and delivery of the sample (s) to the cool storage unit, is automated.

Preferably, the sample (s) is/are tissue.

BRIEF DESCRIPTION OF THE DRAWINGS Preferred embodiments of the invention will be described with reference to the following drawings, in which: Figure 1 shows a perspective view of the holding and shaking mechanism of a preferred tissue disintegrator; Figure 2 shows a suitable controller for the disintegrator; Figure 3 shows one possible form of a holder in more detail; Figure 4 shows the shaker crank shaft and asymmetric cam of the holding and shaking mechanism in this particular embodiment; Figure 5 shows a plan view of the holder and shaking mechanism; Figure 6 shows a block diagram of an embodiment of a DNA extraction system; Figure 7 shows one integrated tissue processing system incorporating the tissue disintegrator; Figure 8 shows a flow diagram of one possible component extraction process (in this case DNA) ; Figure 9 shows a plan view of an alternative embodiment tissue processing system for DNA extraction utilising an alternative preferred tissue disintegrator; Figure 10 shows a perspective view of part of the tissue processing system of Figure 9; Figure 11 shows a detailed front elevation view of the tissue disintegrator of the system of Figure 9 ; and Figure 12 shows an end elevation view of the system of Figure 9.

DETAILED DESCRIPTION OF THE INVENTION Figures 1 to 6 show a preferred embodiment of the disintegrator 1 for disintegrating tissue according to the invention, adapted for integration into an automated sample processing system. The disintegrator 1 includes a holder 10 (shown in detail in Figure 3) in the form of a clamp with a top clamp plate 11 and a base part in the form of a plate 12. The base plate is fixed to a shaking arm 18 that is pivotally hinged 19 to a distal end of the apparatus 1. The clamp plate 11 is pivotally hinged 13 to a clamping arm 14 that in turn is pivotally hinged 15 to a spacer arm 16 extending from the shaking arm 18.

The clamp plate 11 is adapted to clamp on a tissue receptacle 20, for example a multiple-well plate (such as a ninety six-well plate) or similar, placed on the base plate 12 and thereby secure the receptacle 20 in the holder 10. The clamp plate 11 is operated by an actuator such as a pneumatic ram 21 connected via a pivot link 17 to the clamping arm 14. Alternatively, the actuator could be a hydraulic ram or an electric solenoid actuator or similar. To clamp the receptacle 20 in place, the ram 21 extends upwards and rotates the clamping 14 arm about the pivot 15, bringing the clamp plate 11 down 22 on to the receptacle 20. To release, the ram 21 retracts, pulling the clamping arm 14 down, and thus the clamping plate 11 up 23. The clamping plate covers the well plate during shaking to prevent material escaping. Additionally, the well plate could include foil or plastic covers or similar, which could be pierced or removed for subsequent processing steps. The pneumatic ram 21 is controlled via air lines 24 that are connected to a compressed air source via solenoid valves. The holder 10 is vibrated by way of a crank shaft 25 (portions of which are visible in Figures 1,3 and 4) that is pivotally attached 26 at one end to the spacer arm 16 of the holder 10, and pivotally attached 27 at the other end to an offset cam 28. The offset cam is rotated by a drive shaft 38 that is driven by a dc motor 29, which pushes the crank shaft 25 up and down to shake the holder 10.

Operation of the holder 10 and vibration mechanism is controlled by the controller 30 such as that shown in Figure 2. It includes a speed controller 31 for the dc motor 29, power supply 32, and a microprocessor or microcontroller (such as a BX24) and associated control circuitry 34. It also includes an RS232 port 35, and a signal line 33 for receiving external signals from other equipment in a biological or chemical processing/analysis system, along with power connections 36 for the motor and controller and manual start and stop buttons 37. The interface 35 for external signals enables the tissue disintegrator 1 to be integrated with an automated system such that it can be controlled to operate at required times. The controller 30 receives one or more external trigger signals from a controller (such as a microprocessor, computer or microcontroller) or other equipment in the overall system, which initiate and/or control (in conjunction with the microprocessor 34) operation of the holder 10 and vibration mechanism. This for example, might be a sole signal which triggers the microprocessor to clamp the holder 10, start vibration, cease vibration, and release the clamp, based on time delays. Alternatively, several external triggers may initiate each of the steps mentioned above.

Figure 6 shows a block diagram of one possible application of the disintegrator 1 in an automated DNA extraction system 40, and Figure 7 shows one possible physical arrangement of an integrated sample processing system. The system includes a freezer 41 containing tissue samples in one or more multiple-well plates, a liquid handler (for example, a BIOMEK 2000) 42 for extracting DNA, and a robotic arm (for example, a ZYMATE XP) 43 for transferring the multiple-well plate between the freezer 41, tissue disintegrator 1, and liquid handler 42. A PC computer or similar 43 controls the liquid handler via a RS232 serial link, and interacts with a PC and controller 44 which overseas operation of the robotic arm 45.

As shown in the flow diagram in Figure 8, during operation, the freezer 41 is opened by way of a pneumatic ram or similar. The robotic arm 45 which rotates about its central pedestal and extends in the directions of arrows U and V respectively, grasps a multiple- well plate 20 containing tissue from the freezer 41 and carries it to and places it in the holder 10 of the disintegrator. The freezer is closed. The disintegrator clamps and secures the multiple-well plate 20 and then shakes it for the required duration. Upon completion, the clamp is released, and the robotic arm grasps the multiple-well plate and places it in the liquid handler 42 as indicated at 48. Once the liquid handler has completed DNA extraction, the freezer is opened and the robotic arm 45 grasps the multiple-well plate and places it inside for storage. This describes the process for one multiple-well plate, however it will be appreciated that a plurality of well plates may be processed simultaneously at different stages in the system.

The interaction of the disintegrator 1 with the remaining DNA extraction system 40 and process will be generally described. An initiation string is sent from one of the external devices (such as the robotic arm, PC, microcontroller or the like) to begin the disintegrator function. This string is sent once the multiple-well plate 20 is placed in the base plate 12 by the robotic arm. The tissue disintegrator microcontroller 34 receives (via the RS232 link) and checks the validity of the string and returns an acknowledgement if the string contains correct values specifying vibration duration and intensity values. The microcontroller 34 then activates the solenoid valves to extend the ram 21 and clamp the multiple-well plate 20 in position. Once done, the microcontroller 34 triggers the speed controller 31 to ramp up the speed of the shaft 38 until a speed is reached that provides the specified shaking intensity of the holder 10 via the cam/crankshaft 25 arrangement. After the specified shaking time has elapsed, the microcontroller 34 instructs the speed controller 31 to ramp down the dc motor 29 speed until it stops in the desired position. Some type of suitable feedback control or locking mechanism can be used to bring the base plate into the required position. A brake or locking member may be configured to engage the shaft 38 or offset cam 28 to quickly stop the holder 10 in the desired position. The microcontroller 34 then releases the valves to retract the ram 21 thus releasing the clamp plate 11 from the multiple-well plate 20. The microcontroller 34 then sends a terminating string to the originating controller, which triggers the robotic arm to remove the multiple-well plate 20 and enables the system 40 to continue with the remainder of the DNA extraction process.

The particular configuration shown in Figure 6 and its interaction with the tissue disintegrator 1 will be described in more detail. The B724 microcontroller 34 receives the initiation string from the liquid handler 42, operates the freezer 41 opening mechanism and auto latches (required to anchor resources to the BIOMEK 2000 work surface), in addition to operate the holder 10 and vibration mechanism, although other control configurations are possible. For example, the microcontroller 34 may only control the disintegrator functions. Due to the fact that the liquid handler 42 performs the majority of the DNA extraction process, has five communication ports, and has the most advance programming capabilities, it is used as the core communicator/controller of the system. It will be appreciated that a separate controller could be used for overall control/coordination of the system. The control could be via a PC card or the like.

The microcontroller 34 runs in an eternal loop until data arrives in the input buffer from the liquid handler 42. Upon arrival, one byte is removed and stored as a variable which is used in a switch/case statement to determine which device the following byte is addressed to. Where the variable = 1, the subsequent byte of data allows the execution of freezer specific code. The second byte is again used in a switch/case statement to determine whether the freezer door is opened or closed. The relay required to open or close air flow to the pneumatic cylinders is activated/deactivated by setting pin seven of the microcontroller 34 to five or zero volts respectively. A switch in parallel with the relay allows manual opening/closing of the freezer 41. Upon completion, an acknowledge string is returned to the liquid handler 42, and the input buffer cleared.

If the variable = 2, then the following two bytes are used by code specific for the disintegrator 1. The first byte determines the required intensity (1-16), while the second sets the desired length of disintegration (5-255 seconds). Having the relevant settings, an initiation request is sent to the disintegrator 1. An acknowledge is received from the disintegrator when the initialisation string is accepted, and also when the process of disintegration is complete. The microcontroller 34 then returns an acknowledge to the liquid handler 42 and clears the input buffer.

If the variable = 3, the following byte is used in the auto latch specific code. Auto latches are required to secure racks of the disposable pipette tips required by the liquid handler for liquid transfers. The second byte of the control signal supplies the auto latch number (1-4) and an open/close digit. When the appropriate action is taken by setting pins 13-16 on the microcontroller 34, a corresponding acknowledge is sent to the liquid handler 42 and the input buffer is cleared.

It will be appreciated that the communication channels and configurations shown in Figure 6 are exemplary only and other configurations are possible. Further, the system shown in Figures 6 and 7 is exemplary of one of many automated processing systems that could utilise the disintegrator 1. Further examples include metabolomic, chemical, biological, medical, and forensic analysis systems. It should be appreciated that other control parameters to replace or supplement intensity and duration could be used to specify operation of the disintegrator 1. These could be received externally to the disintegrator, or specified by the microcontroller 34.

Feedback systems on many of the tasks performed throughout the DNA extraction process could be implemented. For example, a reed switch could be installed on the freezer's 41 pneumatic cylinders to confirm if the door is open/closed. Microswitches could be installed on the loading bay of the liquid handler 42 to ensure the correct positioning of the plates. Feedback control could also be installed on the tissue disintegrator to ensure correct placement of the samples, detect opening/closing of the clamp, and detect speed and position of the holder/rotating shaft.

Figure 9 shows another sample processing system for DNA extraction utilising an alternative preferred disintegrator which preferably operates with the same sequence of operations as set out in the flow diagram of Figure 8. Features and functionality can be considered to be the same as those described above unless described otherwise below, and like reference numerals are used to indicate like parts with the addition of 100. The general features of the integrator 101 are similar to those mentioned with reference to Figures 1 to 5, however the drive mechanism differs. In particular, the base plate 112 is cantilevered off a main body portion 112a, which main body portion 112a is slidably mounted on a pair of vertical slide bars 150. A drive shaft 138 extends from a motor 129 to an offset cam 128, which extends through an enlarged aperture in the main body portion 112a. When the motor 129 is operated, rotation of the offset cam within the enlarged aperture in the main body portion 112a causes the main body portion 112a to slide up and down on the vertical slide bars 150, thereby shaking a the tissue receptacle 20 clamped on the base plate 112.

The tissue disintegrator 101 is integrated with the liquid processing system for DNA (or other component) extraction (which may, for example, be a THEONYX system manufactured by MWG Biotech). The liquid processing system preferably includes a pair of robotic arms 145a, 145b each of which is operable to move a tool to any position on a work surface 151, which work surface is shown in phantom lines in the Figure.

The tools may be one or more liquid transfer tools such as pipetting tools 152 or one or more gripping tools 154. The gripping tool (s) is/are suitable for transferring plates, consumables and other resources to any position on the work surface 151, or to a peripheral device such as the tissue disintegrator 101 or cool storage device unit. In the embodiment shown, robotic arm 145b is operable to move the pipetting tool 152, and robotic arm 145a is operable to move the gripping tool 154.

During a typical operation, the gripping tool 154 is moved by the robotic arm 145a to collect a well plate containing tissue samples from the work surface 151. The well plate is transferred to the holder 110 of the tissue disintegrator 101, as indicated for well plate 120, where the well plate is clamped and the samples are shaken for the required duration. Following a typical time period of about 1 to 3 minutes in the tissue disintegrator, the samples within the well plate have been ground to a fine powder. The well plate is then transferred to the work surface 151 by the gripping tool 154.

Following the transfer, the pipetting tool 152 performs the remainder of the DNA extraction process which would typically take about 1 hour, by manipulating a number of aqueous solutions.

In one typical process, the samples will be suspended in a DNA extraction buffer, and then removed from the buffer. The samples are then mixed with magnetic beads, and transferred to a magnetic separator through the use of the gripping tool 154. Following magnetic separation, the pipetting tool 152 is then used to remove the remaining liquid, and a wash buffer is added to the beads. The wash buffer is then removed from the beads. This washing process could be repeated three times, for 11-ie beads are then suspended in water, and magnetically separated again. The end product liquid (which in this case is purified DNA) is removed and put in another well plate, and then transferred to the cool storage 141. In the embodiment shown, the cool storage unit 141 includes a number of hinged covers 141 a. It is possible for the tissue sample (s) to initially be positioned within the cool storage unit 141 and for one of the robotic arms to remove the sample (s) from the cool storage unit and deliver it/them to the holder.

The gripping tool 154 is utilised to transfer consumables to and from various positions/resources on the liquid handler's work surface 50 (e. g. to or from a magnetic separation device). A plate disposal region 156 is provided for disposal of the used well plates as required. It will be appreciated that the above described process is one option only for obtaining components such as DNA, and the system could be used for other processes. For example, the system could include a modified silica columns and a centrifuge to separate the DNA and purify the compounds components of interest.

The system can also include other devices for analysis techniques. For example, in the embodiment shown, the apparatus includes a PCR device 160 for PCR analysis of the samples.

It is preferred that at least the liquid processing system and robotic device are controlled by one controller. For example, in one embodiment a PC may be used to control the liquid handler, and the liquid handler software program could be configured to initiate an executable file which sends a command to the disintegrator controller to operate the clamp, motor, timing etc. When the disintegrator has completed its cycle the executable file finishes, allowing the liquid handler to proceed with its operations (including operation of the robotic device). It is possible that the liquid processing system, robotic device and disintegrator are all controlled by one controller.

The operation can be fully automated and can occur when the system is unattended, allowing increased productivity.

The above describes preferred embodiments only, and modifications may be made thereto without departing from the scope of the following claims.

For example, the holder is shown as having a clamp with a base part and a clamp member. Instead, the clamp could be configured to clamp the sides or ends of the receptacle. Alternatively, rather than using a clamp the holder could use other means to hold the receptacle-such as magnetic means for example.

The sample processing systems are described as being used to extract DNA from tissue samples. However, the system has applications for extracting other component (s) from other sample (s). For example, the component could be a molecule such as DNA or a protein, a compound, or an element for example. The samples could be organic or inorganic materials, such as biological tissue, paint chips, etc. The system has application for metabolomics, forensic, medicine, etc.