Patent ID: 12235197

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

In order to make the description of the present disclosure more detailed and complete, the following provides an illustrative description for the implementation of the present disclosure and specific embodiments; but this is not the only way to implement or use the specific embodiments of the present disclosure. The embodiments disclosed below can be combined or substituted with each other under beneficial circumstances, and other embodiments can also be added to an embodiment without further description.

In order to solve the issues of the prior art, the present disclosure provides an automatic processing device for liquid samples, thereby reducing labor costs, shortening the overall processing time, and avoiding human mistaking and human judgment errors. For example, the device can automatically process and identify multiple sets of liquid samples, so it can avoid human mistaking and subsequent execution errors. In addition, the device can automatically read, so it can avoid human judgment errors.FIG.1is a conceptual implementation top view of an automatic processing device for liquid samples according to some embodiments of the present disclosure. As shown inFIG.1, the device includes a sample region10, a control module20, an image identification device30and a centrifuge40.

In some embodiments, the device further includes a reagent region50for storing reagents (e.g., reagent A, reagent B, reagent C, reagent D, and reagent E) that need to be used in a method of processing liquid samples (e.g., a method of separating cells from blood). In some embodiments, the reagent region50may be an ambient temperature reagent region or a refrigerated reagent region.

In some embodiments, the device further includes a culture device region60for placing culture devices602, such as culture dishes or multi-well culture dishes.

In some embodiments, the device further includes an operating panel70, which is coupled to the control module20. The operator can issue instructions through the operating panel70so that the control module20can operate according to the instructions.

In some embodiments, the device further includes a sterilization device (not shown), such as an ultraviolet lamp, which can irradiate ultraviolet rays in the operating space to achieve a sterilization effect.

The sample region10is configured to accommodate a plurality of tubes, such as liquid sample tubes (e.g., blood collection tubes)100and/or tubes101,102,103,104. The sample in the liquid sample tube100may be from a human or animal body, such as a cat, dog, or another mammal that can be raised. The tubes101,102,103, and104are, for example, 15 ml centrifuge tubes or 50 ml centrifuge tubes, which may be empty tubes or tubes filled with reagents. In some embodiments, the tube101contains the reagent A and the reagent B, and the tube103contains the reagent A, and the tubes102and104are empty tubes. In some embodiments, the device further includes a warning system (not shown). If the number of samples is odd, the device cannot be activated.

The control module20includes a mechanical module22. In some embodiments, the mechanical module22includes a mechanical arm and electric pipettes, which are configured to move the blood collection tubes100and/or the tubes101,102,103,104, and to unscrew and tighten (screw) upper caps of the blood collection tubes100and/or the tubes101,102,103,104, and to draw or discharge samples or reagents. In some embodiments, the device further includes a carrier (not shown), which is coupled to the control module20and configured to carry and move the blood collection tubes100and/or the tubes101,102,103, or104in the sample region10to facilitate the operations of the mechanical module22.

The image identification device30is coupled to the control module20. The image identification device32in the image identification device30is configured to identify a height of a blood volume in each of the blood collection tubes100. The next step can be performed if the heights of the blood volumes in the blood collection tubes100are the same. In some embodiments, if the heights of the blood volumes in the blood collection tubes100are inconsistent, a set value is used as a reference (e.g., the operator can set the height of the blood volume through the operating panel70). The reagent B may be taken out through the mechanical module22and added to the blood collection tube100that the height of the blood volume does not reach the set value, so that the height of the blood volume in each of the blood collection tubes100reaches the set value, so that subsequent centrifugal separation treatment can be performed. In some embodiments, the image identification device32includes an automatic optical inspection (AOI) system. In some embodiments, the automatic optical inspection system uses machine vision to record positions of the samples, sampling determination, and operation processes.

The centrifuge40is coupled to the control module20. The centrifuge40is configured to perform the centrifugal separation treatment on the samples in the centrifuge tubes (e.g., the tubes101,102,103, or104). In some embodiments, the centrifuge40includes a positionable system that can be integrated and tested to make the position of the sample consistent before and after the centrifugal treatment, so as to prevent the sample from being erroneously taken after the centrifugal separation treatment. In some embodiments, the centrifuge40has an oscillating function, which can help the sample and the reagent to be uniformly mixed.

In some embodiments, the image identification device34in the image identification device30is configured to capture images of the samples in the tubes after centrifugal treatment.

In some embodiments, the image identification device36in the image identification device30is configured to identify a position of a certain layer of the sample in the tube. In some embodiments, the image identification device36is combined with the mechanical module22to completely take out the layer of the sample.

The control module20is coupled to and configured to control the operation of the mechanical module22, the image identification device30, and the centrifuge40to perform various steps of the method of processing the liquid samples.

FIG.2is an actual implementation top view of an automatic processing device for liquid samples according to some embodiments of the present disclosure.FIG.3is an actual implementation front perspective view of an automatic processing device for liquid samples according to some embodiments of the present disclosure.FIG.4is an actual implementation rear perspective view of an automatic processing device for liquid samples according to some embodiments of the present disclosure.FIG.5is an actual implementation schematic diagram of an appearance of an automatic processing device for liquid samples according to some embodiments of the present disclosure. As shown inFIGS.2to4, the device includes a sample region10, a mechanical module22, an image identification device30, and a centrifuge40.

As shown inFIGS.2and3, the sample region10is configured to accommodate the liquid sample tubes100and the centrifuge tubes101,102,103, and104. In some embodiments, the sample region10includes a placement region10a, an uncapping/capping region10badjacent to the placement region10a, and a liquid drawing/discharging region10cadjacent to the uncapping/capping region10b.

The mechanical module22is configured to unscrew or tighten upper caps of the centrifuge tubes101,102,103, and104, and is configured to draw liquid from the centrifuge tubes101,102,103or104or discharge liquid to the centrifuge tubes101,102,103or104. In some embodiments, the centrifuge tubes at least includes a first group of the centrifuge tubes (e.g., the tubes101or102) and a second group of the centrifuge tubes (e.g., the tubes103or104), and a size of each of the first group of the centrifuge tubes is larger than that of each of the second group of the centrifuge tubes. In some embodiments, the centrifuge tubes at least includes two sets of the first group of the centrifuge tubes (e.g., the tubes101and102), and the two sets of the first group of the centrifuge tubes have the same size.

As shown inFIGS.2to4, the mechanical module22includes an uncapping/capping machine222and a liquid drawing/discharging machine224. The uncapping/capping machine222is configured to simultaneously unscrew or tighten upper caps of the first group of the centrifuge tubes (e.g., the tubes101or102), and is configured to simultaneously unscrew or tighten upper caps of the second group of the centrifuge tubes (e.g., the tubes103or104). The liquid drawing/discharging machine224is configured to simultaneously draw the liquid from the first group of the centrifuge tubes (e.g., the tubes101or102), and is configured to simultaneously draw the liquid from the second group of the centrifuge tubes (e.g., the tubes103or104), and is configured to simultaneously discharge the liquid to the first group of the centrifuge tubes, and is configured to simultaneously discharge the liquid to the second group of the centrifuge tubes. For example, the liquid drawing/discharging machine224can simultaneously draw the liquid from the first group of the centrifuge tubes (e.g., the tubes102) and then simultaneously discharge the liquid to the second group of the centrifuge tubes (e.g., the tubes103). The liquid drawing/discharging machine224can also simultaneously draw the liquid from the first group of the centrifuge tubes (e.g., the tubes101) and then simultaneously discharge the liquid to another first group of the centrifuge tubes (e.g., the tubes102).

In some embodiments, as shown inFIGS.2and3, the uncapping/capping machine222includes a plurality of clamping portions2222. As such, the clamp portions2222of the uncapping/capping machine222can clamp upper caps of at least one group of tubes (e.g., the liquid sample tubes100and/or the centrifuge tubes101,102,103or104) of the placement region10a. The group of the tubes can be moved to the uncapping/capping region10bby the uncapping/capping machine222, and then uncapped.

In some embodiments, as shown inFIGS.2and3, the device further includes a carrier105, which is configured to carry at least one group of the tubes (e.g., the liquid sample tubes100and/or the centrifuge tubes101,102,103or104), and can move back and forth between the uncapping/capping region10band the liquid drawing/discharging region10c. In some embodiments, after the group of the tubes is uncapped, the carrier105moves from the uncapping/capping region10bto the liquid drawing/discharging region10cto move the group of the tubes to the liquid drawing/discharging region10cfor performing subsequent liquid drawing and discharging steps. After the liquid drawing and discharging steps are performed, the carrier105moves from the liquid drawing/discharging region10cto the uncapping/capping region10bto move the group of the tubes to the uncapping/capping region10bfor performing subsequent capping steps.

The image identification device30is coupled to the control module. The image identification device30can identify a height of a blood volume in each of the blood collection tubes100, and take images of the samples in the tubes before and/or after the centrifugal treatment, and/or identify a position of a certain layer of the sample in each of the tubes. In some embodiments, the device further includes an image identification device38, which can identify and confirm data of a label of the tube (e.g., the liquid sample tube100) to avoid subsequent execution errors.

The centrifuge40is coupled to the control module. In some embodiments, the device further includes a horizontal oscillator42. In some embodiments, the horizontal oscillator42is disposed adjacent to the centrifuge40.

In some embodiments, as shown inFIGS.2and3, the device further includes a reagent region50for storing reagents (e.g., the reagents501,502,503,504and505).

In some embodiments, as shown inFIGS.2and4, the device further includes a culture device region60for placing culture devices602, such as culture dishes or multi-well culture dishes. In some embodiments, the device further includes a culture temporary storage region62for temporarily storing the culture devices602.

In some embodiments, as shown inFIGS.2and4, the device further includes a consumable region80configured to accommodate consumables such as micropipettes.

In some embodiments, as shown inFIG.5, the device further includes an operating panel70, which is coupled to the control module. The operator can issue instructions through the operating panel70to make the mechanical module22, the image identification device30and the centrifuge40coupled to the control module operate according to the instructions.

In some embodiments, as shown inFIGS.2and5, the device further includes an outer cover90, which covers the sample region10, the mechanical module22, the image identification device30, the centrifuge40, the reagent region50, the culture device region60and the consumable region80to prevent the external environment from affecting the test operation. In some embodiments, the outer cover90includes a window90a, and the inside of the device can be cleaned through the window90a. The operator can also put the samples, the reagents, and the consumables to be tested into the device through the window90a. In some embodiments, the device further includes an air filter92, such as a high-efficiency particulate air (HEPA), to ensure the cleanliness of the operating environment.

It is worth noting that the uncapping/capping machine222shown inFIG.3can uncap upper caps of each of the multiple groups of tubes with different sizes, and various embodiments thereof will be described in detail below.FIG.6is a schematic diagram of an uncapping/capping machine according to some embodiments of the present disclosure. As shown inFIG.6, the clamping portion2222of the uncapping/capping machine222has a first clamping portion2222aand a second clamping portion2222b, and the second clamping portion2222bis located over the first clamping portion2222a. The first clamping portion2222acorresponds to an upper cap of one of the first group of the centrifuge tubes (e.g., the tubes101or102), and the second clamping portion2222bcorresponds to an upper cap of one of the second group of the centrifuge tubes (e.g., the tubes103or104). In some embodiments, a width W2of a second space defined by the second clamping portion2222bis smaller than a width W1of a first space defined by the first clamping portion2222a. Since the clamping portion2222of the uncapping/capping machine222has the above-mentioned structure, it can be configured to simultaneously unscrew (or tighten) the upper caps of the first group of the centrifuge tubes and can simultaneously unscrew (or tighten) the upper caps of the second group of the centrifuge tubes.

In some embodiments, as shown inFIGS.3and6, the uncapping/capping machine222is further configured to pull up upper caps of a group of the liquid sample tubes (e.g., the blood collection tubes100). In some embodiments, the clamping portion2222of the uncapping/capping machine222further has a third clamping portion2222clocated over the second clamping portion2222b, and the third clamping portion2222ccorresponds to an upper cap of one of the liquid sample tubes (e.g., the blood collection tubes100). In some embodiments, a width W3of a third space defined by the third clamping portion2222cis smaller than the width W2of the second space defined by the second clamping portion2222b. Since the clamping portion2222of the uncapping/capping machine222has the above-mentioned structure, it can be configured to simultaneously unscrew (or tighten) the upper caps of the first group of the centrifuge tubes and can be configured to simultaneously unscrew (or tighten) the upper caps of the second group of the centrifuge tubes, and it can also be configured to simultaneously pull up (or press back) the upper caps of the group of the liquid sample tubes.

In another aspect, the liquid drawing/discharging machine224shown inFIG.3can freely adjust the spacing between the micropipettes to match the spacing of each of the multiple groups of tubes with different sizes. Various embodiments thereof will be described in detail below.FIG.7Ais a schematic diagram of a liquid drawing/discharging machine according to some embodiments of the present disclosure.FIG.7Bis a schematic diagram of a roller of a liquid drawing/discharging machine according to some embodiments of the present disclosure. As shown inFIG.7A, the liquid drawing/discharging machine224includes a roller2242and a plurality of micropipettes2244, and the micropipettes2244are coupled to the roller2242. In some embodiments, the liquid drawing/discharging machine224can automatically unload the used micropipettes2244, and then install new micropipettes2244.

As shown inFIG.7B, the roller2242includes a plurality of grooves2242gspirally surrounding the roller2242. The grooves2242gare configured to be respectively coupled to the micropipettes2244shown inFIG.7A. Each of the grooves2242ghas a first end2242aand a second end2242b. A spacing between the first ends2242aof the grooves2242gis smaller than a spacing between the second ends2242bof the grooves2242g. As such, the roller2242can be rotated to adjust the spacing between the micropipettes2244coupled to the grooves2242g.

In some embodiments, the grooves2242ginclude a first group of the grooves (e.g., the four grooves on the left as shown inFIG.7B) and a second group of the grooves (e.g., the four grooves on the right as shown inFIG.7B), and the first ends2242aof the first group of the grooves are away from the first ends2242aof the second group of the grooves. In some embodiments, the roller2242has two ends2242topposite to each other, and the first ends2242aof the first group of the grooves and the first ends2242aof the second group of the grooves are close to the two ends2242t, respectively. As such, before the liquid drawing and discharging steps, the micropipettes2244shown inFIG.7Acan be located beneath the two ends2242tof the roller2242, and the space beneath a middle section of the roller2242can be freed to avoid the micropipettes2244taking up too much space.

The following provides a method of automatically separating cells from blood using the above-mentioned device. In some embodiments, the above-mentioned device is suitable for automatically separating peripheral blood mononuclear cells from a blood sample of a biological body.FIG.8Ais a schematic diagram of pre-test processing of a method of automatically separating cells from blood according to some embodiments of the present disclosure.FIGS.9to15are schematic diagrams of a method of automatically separating cells form blood according to some embodiments of the present disclosure.

First, the reagents501,502,503,504,505, the blood collection tubes100, the tubes101,102,103,104and the consumables required for the method of separating cells from blood are put into the device. Subsequently, the pre-test treatment of the tubes101is performed. As shown inFIGS.2and8A, the tubes101(each of the tubes101has a filter membrane (not labeled)) are moved to the uncapping/capping region10bby the uncapping/capping machine222of the mechanical module22, and the upper caps are then unscrewed, and the tubes101are then moved to the draw/discharge region10cby the carrier105. The reagent A (501) is added to the tubes101, and the tubes101are then moved to the uncapping/capping region10bby the carrier105, and the upper caps are then tightened, and the tubes101are then moved to the centrifuge40by the uncapping/capping machine222for the centrifugal separation treatment. After the centrifugal separation treatment is completed, the tubes101are taken out by the uncapping/capping machine222of the mechanical module22and placed back into the uncapping/capping region10b. Next, the upper caps of the tubes101are unscrewed by the uncapping/capping machine222, and the tubes101are then moved to the liquid drawing/discharging region10cby the carrier105, and the reagent B (502) is then added to the tubes101. In some embodiments, the step of adding the reagent A (501) or that of adding the reagent B (502) mentioned above can be performed using a micro pump (not shown), a liquid drawing pipette (not shown) and a liquid discharging pipette (not shown) connected to the micro pump to draw out the reagent A (501) or the reagent B (502) and added it into the tubes.

Next, a test is carried out. As shown inFIGS.2and9, the upper caps of the blood collection tubes100are pulled up by the uncapping/capping machine222, and the blood collection tubes100are then moved to the liquid drawing/discharging region10cby the carrier105, and the height of the blood volume in each of the blood collection tubes100is identified by the image identification device30. If the heights of the blood volumes in the blood collection tubes100are the same, the next step is performed; if the heights of the blood volumes in the blood collection tubes100are inconsistent, a set value is used as a reference, and the reagent B (502) may be discharged into the blood collection tube100, so that the height of the blood volume in each of the blood collection tubes100reaches the set value.

As shown inFIGS.2and10, the liquid in the blood collection tubes100is respectively added to the tubes101through the liquid drawing/discharging machine224.

As shown inFIG.11, the reagent B (502) is added to the blood collection tubes100for cleaning, and the liquid in the blood collection tubes100is then added to the tubes101, respectively. In some embodiments, this step is optional and can be repeated.

As shown inFIGS.2,11and12, the upper caps of the tubes101are tightened, and the tubes101are then moved to the centrifuge40for the centrifugal separation treatment, which may be also called as a first centrifugal separation treatment.

As shown inFIGS.2and12, after the centrifugal separation treatment is completed, the tubes101are taken out from the centrifuge40and placed back into the uncapping/capping region10b. The upper caps of the tubes101and the tubes102are unscrewed, and the supernatant401in the tubes101is drawn and then discharged into the tube102. In some embodiments, after the centrifugal separation treatment is completed, images of the tubes101after the centrifugal separation treatment are taken by the image identification device30. In some embodiments, the image identification device30can not only capture the images after the centrifugal separation treatment of the tubes101is completed, but also determine whether the sample is completely separated after the centrifugal separation treatment, and whether hemolysis has occurred.

As shown inFIG.13, the reagent B (502) is added to the tubes101for cleaning, and the liquid in the tubes101is then added to the tubes102, respectively. In some embodiments, this step is optional and can be repeated.

As shown inFIGS.2,13and14, the upper caps of the tubes102are tightened, and the tubes102are then moved to the centrifuge40for the centrifugal separation treatment, which may also be called as a second centrifugal separation treatment.

As shown inFIGS.2and15, after the centrifugal separation treatment is completed, the tubes102are taken out from the centrifuge40and placed back into the uncapping/capping region10b. The upper caps of the tubes102are unscrewed, and a portion of the supernatant402in the tubes102are removed, leaving some of the supernatant402(e.g., about 1 ml), and the upper caps of the tubes102are then tightened. The samples in the tubes102are the peripheral blood mononuclear cells.

In some embodiments, the above-mentioned device is also suitable for automatically separating circulating tumor cells (CTCs) from a blood sample of a biological body. In some embodiments, the circulating tumor cells separated by the method can be cultured and expanded, and the expanded circulating tumor cells can be tested for drug screening to provide medication guidance for subsequent clinical diagnosis.

The method of separating the circulating tumor cells from blood will be described in detail below.FIG.8Bis a schematic diagram of pre-test processing of a method of automatically separating cells from blood according to some embodiments of the present disclosure.FIGS.16to19are schematic diagrams of a method of automatically separating cells form blood followingFIG.15according to some embodiments of the present disclosure.

First, the pre-test treatments of the tubes101and the tubes103are performed. For the pre-test treatment of the tubes101, please refer to the description ofFIG.8Aabove, so it will not be repeated here. For the pre-test treatment of the tubes103, as shown inFIG.8B, the upper caps of tube103are unscrewed, and the reagent A (501) is added to tubes103, and the upper caps of the tubes103are then tightened.

Next, as shown inFIGS.2,15and16, the reagent C (503) is drawn and discharged into the tubes102, and the upper caps of the tubes102are them tightened, and the tubes102are then moved to the horizontal oscillator42for the oscillation treatment, so that the sample and the reagent C in each of the tubes102are uniformly mixed. In some embodiments, after the reagent C is added to the tubes102, the tubes102stand for a period of time, and the oscillation treatment is then performed.

As shown inFIGS.2and17, after the oscillation treatment is completed, the reagent D (504) is taken and added to the tubes102. In some embodiments, the liquid drawing/discharging machine224ofFIG.7Ais used to draw and discharge the sample and the reagent D in the tubes102to make them uniformly mixed in the tubes102.

As shown inFIGS.17and18, the liquid in the tubes102is slowly added to the tubes103(containing the reagent A (501)) along the walls of the tubes103respectively, so that the sample in the tube102falls on top of the reagent A due to the density difference and thus those present complete layers, so as to prevent the sample in the tube102from falling into the reagent A and mixing with it. In some embodiments, the discharging speed and the discharging position (e.g., close to the tube wall) of the micropipette2244of the liquid drawing/discharging machine224are controlled to achieve the above purpose.

As shown inFIGS.2,18, and19, the tubes103are moved to the centrifuge40for the centrifugal separation treatment, which may also be called as a third centrifugal separation treatment. In some embodiments, after the centrifugal separation treatment is completed, the image identification device30captures images after the centrifugal separation treatment of the tubes103is completed.

As shown inFIG.19, after the centrifugal separation treatment is completed, the image identification device30is configured to identify intermediate layers, which may also be called as cell layers, to be taken out of the tubes103, respectively, and the intermediate layers are then taken out from the tubes103by the liquid drawing/discharging machine224and then added to the tubes104. In some embodiments, the image identification device30identifies the intermediate layer to be taken out through the color difference.

Next, the image identification device30is configured to identify a height of a liquid volume in each of the tubes104. If the heights of the liquid volumes in the tubes104are the same, the next step can be performed; if the heights of the liquid volumes in the tubes104are inconsistent, a set value is used as a reference, and the reagent D (504) is discharged into the tube104, so that the height of the blood volume in each of the tubes104reaches the set value. Next, the upper caps of the tubes104are taken up by the mechanical module22and tightened, and the tubes104are then moved to the centrifuge40for the centrifugal separation treatment, which may also be called as the fourth centrifugal separation treatment.

After the centrifugal separation treatment is completed, the supernatant in the tubes104is removed, and the remaining samples in the tubes104are the purified circulating tumor cells.

In some embodiments, a sufficient amount of the reagent E (e.g., more than or equal to 10 ml of the reagent E, which may be a culture medium) is added to the tubes104. In this way, the circulating tumor cells inside the tubes104can be stored for a longer period of time.

In some embodiments, an appropriate amount of the reagent E (e.g., 3 ml to 5 ml of the reagent E, which may be a culture medium) is added to the tubes104by the mechanical module22. Subsequently, the sample (including the purified circulating tumor cells and the culture medium) in the tube104can be added to the culture device602in the culture device region60by the liquid drawing/discharging machine224to perform expansion of the circulating tumor cells.

In some embodiments, the operator can select one of the above-mentioned two treatments (i.e., adding the sufficient amount of the reagent E to the tubes104and adding the appropriate amount of the reagent E to the tubes104) through the operating panel70of the device according to the purpose.

In some embodiments, the culture medium for culturing the circulating tumor cells at least includes basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). In some embodiments, the culture medium at least includes platelet lysate. In some embodiments, the culture medium at least includes MEM (or RPMI1640) and other suitable culture medium and antibiotics for avoiding contamination by microorganisms and fungi. The culture device602and the method of expanding the circulating tumor cells by the culture device602can refer to Taiwan Patent No. 1672376, U.S. Provisional Application No. 62/827,248 and U.S. Provisional Application No. 62/931,236.

It can be seen from the above that the use of the above device for automatically processing the liquid samples (e.g., automatically separating cells from blood) can reduce labor costs, shorten the overall processing time, and avoid human mistaking and human judgment errors. In addition, parameters of the above-mentioned device can be set and adjusted to conform to the steps of separating cells from blood, and thus can be widely used in any field that requires separating cells from blood.

The above-mentioned embodiments are merely illustrative to illustrate the principles and effects of the present disclosure, as well as to explain the technical features of the present disclosure, and are not configured to limit the scope of protection of the present disclosure. Any person skilled in the art can easily complete changes or equal arrangements without departing from the technical principle and spirit of the present disclosure, and all fall within the claimed scope of the present disclosure.

The above-mentioned embodiments are only illustrative of the principles and effects of the present disclosure, as well as explaining the technical features of the present disclosure, rather than limiting the scope of protection of the present disclosure. Anyone who is familiar with the technology can easily complete changes or equal arrangements without violating the technical principles and spirit of the present disclosure. All of them belong to the claim scope of the present disclosure.