CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of US Provisional Application No. 62/256,581, filed November 17, 2015, and US Provisional Application No. 62/334,614, filed May 1 1 , 2016, both of which are incorporated herein by reference.

INCORPORATION BY REFERENCE

[0002] All publications and patent applications mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

FIELD

[0003] Various embodiments of the disclosure relate generally to apparatus and methods for processing and preparing samples. Specifically, the disclosure relates to apparatus and methods for processing fluid and/or tissue samples with reagents, preparing a smear or a monolayer film.

BACKGROUND

[0004] Sample analysis is valuable in medical diagnosis, clinical research, agricultural development and environmental control. For another example, lung biopsy tissue sample analysis is important in diagnosis of pulmonary pathological processes, such as granulomas, neutrophilic infiltration, necrosis and miliary tuberculosis (TB), which are all related to respiratory mortality in children in the developing world.

[0005] For example, blood sample analysis is important in medical diagnosis and clinical research of many diseases. Malaria diagnosis via microscopy began in the late 1 800s and today remains the detection standard. Microscopy can provide a quantitative assessment of peripheral blood parasitemia and parasite stages. The blood samples need to be chemically processed and smeared before being viewed under a microscope. There are two kinds of smears. Thick smears are often used for diagnosis and quantitative assessment, while thin smears are for species identification. Typically, air-dried thin blood smear slides are fixed in methanol. Thick blood smears are not fixed; they comprise a thick layer of dehemoglobinized (lysed) red blood cells.

[0006] Conventionally, to prepare a thick blood film, a blood spot deposited on a slide is stirred in a circular motion using the corner of another slide and allowed to dry without fixative. Once dry, the blood is stained with diluted Giemsa for approximately 20 minutes and washed in buffered water for a few minutes. The slide is then air dried in a vertical position and examined using microscopy. To prepare a thin blood smear conventionally, the smooth edge of a slide is placed in a drop of blood on a second slide at a 45° angle and smeared quickly and steadily along the surface. After air-drying, the film is fixed with methanol for about 2 minutes and air-dried again. The sample is then stained with diluted Giemsa for 15-30 minutes and washed by dipping the slide in buffered water. Lastly, the slide is air-dried in a vertical position and examined with microscopy.

[0007] Today, field technicians prepare blood smears as best they can, often under sub-par laboratory standards. Samples are exposed to the ambient air for extended periods of time, promoting contamination, and the environment may not be ideal for preserving the smears until they are viewed under a microscope. There have been some proposals of automatic blood smearing apparatus. However, these apparatuses are usually bulky, complicated and expensive, thus not practical for field applications.

[0008] In addition, the conventional methods of blood processing and smearing are time- consuming and labor-intensive, and the quality of the blood sample smearing can be affected by technician-to-technician technique differences.

[0009] For another example, lung biopsy tissue sample analysis is important in diagnosis of pulmonary pathological processes, such as granulomas, neutrophilic infiltration, necrosis and miliary tuberculosis (TB), which are all related to respiratory mortality in children in the developing world. Conventionally, long standing tissue preparation practice need fixation, paraffin embedding, microtome-prepared samples, solvents including xylene and ethanol, as well as all the associated long processing time.

[0010] Similar problems exist for other samples processing and smearing. There is a need for a practical apparatus that can perform sample processing and smearing in the field. There is also a need for a novel method for sample processing and smearing that can save time and increase test efficiency as well as sample consistency. There is further a need to reduce preparation time, solvent use and sample contamination, while facilitating cost-effective pathological evaluation and analysis by minimally trained users in the field.

SUMMARY OF THE DISCLOSURE

[0011] The present disclosure relates to apparatus and methods for processing and smearing of samples. Specifically, the disclosure relates to an apparatus that is configured to chemically process a sample by mixing the sample with a reagent, smear a mixture of the sample with the reagent to create a smeared layer and be ready to be viewed and analyzed. Various embodiments of methods of processing and smearing of samples are also disclosed.

[0012] Various embodiments of a sample processing and smearing apparatus are disclosed. In general, the apparatus can comprise a sample receiver configured to receive a sample and a reagent reservoir configured to hold a reagent. The apparatus can further comprise a mixer which can be adapted and disposed to receive and mix the sample from the sample receiver and the reagent from the reagent reservoir into a mixture. The apparatus can further comprise a viewing chamber in fluid communication with the mixer to receive the mixture. The viewing chamber can comprise a viewing stage and a transparent cover over the viewing stage. The transparent cover can be movable with respect to the viewing stage between a sample inflow position and a smeared position. The mixture can be smeared on the viewing stage for viewing through the transparent cover by moving the transparent cover from the sample inflow position to the smeared position. The smeared mixture can form a monolayer film which can be viewed on the viewing stage directly without being transported to a conventional microscope slide.

[0013] In some embodiments, the sample is a fluid sample. In some embodiments, the sample receiver comprises a capillary tube. The sample can be collected by capillary action. The capillary tube can comprise a first opening extending through a distal end of the sample receiver for receiving the sample and a second opening in fluid communication with the mixer. The capillary tube can be extending from a neck of the main stage body in some embodiments. By using a capillary tube, the sample receiver can receive a controlled amount of the sample, for example, less than 50 μΐ.

[0014] In some embodiments, the apparatus further comprises a cap of the capillary tube. The cap can be adapted to cover the first opening of the capillary tube. The reagent reservoir can be disposed in the cap in some embodiments. In some embodiments, the apparatus further comprises a seal disposed between an interior surface of the cap and an exterior surface of the sample receiver.

[0015] In some embodiments, the reagent reservoir comprises a separator arranged and configured so that movement of the cap onto the sample receiver opens the separator to dispense the reagent from the reagent reservoir into the capillary tube. The separator can be configured to separate the reagent reservoir from the sample receiver. The separator can also be used to seal the reagent before mixing the reagent with the sample. In some embodiments, the separator can comprise a foil cover enclosing the reagent in the reagent reservoir in one end of the cap. In some embodiments, movement of the cap onto the sample receiver opens the separator to dispense the reagent from the reagent reservoir into the capillary tube. In some embodiments, the apparatus can further comprise a seal between an interior surface of the cap and an exterior surface of the sample receiver or a neck where the sample receiver is disposed in to seal the distal end.

[0016] In some embodiments, the cap and the sample receiver are dimensioned such that the distal end of the sample receiver extends through the separator when the cap is placed on the sample receiver. Movement of the cap onto the sample receiver can move the reagent and the sample through the mixer and into the viewing chamber. In some embodiments, the volume of the reagent reservoir plus the volume of the sample receiver is configured to be greater than the primary sample volume inside the main stage body. The mixer can have a funnel shape to assist a turbulent flow and promote mixing.

[0017] In some embodiment, the apparatus further comprises a microfiuidic channel between the static mixer and the viewing chamber and a microfiuidic channel between the sample receiver and the static mixer. Microfiuidic lines are employed in distribution of the fluid sample or the mixture of the fluid sample and the reagent in some embodiments. The apparatus can also comprise an overflow chamber in some embodiments.

[0018] The sample processing and smearing apparatus can a fully enclosed apparatus, thus eliminating the possibility of the sample being contaminated by environment and bio-hazards from infectious diseases. The apparatus can perform the sample collection, chemical processing of the sample by mixing the sample with the reagent, and smear preparation of the mixture of the sample and the reagent. In addition, the smeared monolayer of the mixture can be viewed on the viewing stage of the apparatus directly without being transported to the conventional microscope slides.

[0019] In some embodiments, the sample comprises a tissue sample. In some embodiments, the sample receiver is configured to receive a sample acquisition needle. In some embodiments, the apparatus comprises a main stage body, wherein the reagent reservoir is disposed inside the main stage body. In some embodiments, the reagent reservoir is located beneath the viewing stage. In some embodiments, the apparatus further comprises a one-way valve.

[0020] In some embodiments, the sample receiver and reagent reservoir are disposed and arranged such that injection of the sample into the sample receiver leads to mixing the reagent with the sample.. In some embodiments, wherein injection of the sample into the sample receiver leads to entry of a mixture of the sample and the reagent onto the viewing stage. In some embodiments, the reagent reservoir comprises a separator arranged and configured such that a movement of the sample acquisition needle into the sample receiver opens the separator to dispense the reagent from the reagent reservoir. In some embodiments, a movement of the sample acquisition needle onto the sample receiver moves the reagent and the sample through the mixer and onto the viewing stage. In some embodiments, a movement of the transparent cover from the sample inflow position to the smeared position reveals a thin stained tissue section.

[0021] Various embodiments of methods of processing and smearing a sample are also disclosed. In general, the methods can comprise receiving a sample in a sample receiver, mixing the sample with a reagent in a mixer in fluid communication with the sample receiver, and moving a mixture of the sample and the reagent into a viewing chamber in fluid communication with the mixer, wherein the viewing chamber comprises a transparent cover. The method can further comprise moving the transparent cover from a sample inflow position to a smeared position to spread the mixture and viewing the mixture on a viewing stage of the viewing chamber.

[0022] In some embodiments receiving the sample comprises moving the sample through the sample receiver by capillary action. In some embodiments receiving the sample comprises receiving a controlled amount of the sample. In some embodiments mixing the sample with a reagent comprises placing a cap on the sample receiver to pierce a separator of a reagent reservoir disposed in the cap. In some embodiments moving a mixture of the sample and the reagent comprises placing a cap on the sample receiver to pierce a separator of a reagent reservoir and moving the reagent and the sample into the viewing chamber. In some

embodiments moving the transparent cover from a sample inflow position to a smeared position comprising lowering the transparent cover.

[0023] In general, the methods can comprise moving the transparent cover from a sample inflow position to a smeared position to spread the mixture. The methods can further comprise viewing the smeared mixture on a viewing stage of the viewing chamber directly. In some embodiments, the methods can comprise receiving the sample through a sample receiver by capillary action. The methods can further comprise receiving a controlled amount of the sample.

[0024] In some embodiments, the methods can comprise mixing the sample with a reagent comprising placing a cap on a sample receiver to pierce a separator of a reagent reservoir. In some embodiments, the methods can further comprise pushing the reagent into the sample. In some embodiments, the methods can comprise mixing the sample with the reagent in a mixer by a turbulent flow. In some embodiments, the mixer can be in a funnel shape to assist the mixing by the turbulent flow. In some embodiments, the methods can comprise distributing a mixture of the sample and the reagent by pushing a cap on a sample receiver to pierce a separator of a reagent reservoir. The methods can further comprise distributing the mixture through a microfluidic path. In some embodiments, the methods can comprise pushing the mixture to the viewing chamber by configuring the volume of the reagent reservoir plus the volume of the sample receiver to be greater than the primary sample volume inside the main stage body. [0025] In some embodiments, the methods can comprise lowering the transparent cover to smear the mixture to create a monolayer film. The methods can further comprise viewing the smeared mixture on a viewing stage of a fully enclosed chamber. In general, the apparatus can be viewed directly under a microscope. The apparatus can also be used with fluorescence imaging. The apparatus can further be viewed for colorimetry reaction, exothermic reaction, or endothermic reaction for a variety of analytical and diagnostic purposes.

[0026] In general, the sample processing and smearing apparatus can be a fully enclosed device. The apparatus can be configured to perform sample collection, reagent mixing, sample distribution and smearing. The apparatus can be used for processing and smearing a variety of samples including blood, saliva, urine, cerebrospinal fluid, liquid biopsy, contaminated water, etc. A variety of reagents can be used to mix with the samples to create chemical reactions in the apparatus.

[0027] In general, various embodiments in this disclosure comprise a fully enclosed, disposable apparatus that collects an exact amount of sample, chemically processes and stains the sample, smears the mixture and enables viewing and analyzing. Various embodiments disclose apparatus and methods of receiving the sample by using capillary action and distributing the sample by microfluidic channels. The disclosure also provides apparatus and methods of smearing the mixture of the sample and the reagent and viewing the smeared mixture in the same viewing chamber.

[0028] Disclosed herein is a sample processing and smearing apparatus. The apparatus can comprise a sample receiver configured to receive a sample acquisition needle. The sample acquisition needle can contain a tissue sample. The apparatus can comprise a main stage body, a reagent reservoir disposed inside the main stage body to store a reagent, and a viewing chamber in fluid communication with the sample receiver disposed and configured to receive the sample and the reagent. The viewing chamber can comprise a viewing stage and a transparent cover over the viewing stage, the transparent cover being movable with respect to the viewing stage between a sample inflow position and a smeared position. A mixture of the sample and the reagent is smeared on the viewing stage for viewing through the transparent cover by moving the transparent cover from the sample inflow position to the smeared position. In some

embodiments, the apparatus can further comprise a one-way valve.

[0029] In some embodiments the reagent reservoir is located beneath the viewing stage. In some embodiments a movement of injecting of the sample into the sample receiver leads to mixing the reagent with the sample. In some embodiments a movement of injecting of the sample into the sample receiver leads to entry of a mixture of the sample and the reagent onto the viewing stage. In some embodiments the reagent reservoir comprises a separator, and wherein a movement of the sample acquisition needle into the sample receiver opens the separator to dispense the reagent from the reagent reservoir. In some embodiments a movement of the sample acquisition needle onto the sample receiver moves the reagent and the sample through the mixer and onto the viewing stage. In some embodiments a movement of the transparent cover from the sample inflow position to the smeared position reveals a thin stained tissue section.

[0030] Disclosed herein is a method of processing and smearing a tissue sample. The method can comprise receiving a sample acquisition needle containing a tissue sample into the sample receiver. The method can comprise mixing the sample with a reagent in a mixer in fluid communication with the sample receiver. The method can comprise moving a mixture of the sample and the reagent into a viewing chamber in fluid communication with the mixer, wherein the viewing chamber comprises a transparent cover. The method can comprise moving the transparent cover from a sample inflow position to a smeared position to spread the mixture. The method can comprise viewing the mixture on a viewing stage of the viewing chamber. In some embodiments the reagent reservoir is located beneath the viewing stage.

[0031] In some embodiments the receiving step comprises injecting the sample from the sample acquisition needle into the sample receiver to cause mixing the reagent with the sample. In some embodiments the receiving step comprises further comprising injecting the sample from the sample acquisition needle into the sample receiver to cause movement of a mixture of the sample and the reagent onto the viewing stage.

[0032] In some embodiments the method further comprises opening a separator of the reagent reservoir to dispense the reagent from the reagent reservoir. In some embodiments the method further comprises moving the sample acquisition needle onto the sample receiver to move the reagent and the sample through the mixer and onto the viewing stage. In some embodiments the method further comprises moving the transparent cover from the sample inflow position to the smeared position to reveal a thin stained tissue section. In some embodiments the method further comprises using a one-way valve to control an amount of a portion of the sample to be injected into the apparatus.

[0033] In some embodiments the method further comprises injecting the sample to lead to mixing the reagent with the sample. In some embodiments the method further comprises injecting the sample to lead to entry of a mixture of the sample and the reagent onto the viewing stage. In some embodiments the method further comprises opening a separator of the reagent reservoir to dispense the reagent from the reagent reservoir. In some embodiments the method further comprises g moving the sample acquisition needle onto the sample receiver to move the reagent and the sample through the mixer and onto the viewing stage. In some embodiments the method further comprises moving the transparent cover from the sample inflow position to the smeared position to reveal a thin stained tissue section. In some embodiments the method further comprises using a one-way valve to portion the tissue sample.

BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The novel features of the invention are set forth with particularity in the claims that follow. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative

embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0035] FIG. 1 is a perspective view of a sample processing and smearing apparatus according to one embodiment of the disclosure.

[0036] FIG. 2 is a section view of the sample processing and smearing apparatus when a transparent cover is in a sample inflow or raised position.

[0037] FIG. 3 is a section view of the sample processing and smearing apparatus when the transparent cover is in a smeared or lowered position.

[0038] FIG. 4A is a top view of a prototype of the sample processing and smearing apparatus after being assembled.

[0039] FIG. 4B is a top view of the foil inside a cap of the prototype of the apparatus.

[0040] FIG. 5A illustrates the prototype of the sample processing and smearing apparatus in close proximity to a finger prick blood drop.

[0041] FIG. 5B illustrates the capillary tube of the apparatus filled with blood after touching the blood drop.

[0042] FIG. 5C illustrates a cap being pushed on the capillary tube, where a separator in the cap is pierced to release the dye.

[0043] FIG. 5D illustrates the dye and the blood sample is distributed to a viewing chamber after being mixed in the mixer by a turbulent flow.

[0044] FIG. 5E illustrates the transparent cover is moving from the sample inflow or raised position to the smeared or lowered position and a smear layer is forming.

[0045] FIG. 5F illustrates a smear layer after the transparent cover is actuated by the user to the smeared or lowered position.

[0046] FIG. 5G is a block diagram of the method of processing and smearing the sample.

[0047] FIG. 6A is an example of 20x photo taken through the transparent cover of the sample processing and smearing apparatus of red blood cells in an approximate monolayer.

[0048] FIG. 6B is an example of 50x photo taken through the transparent cover of the sample processing and smearing apparatus of red blood cells in an approximate monolayer. [0049] FIG. 7 A is an optical image of the patterned PDMS stamp with 10 μπι raised squares.

[0050] FIG. 7B illustrates a second PDMS stamp.

[0051] FIG. 8A schematically illustrates a perspective view of a sample processing and smearing apparatus 800 according to another embodiment of the disclosure.

[0052] FIG. 8B schematically illustrates a section view of the sample processing and smearing apparatus 800 in FIG. 8A.

[0053] FIG. 9A illustrates lung tissue with granuloma stained mast cells.

[0054] FIG. 9B illustrates lung tissue with toluidine blue stained mast cells. DETAILED DESCRIPTION

[0055] The present disclosure relates to apparatus and methods for processing and smearing of samples. Specifically, the disclosure relates to an apparatus that is configured to process a sample, smear and create a monolayer of the sample film and be ready to be viewed and analyzed.

[0056] The apparatus can comprise a sample receiver configured to receive a sample, a reagent reservoir, a mixer where the sample can be mixed with the reagent, and a viewing chamber. The mixer can be adapted and disposed to receive and mix the sample from the sample receiver and the reagent from the reagent reservoir into a mixture. The viewing chamber in fluid

communication with the mixer to receive the mixture. The viewing chamber can comprise a viewing stage and a transparent cover over the viewing stage. The transparent cover can be movable with respect to the viewing stage to between a sample inflow position and a smeared position. The mixture can be smeared on the viewing stage to form a monolayer film for viewing through the transparent cover by moving the transparent cover from the sample inflow position to the smeared position. The apparatus can be a fully enclosed device, thus eliminating possible sample contamination and environmental hazards. The apparatus can be used for a sample matrix analysis. For example, the sample can include blood, tissue, saliva, urine, cerebrospinal fluid, liquid biopsy, contaminated water, etc.

[0057] The apparatus can further be viewed and analyzed directly under a microscope without transporting the sample to conventional microscope slides. The apparatus can also be used with fluorescence imaging. The apparatus can further be viewed for colorimetry reaction, exothermic reaction, or endothermic reaction for a variety of analytical and diagnostic purposes.

[0058] The apparatus can be configured to perform sample collection, reagent mixing, sample distribution and smearing. By incorporating sample chemical processing and smearing into one practical apparatus, the efficiency of the sample diagnosis and analysis can be increased. The apparatus does not require power and an individual with minimal training is able to safely and accurately operate the apparatus. Therefore, sample consistency can be increased. Testing sensitivity and specificity may also be increased.

[0059] The apparatus can be used to observe live cells in a sample. For example, the apparatus can facilitate blood smear acquisition and viewing for live cell malaria testing. In addition, a variety of blood, solution or tissue tests including blood cells counts, fluorescent testing, cancer detection, FISH assays, water testing, soil testing, minimally invasive autopsy processing, etc. may be afforded or partially afforded with the apparatus. Furthermore, it is not necessarily a requirement that cells be alive. In general, the apparatus can be used to observe thin smear or thick smear sample films whether the cells are alive or not.

[0060] In general, the apparatus can be used to process and prepare a variety of samples.

Though blood sample and tissue sample are used as examples, it is important to note, however, that the apparatus is not limited to process and prepare the blood sample and the tissue sample. The apparatus can be used to process and smear saliva, urine, cerebrospinal fluid, liquid biopsy, contaminated water and other samples as well.

[0061] Similarly, a variety of reagents can be mixed with the samples to create chemical reactions. The dye/stain is used as an example of reagents in the following description, but the apparatus is not limited to use the dye/stain as a reagent. Various dyes/stains can be used to mix with the blood sample and stain the blood sample. Giemsa and Leishman are two of the most common dyes that can be used to mix with the blood sample. Giemsa plus 8% Dimethyl sulfoxide (DMSO) can used to allow differential staining of white blood cells (WBC) vs. red blood cells (RBC) for supravital staining (live cells). Methylene blue typically does not require cell fixing before staining and therefore can be used for supravital stain. Toluidine blue is another strong enough stain to penetrate RBC and stain malarial parasites. Other dyes/stains can be used to be mixed with the blood sample as well.

[0062] The apparatus can be fully enclosed and disposable. The apparatus can be configured to collect an exact amount of sample, chemically process and stain the sample, smear the mixture and enable viewing and analyzing. In some embodiments, the apparatus can be configured to use capillary draw and microfluidic channels. Capillary draw and channel fabrication can provide sufficient capillary force to draw a significant amount of the sample suitable to form a monolayer film. In some other embodiments, the apparatus can be configured to receive a sample acquisition needle. For example, the apparatus can be configured to receive a single use core biopsy device such as BARD MONOPTY disposable Core biopsy instrument. The apparatus can further comprise a viewing chamber that is configured to prepare the smearing layer as well as view the smeared film. [0063] FIG. 1 schematically illustrates a perspective view of a sample processing and smearing apparatus 100 according to one embodiment of the disclosure. FIG. 2 schematically illustrates a section view of the sample processing and smearing apparatus 100. The apparatus 100 can comprise a sample receiver 2, a reagent reservoir 4, a main stage body 3, a mixer 6 and a transparent cover 15 covering a viewing stage 16. The main stage body 3 can have a substantial cuboid shape as shown in FIG. 1. However, the man stage body 3 can have other shapes such as cylindrical, cubic, etc. The main stage body 3 can have a height between about 1 mm to about

30 mm, a width between about 5 mm to about 50 mm and a length between about 5 to about 100 mm. For example, the main stage body 3 can have a height between about 5 mm to about 15 mm, a width between about 25 mm to about 45 mm and a length between about 30 to about 50 mm. Values outside the above ranges are also possible. The dimensions of the main stage body 3 can be changed to meet the need. The dimensions of the main stage body 3 can depend on the need of the dimension of the viewing stage which will be discussed below.

[0064] In some embodiments, the apparatus 100 can further comprise an overflow chamber 20 to collect the waste. The overflow chamber 20 can be enclosed in the main stage body 3. The overflow chamber 20 can have a height between about 1 mm to about 20 mm, a width between about 5 mm to about 40 mm and a length between about 5 to about 30 mm. Values outside the above ranges are also possible. For example, the overflow chamber 20 can have a height between about 1 mm to about 8 mm, a width between about 5 mm to about 15 mm and a length between about 5 to about 20 mm. Values outside the above ranges are also possible. The dimensions of the overflow chamber 20 can be depended on the dimensions of the main stage body 3 and can be changed to meet the need. In some other embodiments, the overflow chamber 20 may not be necessary.

[0065] Referring to FIG. 1 and FIG. 2, the sample receiver 2 can comprise a capillary tube 2 in some embodiments. The capillary tube 2 can comprise a first opening 2b extending through a distal end 2c of the sample receiver 2 for receiving the sample and a second opening 2d in fluid communication with the mixer 6. The sample receiver 2, for example, the capillary tube, can be configured to receive the sample by capillary action. The sample collection via the capillary action can be configured to intake a controlled amount of the blood sample. For example, the capillary tube 2 can be a micro-capillary tube designed to intake a dictated volume of blood. The dictated volume or controlled amount of blood can be about 0.5, 1 , 1.5, 2, 2.5, 3, 5, 10, 15, 20, 30, 40, 50 μΐ or any values therebetween. Values outside the above range are also possible. The dictated volume or controlled amount of blood can be changed to meet the need. The capillary tube 2 can have a length between about 1 mm to about 50 mm. The capillary tube 2 can have a length between about 10 mm to about 30 mm in some embodiments. Values outside the above range are also possible. The dimensions of the capillary tube 2 can be changed to meet the need.

In some other embodiments, a negative pressure can be incorporated to facilitate the blood sample collection. In yet some other embodiments, some other flow control can be incorporated to collect the sample. In still some other embodiments, the sample receiver 2 can simply be an inlet to receive the sample.

[0066] As shown in FIG. 1 and FIG. 2, the apparatus can further comprise a neck 8. The neck 8 can be protruding from a side of the man stage body 3. The sample receiver 2 can be partially disposed inside the neck 8, where the front portion 2a of the sample receiver 2 is exposed and extending from the neck 8. The neck 8 can have a substantial cylindrical shape as shown in FIG. 1. The neck 8 can have other shapes as well. The length of the neck 8 can be between about 0 mm to about 25 mm. The length of the neck 8 can be between about 2 mm to about 20 mm in some embodiments. Values outside the above ranges are also possible. The dimensions of the neck 8 can be changed to meet the need. In some other embodiments, the neck 8 can be eliminated.

[0067] In some embodiments, the sample receiver 2 can comprise a capillary tube 2 as discussed above. The front portion 2a of the capillary tube 2 can be completely exposed from the neck 8 as shown in FIG. 2. A seal 9 can be disposed on an exterior surface of the neck 8. The seal 9 can be used to seal the first opening 2b extending through the distal end 2c of the capillary tube 2. In some other embodiments, the capillary tube 2 can be partially disposed inside the main stage body 3 with the front portion 2a extending from the main stage body 3 without the neck 8. The seal 9 can be disposed on an exterior surface of the capillary tube 2. The seal 9 can be used to seal the first opening 2b at the distal end 2c of the capillary tube 2.

[0068] The apparatus 100 can further comprise a cap 5 adapted to cover the first opening 2b of the capillary tube, wherein the reagent reservoir 4 can be disposed in the cap 5. The cap 5 can be configured to be put over the neck 8 and the front portion 2a of the capillary tube 2 so that an interior surface of the cap 5 engages a seal 9 to seal the open distal end 2c of the capillary tube 2. The reagent reservoir 4 can be disposed inside a tip or a front portion 5a of the cap 5.

[0069] The apparatus 100 can comprise the reagent reservoir 4 configured to be filled with a reagent, for example, a dye to be used to stain the blood sample. In one embodiment, the reagent reservoir is disposed in the cap 5 of the capillary tube 2. The cap 5 can comprise three cylindrical sections, the front section or the tip 5a, a middle section 5b and a back section 5c. The front section or the tip 5a has the smallest diameter among the three sections and the reagent reservoir 4 is disposed in the front section or the tip 5a of the cap 5. The diameter of the reagent reservoir 4 is configured to be slightly bigger than the outer diameter of the capillary tube 2 such that the capillary tube 2 can be inserted into the reservoir 4 when the cap 5 is pushed on the capillary tube 2. The middle portion 5b of the cap is configured to match the outer diameter of the neck 8 and to engage the seal 9 in the neck 8 after the cap 5 is pushed on the apparatus 100. The back portion 5c is configured to be put on a back portion of the neck 8 to fix the cap 5 on the apparatus 100. The back portion 5c can be screwed on, or clicked on the back portion of the neck 8, or connected to the neck 8 by other mechanical connecting methods. The cap 5 can have a length between about 1 mm to about 30 mm. For example, the length of the cap 5 can be between about 10 mm to 25 mm. Values outside the above range are also possible. The length of the cap 5 can be changed to meet the need. In some other embodiments, the cap 5 can be in other shapes or have more or less portions, not limited to the example configurations or dimensions described above.

[0070] The cap 5 can further comprise a separator 7 which is configured to separate the dye and the blood sample. The separator 7 is configured to hold the dye in the tip 5a of the cap 5 as shown in FIG.2. For example, the separator 7 can comprise a foil cover in some embodiments. The dye can be stored in the reservoir 4 behind the foil cover7. The reservoir 4 and the separator 7 can be disposed in the cap 5 in some embodiments.

[0071] After the blood already being collected in the capillary tube 2, a user can place the cap 5 onto the capillary tube 2 when holding the neck 8 of the apparatus 100. The cap 5 and the sample receiver 2 can be dimensioned such that the distal end 2c of the sample receiver 2 extends through the separator 7 when the cap 5 is placed on the sample receiver 2. The movement of pushing the cap 5 on by the user can pierce the separator 7 and force the dye in the reservoir 4 to flush through the capillary tube 2, and both the blood and the dye can be pushed through a first micro fluidic channel 12 to the mixer 6, then through a second microfluidic channel 13 towards the viewing chamber 10.

[0072] The first microfluidic channel 12 can be disposed at a base 6a of the apparatus and between the capillary tube 2 and the base 6a. The first microfluidic channel 12 can have a circular cross-section or a substantial circular cross-section in some embodiments. The first microfluidic channel 12 can have a rectangular or a substantial rectangular or square cross- section in some other embodiments. In some alternate embodiments, the cross-section of the first microfluidic channel 12 can be in any other shapes, not limited to circular or rectangular or square shapes. The shortest dimension of the cross section of the first microfluidic channel 12 can be between about 50 nm to about 1000 μιη. For example, the shortest dimension of the cross section of the first microfluidic channel 12 can be between about 200 microns to about 800 microns. For example, the first microfluidic channel 12 can have a circular cross-section with a diameter about 500 microns in some embodiments. Values outside the above range are also possible. The dimensions of the first microfluidic channel 12 can be changed to meet the need. [0073] In some embodiments, the first microfluidic channel 12 can comprise two portions. As shown in FIG. 2, the first portion 12a can be disposed vertically and in fluid communication with the second opening 2d of the capillary tube 2. The second portion 12b can be disposed horizontally and in fluid communication with the mixer 6. The length of the first portion 12a can be between about 0 mm to about 10 mm, and the length of the second portion 12b can be about 0 mm to about 30 mm in some embodiments. For example, the length of the first portion 12a can be between about 1 mm to about 8 mm, and the length of the second portion 12b can be about 1 mm to about 20 mm in some embodiments. Values outside the above range are also possible.

The length of the. first microfluidic channel 12 can be changed to meet the need. In some other embodiments, the first portion 12a is not necessary. In yet some other embodiments, the second portion 12b is not necessary. In some alternative embodiments, the first microfluidic channel 12 can be disposed in any directions. In some other embodiments, the first microfluidic channel 12 can be eliminated, and the sample receiver 2 can be connected to the mixer 6 directly.

[0074] In some other embodiments, the reservoir 4 can be disposed inside the main body 3. The separator 7 of the reservoir 4 can be used to separate the sample and the reagent. After the sample is collected by the sample receiver 2, the separator 7 can be opened by pierced such that the sample and the reagent can the mixed.

[0075] The apparatus 100 can further comprise the mixer 6. The mixer 6 can be disposed close to the viewing chamber. In one embodiment, the mixer 6 can be disposed below the viewing chamber 10 on a base 6a of the apparatus 100. In another embodiment, the mixer 6 can be disposed adjacent to the viewing chamber 10. The mixer 6 and the viewing chamber 10 are configured to be in fluid communication. For example, the mixer 6 and the viewing chamber 10 can be connected by a second microfluidic channel 13. The second microfluidic channel 13 can have a circular cross-section or a substantial circular cross-section in some embodiments. The second microfluidic channel 13 can have a rectangular or a substantial rectangular or square cross-section in some other embodiments. In some alternate embodiments, the cross-section of the second microfluidic channel 13 can be in any other shapes, not limited to circular or rectangular or square shapes. The shortest dimension of the cross section of the second microfluidic channel 13 can be between about 50 nm to about 1000 μιη. In some embodiments, the shortest dimension of the cross section of the second microfluidic channel 13 can be between about 200 microns to about 800 microns. For example, the second microfluidic channel 13 can have a circular cross-section with a diameter about 500 microns in some embodiments. Values outside the above range are also possible. The dimensions of the cross-section of the second microfluidic channel 13 can be changed to meet the need. The length of the second microfluidic channel 13 can be between about 0 mm to about 30 mm in some embodiments. For example, the length of the second microfluidic channel 13 can be between about 4 mm to about 12 mm in some embodiments. Values outside the above range are also possible. The length of the second microfluidic channel 13 can be changed to meet the need. In some other embodiments, the second microfluidic channel 12 can be eliminated, and the mixer 6 can be connected to the viewing chamber 10 directly.

[0076] The mixer 6 is configured to collect the blood first and does not allow the blood to reach the viewing chamber 10 until the blood has mixed with the dye that arrives in the mixer 6 just after the blood does. The volume of the dye reservoir 4 plus the volume of the blood sample can be configured to be greater than the primary sample volume inside the main stage body 3, which can ensure that the blood sample and dye mixture reaches the viewing chamber 10. The primary sample volume inside the main stage body 3 can include the volume of the capillary tube, the volume of the first microfluidic channel 12 between the capillary tube 2 and the mixer 6, the volume of the mixer 6 and the volume of the second microfluidic channel 13 between the mixer 6 and the viewing chamber 10.

[0077] The mixer can be in an inverted funnel shape as shown in FIG. 2. The funnel shape can assist in the formation of turbulent flow. In some embodiments, the mixer 6 can have a height between about 0.1 mm to about 10 mm, and a diameter on a base 6a between about 0.1 mm to about 10 mm. For example, the mixer 6 can have a height between about 1 mm to about 3 mm, and a diameter on the base 6a between about 1 mm to about 2 mm. Values outside the above range are also possible. The dimensions of the mixer 6 can be changed to meet the need. The shape and design of the mixer 6 can have many variations. For example, the mixer 6 can also be in a pyramid shape in some other embodiments. The mixer 6 can have any other shapes such as cylindrical or cubic in yet some other embodiments. After the mixer 6 fills with the mixture, it allows the mixture to proceed up to the viewing chamber 10.

[0078] By pushing the dye filled cap 5 on, the blood and dye can be driven into the mixer 6 and up onto the viewing chamber 10 by turbulent flow force. The blood and the dye are pushed to the mixer by the first microfluidic channel 12. The mixer 6 can be specifically configured to facilitate the mixing of the blood sample and the dye. A turbulent flow can be generated when the blood sample and the dye have chemical reaction in the mixer 6. After the targeted volume of blood plus dye arrives in the mixer 6, the dye and the blood mix via turbulent flow and then the mixture of the blood and the dye is released upwards and onto the viewing chamber 10. Pushing the cap on is only step required by the user to move samples and mix blood.

[0079] Referring to FIG. 2 and FIG.3, the viewing stage 16 can be disposed at a center of a top side of the main stage body 3. The top surface of the view stage 16 can be in a circular shape in some embodiments. The top surface of the viewing stage 16 can be in rectangular, square, elliptical, or any other shapes in some other embodiments. The apparatus 100 can further comprise a viewing chamber 10 between the transparent cover 15 and the viewing stage 16 as shown in FIG. 2. The diameter or the shortest dimension of a top surface of the viewing stage 16 can be between about 1 mm to about 30 mm. For example, the diameter or the shortest dimension of a top surface of the viewing stage 16 can be between about 10 mm to about 20 mm in some embodiments. Values outside the above range are also possible. The dimensions of the viewing stage 16 can be related to the dimension of the main stage body 3 and can be changed to meet the need.

[0080] The transparent cover 15 can be in circular shape in some embodiments. In some other embodiments, the transparent cover 15 can be in rectangular, square or other various shapes depending on the shape of the viewing stage 16. The diameter or the shortest dimension of the transparent cover 15 can be between about 2 mm to about 30 mm. For example, the diameter or the shortest dimension of the transparent cover 15 can be between about 10 mm to about 20 mm in some embodiments. Values outside the above range are also possible. The dimensions of the transparent cover 15 can depend on the dimensions of the viewing stage 16 and can be changed to meet the need.

[0081] The transparent cover 15 can comprise an O-ring 17 at an outer edge to create a seal of the viewing chamber 10. The transparent cover 15 can have a sample inflow or "open" position and a smeared or "closed" position. When the transparent cover 15 is in the sample inflow or "open" position, the cover 15 is raised above the viewing stage 16 as shown in FIG 2. When the transparent cover 15 in the sample inflow or "open" position, there is a slight space between the transparent cover 15 and the viewing stage 16. The sample inflow position allows the inflow of the mixture of the blood sample and the dye. The smeared or "closed" position allows the monolayer of the mixture films to be viewed. The apparatus 100 is a fully sealed device when the transparent cover 1 is in the smeared or "closed" position.

[0082] FIG. 3 schematically illustrates the apparatus 100 when the transparent cover 15 is in the smeared or "closed" position. The transparent cover 15 is movable between the sample inflow or raised or "open" position and the smeared or lowered or "closed" position. During the process when the transparent cover 15 is moving to the smeared or "closed" position, the mixture of the blood and dye is smeared. The excessive mixture can be pushed to the overflow chamber 20. The overflow chamber 20 can reduce pressure and catch any excess blood that may need to leave the viewing stage 16 upon actuation of the transparent cover 15 in order to create the smear layer. When the transparent cover 15 is in the smeared or "closed" position, the smeared mixture of the blood sample and the dye can be viewed through a microscope or by a naked eye. [0083] The range of the movement of the transparent cover 15 can be configured to create the desired smear layer. In one embodiment, the transparent cover 15 can be lowered down on top of the blood plus dye solution, creating a 'smear layer' comprised of multilayers of cells near the periphery of the circular stage 16 and approximate monolayers on blood cells on the bulk of the stage 16. In another embodiment, the transparent cover 15 can be pushed down to a first "closed" position to create a thick smear, and be pushed down to a second "closed" position to create a thin smear. The movement of the transparent cover 15 (closing) can be accomplished by pressing down with fingers, squeezing, rotating or snapping closed to a dictated position that gives of a "click" sound when done, or by rotating the cover 15 to the "closed" position.

[0084] FIG. 4A illustrates an assembled prototype of the sample processing and smearing apparatus. The dye can be seen in the cap 5 and the dye is held back by the foil 7. The capillary tube 2 protrudes from the neck 8 of the stage body 3. The first microfiuidic channel 12 and second microfiuidic channel 13 can be visualized on the base 6a of apparatus. The mixer 6 is right under the viewing stage 16.with the foil inside the cap holding back dye/ stain. FIG. 4B illustrates the foil cover 7 inside a cap 5 which is configured to hold back the dye.

[0085] The sample process and smearing apparatus 100 can be made from injection moldable materials, such as polypropylene, but the apparatus 100 can also be made from acrylic, silicone materials, etc. In some embodiments, the apparatus 100 can be made from opaque materials. In some embodiments, the transparent cover 15 of the apparatus 100 can be made from materials that are opaque to some light wavelengths, but transparent to some other wavelengths.

[0086] Various embodiments of the disclosure also disclose novel methods of processing and smearing a sample, for example, a blood sample. Conventional methods of manually performing thin smear or thick smear can be time consuming and the smear can be easily contaminated by the environment. FIGS. 5A to 5G illustrate the novel methods of processing and smearing a blood sample.

[0087] The method can comprise receiving a controlled amount of a sample, for example, a blood sample. FIG. 5A illustrates the prototype of the sample processing and smearing apparatus in close proximity to a finger prick blood drop. The user can collect the blood sample by capillary action in some embodiments. For example, the user can use the sample processing and smearing apparatus and place the apparatus in close proximity to a finger prick blood drop as shown in FIG. 5A.

[0088] FIG. 5B illustrates the capillary tube of the apparatus filled with blood after touching the blood drop. The controlled amount of blood sample can be collected by capillary action as shown in FIG. 5B. [0089] The method can comprise mixing the sample with a reagent, for example, mixing a blood sample with a dye. The user can mix the blood sample with the dye by pushing a cap filled with the dye to the capillary tube of the apparatus. FIG. 5C illustrates a cap being pushed on the capillary tube, where a separator in the cap is pierced to release the dye. The blood sample and the dye can be mixed in a mixer in the apparatus.

[0090] The method can further comprise distributing the mixture of the sample and the reagent to a viewing chamber, for example, distributing the mixture of the blood sample and the dye to a viewing chamber. FIG. 5D illustrates the dye and the blood sample is distributed to a viewing chamber after being mixed in the mixer by turbulent flow. The chemical reaction of the blood sample and the dye can generate turbulent flow, resulting in the mixture being released onto the viewing stage of the viewing chamber. The transparent cover of the apparatus having a sample inflow or "open" position to allow the mixture to flow in and a smeared or "closed" position to view the smeared blood sample. The transparent cover is in sample inflow or "open" position as shown in FIG. 5D.

[0091] The method can comprise smearing the mixture on the viewing stage of the viewing chamber. The method can comprise lowering the transparent cover to spread the mixture and create the smear layer in some embodiments. FIG. 5E illustrates the transparent cover is moving from the "open" position to the "closed" position and a smear layer is forming. FIG. 5F illustrates a smear layer after the transparent cover is actuated by the user to the "closed" position.

[0092] FIG. 5G is a block diagram of the method 500 of processing and smearing the sample. The method 500 can comprise receiving a sample 510. In some embodiments, the method can comprise receive the sample through a sample receiver by capillary action 510a in some embodiments.

[0093] The method can comprise mixing the sample with a reagent 520. In some embodiments, the methods can comprise mixing the sample with a reagent comprising pushing a cap on a sample receiver to pierce a separator of a reagent reservoir 520a in some embodiments.

[0094] The method can further comprise distributing a mixture of the sample and the reagent to a viewing chamber 530. The methods can further comprise distributing the mixture through a microfluidic channel 530a in some embodiments.

[0095] The method can further comprise moving a transparent cover of the viewing chamber 540 from a sample inflow position to a smeared position to spread the mixture. The method can further comprise lowering a transparent cover to smear the mixture in some embodiments. The methods can further comprise viewing the smeared mixture on a viewing stage of the viewing chamber 550. [0096] FIG. 6A is an example of 20x photo of red blood cells in an approximate monolayer taken through the transparent cover 15 of the apparatus 100. FIG. 6B is an example of 50x photo of red blood cells in an approximate monolayer taken through the transparent cover 15 of the apparatus 100. The average diameter of the red blood cells is about 8 microns and the average diameter of the white blood cells varies, and is always bigger than the average diameter of the red blood cells.

[0097] The sample process and smearing apparatus 100 can have many variations and forms without departure from the spirit and scope of the disclosure. For example, the apparatus can further comprise a lancet so that a separate finger prick device is not required for blood collection. The lancet can be disposed at a tip of the sample receiver in some embodiments. In some other embodiments, the apparatus can further comprise markings to denote insertion directionality or other information of the sample or the reagent.

[0098] In some embodiments, the apparatus 100 can comprise an inlet to receive a sample and a reagent. In some embodiments, the apparatus 100 can comprise a first inlet for the sample, and a second inlet for the reagent. In yet some other embodiments, the apparatus 100 can comprise preloaded reagent inside. In yet some other embodiments, the sample and the reagent can be received through removing the transparent cover or by a hole in the transparent cover. In some embodiments, the apparatus can comprise a microfluidic channel and a viewing chamber in fluid communication with the microfluidic channel to receive the sample and the reagent. The microfluidic channel can be disposed to receive and distribute the fluid sample and the reagent to the viewing chamber directly. A mixture of the sample and the reagent can be smeared on the viewing stage for viewing through the transparent cover by moving the transparent cover from the sample inflow position to the smeared position.

[0099] In some embodiments, the sample and the reagent can be pushed directly to the viewing chamber and mixed in the viewing chamber by a turbulent flow from the chemical reaction, and then the mixture can be smeared in the same viewing chamber, while the excessive mixture can be released to the overflow chamber.

[0100] In yet some other embodiments, the transparent cover 15 can be sliding on from one side of the apparatus 100 instead of lowering down to spread the mixture and create the thin smear films. The transparent cover 15 can be configured to be movable from a sample inflow position to a smeared position by sliding along.

[0101] In still some other embodiments, the apparatus can further comprise another inlet to receive methanol, which can be used to fix the cells to the viewing stage. For example, another step and chamber may be added to the apparatus. Alternatively, methanol could have been incorporated in such a way as to expose the blood to methanol or a methanol vapor pressure in a chamber before staining.

[0102] In some other embodiments, the viewing stage 16 of the apparatus 100 can be patterned with Polydimethylsiloxane (PDMS) materials. The patterned PDMS can be fabricated by first making a patterned silicon wafer master via microlithography and associated reactive ion etching (RIE). The etched wafer can be cleaned up in a sulfuric acid/ hydrogen peroxide Piranha etch bath and then treated with a fluorosilane to render it hydrophobic. For example, uncured and degassed PDMS can be poured on top of the silicon master and baked at 60C for 1.5 hr. After cooling, the PDMS can be peeled off the wafer and diced up into smaller pieces with a razor blade. The PDMS can have the inverse topographical pattern of the silicon master.

[0103] Patterning the stage can assist forming a monolayer. By placing the patterned PDMS stamp on top of the mixture of the blood sample and the dye, the red blood cells (RBCs) may be forced into a regular matrix one cell thickness deep as shown in FIG. 7A. Visualization of stained parasites in such a configuration may make imaging processing easy. Forced cell lysis may be beneficial on the basis that RBCs are less permeable than white blood cells (WBCs). If cells are packed into a monolayer and lysed, the dye (for example, Giemsa) may be able to access parasites faster and more easily. FIG. 7B illustrates a second PDMS stamp used to pattern RBCs with 10 micron squares and slightly larger squares within squares. More individual monolayer cells are present; fewer cells are aggregated as shown in FIG. 7B.

[0104] FIG. 8A schematically illustrates a perspective view of a sample processing and smearing apparatus 800 according to another embodiment of the disclosure. FIG. 8B schematically illustrates a section view of the sample processing and smearing apparatus 800 in FIG. 8A. The sample processing and smearing apparatus 800 can be configured to receive or accept a sample acquisition needle. For example, the apparatus can be configured to receive a single use core biopsy device such as BARD MONOPTY disposable Core biopsy instrument. For example, the sample acquisition needle can contain lung biopsy specimens, which will be stained in the fully enclosed apparatus. The sample processing and smearing apparatus 800 can be configured to enable diagnosis of pulmonary pathological processes, such as granulomas, neutrophilic infiltration, necrosis and miliary tuberculosis, which are related to respiratory mortality in children in the developing world. The apparatus 800 can be a fully enclosed, disposable device that simplifies tissue processing from needle biopsies for use by non-experts in the field. The apparatus 800 can be configured to eliminate dependence on processing solvents and completely encloses the sample, protecting both the sample and the person preparing the sample while drastically decreasing sample preparation time, number of steps involved, and dependence on organic solvents. [0105] Referring to FIG. 8A and FIG. 8B, the apparatus 800 can comprise a sample receiver 802, a reagent reservoir 804, a main stage body 803, a mixer 806, a viewing chamber 810 and a transparent cover 815 covering a viewing stage 816. The main stage body 803 can have a substantial cuboid shape as shown in FIG. 8A. The apparatus 800 can comprise the viewing chamber 810 between the transparent cover 815 and the viewing stage 816 as shown in FIG. 8B . In some embodiments, the apparatus 800 can further comprise an overflow chamber 820 to collect the waste. The overflow chamber 820 can be enclosed in the main stage body 803. The sample receiver 802 can be configured to receive, or accept a sample acquisition needle 888. For example, the apparatus 800 can be configured to receive a single use core biopsy device such as BARD MONOPTY disposable Core biopsy instrument. The sample receiver 802 can comprise a capillary tube in some embodiments. The sample receiver 802 can comprise a first opening extending through a distal end of the sample receiver 802 for receiving the sample acquisition needle 888 and a second opening in fluid communication with the mixer 806.

[0106] As shown in FIG. 8A and FIG. 8B, the apparatus 800 can further comprise a neck 808. The neck 808 can be protruding from a side of the man stage body 803. The sample receiver 802 can be partially disposed inside the neck 808, where the front portion of the sample receiver 802 is exposed and extending from the neck 808. The neck 808 can have a substantial cylindrical shape as shown in FIG. 8A. The neck can have other shapes as well. In some embodiments, the neck 808 can be configured to receive, or accept the sample acquisition needle 888, for example, a single use core biopsy device such as BARD MONOPTY disposable Core biopsy instrument.

[0107] The reagent reservoir 804 can be located beneath the stage 803, such that injecting the sample into the apparatus 800 can lead to the sample mixing with dye and entry of dye-plus- tissue sample onto the viewing stage 816.

[0108] The apparatus 800 can comprise the reagent reservoir 804 configured to be filled with a reagent, for example, a dye to be used to stain the tissue sample. In one embodiment, the reagent reservoir 804 is disposed beneath the stage 803. In some embodiments, the reagent reservoir 804 can further comprise a separator 807 which is configured to separate the dye and the tissue sample. The separator 807 can be configured to hold the dye in the reagent reservoir 804 as shown in FIG. 8B. For example, the separator 807 can comprise a foil cover in some

embodiments. The dye can be stored in the reservoir 804 behind the foil cover 807. The reagent reservoir 804 can be disposed in the neck 808. The reagent reservoir 804 can have a variety of types, configuration, chambers and shapes, such as those described in US Patent Application No. 14/971 ,947, filed on December 16, 2015, which is incorporated by reference herein in its entirety. [0109] After the tissue sample has already been collected in the needle 888, a user can insert the needle 888 onto the sample receiver 802. The movement of injecting the sample into the apparatus 800 by pushing the needle 888 can pierce the separator 807 and force the dye in the reservoir 804 to move with the sample through a first microfluidic channel 812 to the mixer 806, then through a second microfluidic channel 813 towards the viewing chamber 810.

[0110] In some other embodiments, the reservoir 804 can be disposed inside the neck 808. In yet some other embodiments, the reservoir 804 can be disposed inside the main body 803. The separator 807 of the reservoir 804 can be used to separate the sample and the reagent. After the sample is collected by the sample receiver 802, the separator 807 can be opened by pierced such that the sample and the reagent can the mixed.

[0111] The apparatus 800 can further comprise the mixer 806. The mixer 806 and the viewing chamber 810 are configured to be in fluid communication. By pushing the needle 888, the tissue and dye can be driven into the mixer 806 and up onto the viewing chamber 810, e.g., by turbulent flow force. The tissue and the dye are pushed to the mixer 806 by the first microfluidic channel 812. The mixer 806 can be specifically configured to facilitate the mixing of the tissue sample and the dye. A turbulent flow can be generated when the tissue sample and the dye have a chemical reaction in the mixer 806. After the targeted volume of tissue plus dye arrives in the mixer 806, the dye and the tissue mix via turbulent flow, and then the mixture of the tissue and the dye is released upwards and onto the viewing chamber 810. In this embodiment, pushing the needle 888 is the only step required by the user to move samples and mix dye. Depressing the transparent cover 815 can reveal a thin stained tissue section.

[0112] In some embodiment, the apparatus 800 can further comprise a one-way valve (not shown) in the interior of the apparatus that only accepts a portion of the tissue sample. For example, sometimes, only a portion of the tissue is needed for processing and smearing, thus the needle 888 will need to be retracted from this device after this portion of the sample is injected into the apparatus. The one-way valve can be configured to only allow the sample to flow in one direction. The one-way valve can be configured to control the amount of the portion to be injected into the apparatus, and prevent this portion to flow out of the apparatus as the needle is retracting. In some embodiments, a cap can be used to enclose the apparatus after the needle is retracted from the apparatus.

[0113] The apparatus 800 can be configured to work with a variety of reagents, such as stains with surrogate lung material. The apparatus 800 can be further configured to work with desired tissue layer thickness. The apparatus 800 may lead to successful staining of known, targeted cells, such as mast cells, and the ability to view thin section tissue samples in the apparatus 800 between 20x and l OOx. The apparatus 800 can be configured to be usable by minimally trained, field-based healthcare workers and relies on only a few user-driven steps. The apparatus 800 can eliminate the need for fixation, paraffin embedding, microtome-prepared samples, solvents including xylene and ethanol, as well as the associated processing time. Samples can be quickly processed and imaged within the apparatus 800 at the point of need.

[0114] The apparatus 800 can create a thin, stained layer of lung tissue for evaluation of pulmonary pathological processes as shown in FIG. 9A and FIG. 9B. FIG. 9A illustrates lung tissue with granuloma stained mast cells. FIG. 9B illustrates lung tissue with toluidine blue stained mast cells. Stained, flash frozen needle biopsy samples have unique morphology as compared to fixed and paraffin embedded sample preparations, but that differences do not interfere with diagnosis.

[0115] In some embodiments, the apparatus 800 can be configured to collect the tissue sample and prepare for analysis within a minute, as opposed to a few hours by using conventional apparatuses. The apparatus 800 can produce resultant images within the apparatus that are at least equivalent to those acquired conventionally with painstaking preparation, solvents and imaging.

[0116] In other embodiments, the apparatus 800 can be compatible with any type of tissue under investigation, and it can be integrated it into point of need operations to reduce material and infrastructure requirements. Samples can be read onsite using a fieldable microscope (such as, e.g., the microscope described in US Patent Application No. 62/234,709, filed April 19, 2016, the disclosure of which is incorporated herein by reference) using embedded third-party machine learning algorithms. Sample results with image analysis, diagnosis, and other supporting data like subject location, environment, and demographic information can be sent to the cloud from the fieldable microscope.

[0117] Various embodiments of apparatuses and methods disclosed herein can result in dramatic simplification of long standing sample preparation practice and a drastic reduction in cost and time required for preparing such pathological samples from minimally invasive tissue collection for pathological evaluation.

[0118] While the present disclosure has been disclosed in example embodiments, those of ordinary skill in the art will recognize and appreciate that many additions, deletions and modifications to the disclosed embodiments and their variations may be implemented without departing from the scope of the disclosure.

[0119] A wide range of variations to those implementations and embodiments described herein are possible. Components and/or features may be added, removed, rearranged, or combinations thereof. Similarly, method steps may be added, removed, and/or reordered. [0120] Likewise various modifications to the implementations described in this disclosure may be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other implementations without departing from the spirit or scope of this disclosure. Thus, the claims are not intended to be limited to the implementations shown herein, but are to be accorded the widest scope consistent with this disclosure, the principles and the novel features disclosed herein.

[0121] Accordingly, reference herein to a singular item includes the possibility that there are a plurality of the same items present. More specifically, as used herein and in the appended claims, the singular forms "a," "an," "said," and "the" include plural referents unless specifically stated otherwise. In other words, use of the articles allow for "at least one" of the subject item in the description above as well as the claims below.

[0122] Additionally as used herein, a phrase referring to "at least one of a list of items refers to any combination of those items, including single members. As an example, "at least one of: a, b, or c" is intended to cover: a, b, c, a-b, a-c, b-c, and a-b-c.

[0123] Certain features that are described in this specification in the context of separate embodiments also can be implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single embodiment also can be

implemented in multiple embodiments separately or in any suitable subcombination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a subcombination or variation of a subcombination.

[0124] Similarly, while operations may be described as occurring in a particular order, this should not be understood as requiring that such operations be performed in the particular order described or in sequential order, or that all described operations be performed, to achieve desirable results. Further, other operations that are not disclosed can be incorporated in the processes that are described herein. For example, one or more additional operations can be performed before, after, simultaneously, or between any of the disclosed operations. In certain circumstances, multitasking and parallel processing may be advantageous. Moreover, the separation of various system components in the embodiments described above should not be understood as requiring such separation in all embodiments, and it should be understood that the described program components and systems can generally be integrated together in a single product or packaged into multiple products. Additionally, other embodiments are within the scope of the following claims. In some cases, the actions recited in the claims can be performed in a different order and still achieve desirable results.