RU2246349C2 - Testing board with many through passages for high-production screening - Google Patents

Abstract

FIELD: high-production screening of many specimens under test.

SUBSTANCE: proposed testing board has pair of opposite surfaces and many passages. Each passage runs from one opposite surface to other. Passages are combined in groups; each group has at least two "horizontal" rows and two "vertical" rows of passages. Groups are combined in sets; each set has at least two "horizontal" and two "vertical" rows of groups. For analyzing the solution, at least one opposite surfaces of testing board is immersed in solution under test. Portion of solution is admitted to each passage through hole in submerged surface. After passages are filled with solution, testing board is removed from solution and is kept above bearing surface. Solution is held in each passage due to surface tension. Solution contained in one or several passages is subjected to analysis and identification. Position of identified solution is marked according to its set or group.

EFFECT: simplified procedure of analysis.

22 cl, 9 dwg

Description

The technical field of the invention

The present invention generally relates to a test device and, in particular, to a test tablet with multiple pass-through channels for high throughput screening.

State of the art

Known test devices comprise a test plate having a pair of opposing surfaces and a plurality of wells. Wells extend from one of the opposite surfaces, but do not reach the other surface. Wells are used to place samples of the solution to be analyzed.

Although these test devices are used in technology, they have some disadvantages. For example, the wells of these test devices are difficult to fill. To fill each of the wells with solution samples, special dispensing systems, for example, large pipetting systems, are required. These dispensing systems are expensive and difficult to operate. Their use leads to an increase in the total cost of testing.

Another problem of these test devices is related to their design. The bottom of each well in such test plates should be transparent so that light can pass through the samples during testing. However, the remainder of the test tablet must be made of opaque material. The design of a test device with such characteristics determines its complexity and high cost.

Another disadvantage of the known test devices is that the operator determines the position of a single well in the test device. Typically, such a test device contains a large number of wells located at the same distance from each other. It is difficult for an operator to determine the position of an individual well in a large population of wells.

Summary of the invention

The basis of the present invention is the task of creating a test device for high-performance screening of improved design.

A method for holding samples, according to one embodiment of the present invention, consists in using a test plate having a pair of opposing surfaces and a plurality of channels, each of which extends from one of the opposite surfaces to the other, then at least one of the opposite surfaces of the test tablet immersed in the analyzed solution, while part of the solution penetrates into each of the channels through the corresponding holes located on the immersed surface, all kinds of gases in the channels exit the channels through the corresponding holes located on the opposite surface, the test plate is removed from the solution, and a certain amount of solution is held in each channel due to surface tension, the opposite surfaces of the test tablet are placed above the supporting surface and the solution is analyzed contained in at least one of the channels.

A method for identifying the location of at least one sample solution, according to another embodiment of the present invention, consists in using a test tablet having a pair of opposing surfaces and a plurality of channels, each of which extends from one of the opposite surfaces to the other, the channels being in a tablet, organized into groups and each group contains at least two “horizontal” rows and two “vertical” rows of channels, the solution is loaded into channels and anal ziruyut, based on the results of the analysis solution in the at least one channel is identified for further study. The location of the identified channel is marked based on the group in which the channel is found.

A method for screening a sample, according to another embodiment of the present invention, consists in preparing a sample solution for screening, using a test plate having a pair of opposing surfaces and a plurality of channels, each channel extending from one of the opposite surfaces of the test tablet to another, then at least one of the opposite surfaces of the test tablet is immersed in the solution, while part of the solution penetrates into each of the channels through the respective Twisting holes located on the submerged surface of the test tablet remove the test tablet from the solution, and at least part of the solution is held in the channels due to surface tension, analyze the solution in one or more channels.

A device for holding samples of a solution containing cells for analysis contains a test plate having a pair of opposing surfaces and a plurality of through channels, each of which extends from a hole in one of the opposite surfaces of the test tablet to a hole in the other of the opposite surfaces and is sized to accommodate the collection of cells, the area of at least one of the opposite surfaces of the test tablet on which the channels are located, is buried, for the gap between the holes of the test tablet and the corresponding of the opposite surfaces.

A device for holding samples for analysis, according to another embodiment of the invention, comprises a test plate having a pair of opposing surfaces and a plurality of channels, each of which extends from one of the opposite surfaces to the other, the channels on the test plate being combined into groups, each of which contains at least two “horizontal” rows and two “vertical” rows of channels.

A method and apparatus for holding samples for analysis according to the invention provides several advantages. For example, the present invention simplifies testing. Samples of the solution to be analyzed can be loaded into the test plate simply by recessing or immersing one of the surfaces of the test tablet in the solution. As a result, the need to use special dispensing systems to load the solution into the wells of the test plate is eliminated.

According to the invention, the design of the test device is also simplified. The only design requirement is that there is free space near one of the opposing surfaces of the test device by using additional gaskets or machining to create a recessed area and that multiple channels are drilled in the recessed area of the tablet. Unlike the known test devices, the present invention does not require any special manufacturing methods to ensure the transparency of the bottom of each well, since the channels pass through the plate through.

The present invention also facilitates the work of the operator in identifying a single channel filled with a sample for further analysis. The channels are placed on the test tablet not at equal distances, but, according to the invention, are placed in groups containing at least two “vertical” rows and two “horizontal” rows of channels, and the groups are grouped into sets of two or more groups. The distance between the groups is greater than the distance between the channels in each group, and the sets of groups are more distant from each other than the groups in each set. As a result, it is easier for the operator to identify a separate channel based on which set, in which group, in which “vertical” and “horizontal” row, the channel is located on the test plate.

Brief Description of the Drawings

The invention is further explained in the description of the preferred embodiments with reference to the accompanying drawings, in which

Figure 1 depicts a test tablet with multiple through channels (top view), according to the invention;

Figure 2 is a section along the line II-II in figure 1 according to the invention;

Figure 3 is a General perspective view of another test tablet with multiple through channels (disassembled), located between two evaporator plates, according to the invention;

4 is a block diagram of a test device comprising a test tablet with multiple through channels, according to the invention;

5 is a top view of a test tablet with multiple through channels, according to the invention;

6 is a section along the line VI-VI in figure 5 according to the invention;

7 is a top view of a test tablet with multiple through channels, according to the invention;

Fig. 8 is a plan view of an assembly of test platinum according to the invention;

Fig.9 is a General view of the assembly, in which some sections are removed, according to the invention.

Detailed description of preferred embodiments of the invention.

The test device 10 (FIG. 1), according to the invention, comprises a test plate 12 having a pair of opposing surfaces 14 and 16 (surface 16 is shown in FIG. 2) and a plurality of through channels 18. The through channels 18 are located in the recessed sections 20 and 22 of the respective sides of the test plate 12. The through holes 18 are also organized into groups 24, each of which contains at least two “vertical” rows and two “horizontal” rows of channels 18, and into sets 26, consisting of two or more groups of channels 18. The test noe device 10 provides several advantages, including simplification of sample solution loading procedure S channels 18 in the test apparatus 10, simplifying the design of the test device 10 and to facilitate the work of the operator identifying the individual channel 18 filled.

The test device 10 (figures 1 and 2) contains a test plate 12, which is made of an opaque material, for example, aluminum and polypropylene. Other materials can be used, for example, Teflon, polystyrene, stainless steel, polyethylene, any metal or plastic. The test plate 12 may also be made of transparent materials, for example, glass or transparent plastic, if the analysis is carried out by non-optical means, for example, materials blotted on membranes are analyzed.

The test plate 12 comprises a pair of opposing surfaces 14 and 16. According to the described embodiment, the opposing surfaces 14 and 16 are substantially flat, except for the recessed portions 20 and 22. The surfaces 14 and 16 may otherwise be arranged with respect to each other. Each of the opposing surfaces 14 and 16 contains one of the recessed portions 20 and 22 created by machining the test plate 12, although other methods of forming the recessed portions 20 and 22 can be used, for example, molding or adding gaskets. When any of the opposing surfaces 14 and 16 of the test plate 12 is on the supporting surface 28, the recessed portion 20 or 22, together with the multiple channels 18 located on the recessed portion 20 or 22, are separated from the supporting surface 28. If the holes 30 and 32 channels 18 were in contact with the supporting surface 28, then the solutions located in the channels 18 would have flowed from them. According to the described embodiment, on each of the opposite surfaces 14 and 16 due to the recessed sections 20 and 22, a collar 34 is formed that extends along the perimeter of the test plate 12. Although the channels 18 are separated from the supporting surface 28 by the recessed sections 20 and 22, the channels 18 can be separated from the supporting surface 28 by means of other supporting structures, for example, brackets attached to the test plate, which support the test plate 12 and the channels 18 above the supporting surface 28.

The other test device 50 shown, FIGS. 5 and 6, according to another embodiment of the present invention, is identical to the test device 10, except that the test device 50 does not contain a pair of recessed portions. Instead, the test device 50 includes a recessed portion 52 and a protruding portion 54. When the test plate 51 is located on the supporting surface, the recessed portion 52 must face the supporting surface so that the channels are separated from the supporting surface. Opposite surfaces of the test tablet 51 may have various configurations. For example, the protruding portion 54 may be flush with the upper surface of the test tablet 51.

The test plate 12 (FIGS. 1-3) also contains an auxiliary handle 36 and an opening 38 on one side for receiving one end of the handle 36. Other means of connecting the handle 36 to the test tablet 12 can be used, for example, the handle 36 can be bolted. The handle 36 protrudes from the test tablet 12 and is used to manipulate the test tablet 12 during loading and testing.

Multiple through channels 18 are formed in the test plate 12. The channels 18 extend from holes 30 located in a recessed portion 20 of one of the opposite surfaces 14 to holes 32 located in a recessed section 22 of another of the opposite surfaces 16. According to the described embodiment, the channels 18 they have an almost cylindrical shape, although they can also have a different shape, for example, a hexagonal cross section or a conical shape. Each of the channels 18 has a diameter of about one millimeter and can accommodate about 5.5 microliters of solutions of S and C cells, although the diameter, volume and number of C cells for each channel 18 can be varied as necessary or as desired. Solution S together with cells C located in solution S are retained in channels 18 due to surface tension forces, as shown in (Fig. 4). In each case, one or another size of the channels 18 may be required depending on the analyzed solution S and the surface tension properties of the solution. For example, it will be apparent to those skilled in the art that the buffer solution and culture media containing salt may differ in surface tension. Adequate surface tension is required to hold samples of solution S in channels 18.

One of the advantages of the present invention is that the test tablet 12 is easy to manufacture. A tablet having opposing surfaces may contain the required number of through channels drilled therein. A tablet may comprise one or more recessed portions 20, 22, and through channels may extend through a recessed portion of tablet 12. Because the channels 18 are through, a transparent bottom for each channel 18 is not required. During testing, light entering channels 18 will pass through them.

When using the wells, as indicated in the prior art, the test device should also be opaque, but since the holes do not pass through the device, the bottom of each of the holes must be made of transparent material so that light can penetrate the sample for optical analysis. These well-known test devices are complex and expensive.

Test plate 12 (FIG. 1) contains about two thousand channels 18 that pass through it from one surface 14 to another surface 16 opposite to it, although the number of channels 18 can be varied as necessary or as required. To make it easier for the operator to identify a single channel 18, according to the described embodiment, the channels 18 are organized into groups and sets. Each group 24 contains at least two “horizontal” rows and two “vertical” rows of channels 18, and each set 26 contains at least two “horizontal” rows and two “vertical” rows of groups 24. According to the described specific embodiment, each a group of 24 channels 18 contains five “horizontal” rows and five “vertical” rows of channels 18, and in this example there are eighty groups 24 of twenty-five channels 18, although the number can be varied as necessary or as required. The channels 18 are approximately 1.5 mm apart from each other between the “horizontal” rows of channels 18 and the “vertical” rows of channels 18 in each group 24, although this distance can be varied, and the gap between the “horizontal” rows of channels 18 may differ from the gap between the “vertical” rows of channels 18 in each group 24. According to the described embodiment, each set of groups 24 contains two “horizontal” rows of groups 24 and ten “vertical” rows of groups 24, and thus there are four sets 26, each of which contains dv there are eleven groups of 24 channels 18, although the number can vary. Groups 24 included in set 26 are spaced about 2.0 mm apart, and sets 26 of groups 24 of channels 18 are spaced about 2.5 mm apart, although these distances can be varied as needed or as desired.

By arranging the channels 18 into sets 26 and groups 24, it is much easier for the operator to identify a separate channel 18 in the test plate 12 and to search for a separate sample. With the help of sets of 26 channels 18, the operator can navigate in which area channel 18 is located, and with the help of groups 24 the operator can more accurately determine the location of channel 18. The “vertical” and “horizontal” row of channel 18 in each group 24 defines the exact location of channel 18 The gaps between sets 26, groups 24 and “vertical” and “horizontal” rows are different from each other, so that it is easier for the operator to visually identify a separate channel 18. When all channels 18 are at the same distance from each other, tr it is more difficult to identify a separate channel 18, and the operator can easily confuse the channel and select a sample from the wrong channel 18.

Although the channels 18 in the test devices 10 and 50 are organized into groups 24 and sets 26, other channels 18 can be used. For example, when the test device is controlled by a robot and not an operator, the channels 18 can be located at equal distances from each other, as shown in another embodiment of the test device 60 (FIG. 7).

The test device 60 is identical to the test devices 10 and 50, except that the channels 18 are at the same distance.

Test device 10 (FIG. 3) may also comprise a pair of auxiliary evaporation plates 40 and 42. Evaporation plates 40 and 42 are attached to respective opposed surfaces 14 and 16 of test plate 10. Evaporative plates 40 and 42 are attached to test plate 12 with bolts, clamps, or other mechanical means. When the evaporation plates 40 and 42 are attached to the test plate 12 over the recessed sections 20 and 22, due to the presence of the recessed sections 20 and 22 on the opposite surfaces 14 and 16 of the test plate 12, the openings 30 and 32 of the through channels 18 remain at some distance from the evaporation plates 40 and 42. Evaporation plates 40 and 42 help protect samples of solution S located in the channels 18 of the test plate 12 from evaporation and contamination.

Instead of a plate 12 with a recessed portion, an assembly can be proposed in which gaskets are located between the test plate and the evaporation plates, whereby the openings of the through channels are separated from the evaporation plates. In addition to the gaskets between the test plate and the evaporation plates or instead of them, recessed areas can be provided in the evaporation plates. To provide a gap between the openings of the through channels and the evaporation plates, you can use the recessed areas on the test plate, the buried areas on the evaporation plates or gaskets in any combination.

According to an embodiment of the present invention, a stack of test plates is provided in which evaporation plates can further be arranged between the test tablets. Test plates included in the stack may have recessed portions, or recessed portions may be provided in the evaporation plates located between the test plates. To ensure that each test plate in the stack of test plates is separated from the surface of an adjacent test tablet, evaporator plate, or both together, recessed portions in the test tablets, recessed portions in the evaporation plates, or gaskets in any combination can be used.

Consider one example of the use of the claimed invention.

In this example, C cells are mutagenized by ultraviolet radiation, chemical mutagenesis, or other mutagenesis technology. Cells are grown with the possibility of separation. Having grown C cells, they are diluted to a concentration of one C cell per ten microliters in a medium containing a fluorescence inducing substrate or a chromogenic substrate. For this example, solution S is considered as a medium with cells C. As a result, the cells are randomly distributed along channels 18, and many channels 18 contain one or more C.

Although one example of the preparation of a solution of S with cells C has been considered, it will be apparent to those skilled in the art that other methods and techniques for preparing samples to be tested using test device 10 can be used (FIGS. 1-4).

A test plate 12 is used comprising a pair of opposing surfaces 14 and 16 and channels 18 that extend from one of the opposite surfaces 14 to another of the opposing surfaces 16. At least one of the opposite surfaces 14 of the test tablet is immersed in the prepared solution S. Solution S penetrates into the openings 30 and 32 of each of the channels 18 of the test plate 12, and various gases in the channels 18 can escape through the openings 30 and 32 at the opposite end of the channels 18. Alternatively, test The plate 12 can be immersed in solution S so that solution S enters through the upper hole 30 of each channel 18.

One of the advantages of the present invention is the simplicity of loading solution S into each of the channels 18. According to the above description, it is possible to load samples of solution S into all channels 18 of the test plate 12 within a relatively short period of time and without the use of any specialized solution distribution system. Known test devices equipped with wells require the use of a special solution dispensing system, for example, bulky pipetting devices, by means of which a solution is loaded into each of the holes. These specialized solution dispensing systems are difficult to use and expensive.

After the solution S has been drawn into the channels 18, the test plate 12 is removed from the solution S. The surface tension holds the solution S in each of the channels 18. According to the described embodiment, each channel 18 has a diameter of about one millimeter and holds about 5.5 microliters of solution S with cells C (figure 4), although the diameter and volume of each channel 18 can be varied as necessary or as desired depending on the specific application. The handle 36 can be used to control the position of the test tablet 12 during the above operations.

After removing the test plate 12 from the solution S, it is placed on the supporting surface 28. Since the channels 18 are located in the recessed portion 22 of the test tablet 12, the openings 32 of the channels 18 are separated from the supporting surface 28, therefore, the solution S held by surface tension forces remains in the channels 18 A pair of evaporation plates 40 and 42 can be attached to opposite surfaces 14 and 16 of test plate 12, which protect samples of solution S in the test plate from evaporation and contamination.

Test plate 12 can then be incubated, maintaining a temperature of about 37 ° C and a humidity of about 70%, although temperature and humidity can vary depending on the particular application. During the incubation process, the cells multiply and produce the desired protein (the cells can produce an enzyme, antibody, or metabolite to be studied). Analyze the ability of a protein, for example, an enzyme, to hydrolyze a substrate, for example, by measuring fluorescence inducing groups or chromogenic groups released by hydrolysis.

Although one example of processing samples of a solution of S in a test plate 12 is considered, other methods for processing and analyzing samples known to those skilled in the art can be used.

Then, samples of solution S with cells C located in channels 18 are tested using an image analyzer equipped with a light source 44 and a detector 46. Light propagates from the light source 44 towards the openings 30 of channels 18 of the test plate 12 and through solution S located in the channels 18 of the test plate 12. The detector 46 is located on the opposite side of the test plate 12 and detects the light transmitted through the solution S located in the channels 18. Based on the differences between the registered light and emitted light can get information about the characteristics of the individual samples S solution methods known to those skilled in the art. The image analyzer is able to determine which channels 18 contain solution 3 with the highest concentration of the converted substrate and, therefore, the largest amount of enzyme. The task in this case is to identify C cells producing the largest amount of enzyme. Similarly, you can identify the cells that secrete the highest amount of the desired protein or chemical.

Although an example of optical analysis of samples of solution S in a test plate 12 is considered, other methods for analyzing samples can be used, in particular non-optical. For example, a plate containing samples of a solution of S with cells C can be blotted onto a membrane and used to perform Western blot analysis or, alternatively, samples S with cells C can be blotted onto a substrate containing material, which makes it possible to visually measure the modification of the substrate. As a result, when non-optical means are used to analyze the solution samples in the test plate 12, the test plate 12 can be made of a transparent material.

Then the operator searches for samples of solution S with the highest concentration of the converted substrate. The channels 18 in which the solution S with the highest concentration of the converted substrate is located can be identified on the basis of which set of 26 groups 24, in which group 24 and in which “horizontal” and “vertical” row in each group 24 each identifiable channel is located 18. One of the advantages of the present invention is that the channels 18 are organized into groups 24 and sets 26, which allows the operator to easily identify a separate channel 18 on the test plate 12. After finding the right samples, the operator can test Further analysis of these samples by known methods.

You can use other methods and examples of finding samples. For example, if a robot is used to locate and locate a single sample, another test device 60 may be used (FIG. 7). The robot does not need channels 18 to be organized into groups 24 and sets of 26 channels 18, although such an organization can even help the robot identify and retrieve the desired patterns.

According to embodiments of the present invention, the test tablet is in the form of an assembly or base. For example, a tablet may comprise a plurality of individual components that together form an assembly having opposing surfaces and a plurality of through channels extending from one surface to another. Figures 8 and 9 show an example embodiment of the present invention when the test tablet containing the assembly is a tablet made of a bundle of capillary tubes.

The tablet, base or assembly 70 comprises a bundle of capillary tubes 72 connected together by a brace 74. The through channels of the assembly, according to this embodiment, are channels that extend longitudinally through the center of each capillary tube. The bandage 74 may have opposing surfaces 76 and 78 that are substantially flat and parallel to each other. The bandage may be made of metal, plastic, glass, rubber, elastomeric or any other suitable material. Each capillary tube 72 has a first end 80 and a second end 82. The first ends 80 of the capillary tubes form the surface 84 of the base or assembly 70, and the second ends 82 of the capillary tubes 72 form the surface 86 of the base or assembly.

Each bundle capillary tube 72 constituting the base or assembly 70 has a length, measured from its first end 80 to its second end 82, which is at least two times the average diameter of each tube. Preferably, the length of each tube is more than four times the average diameter of each tube and it is preferable that it is many times greater than the average diameter. Each capillary tube may be, for example, a microcapillary tube or a hollow fiber optic fiber.

Capillary tubes may be in the form of a hollow cylinder or rounded, oval or polygonal cross sections.

The average diameter of each capillary tube is preferably from about 0.001 mm to about 1 mm, and the length of each tube is preferably from about 1 mm to about 1 cm. The dimensions of the capillary tubes are preferably such that each tube can hold from about 0.0001 microliters to about 10 microliters. a liquid sample, for example, 5.5 microliters, although the diameters, lengths and capacities of capillary tubes can be varied as needed or as desired. According to some embodiments of the invention, it is not necessary to use a bandage to jointly hold the capillary tubes in the bundle, since the tubes can be fused or bonded, glued together to be held together in the bundle.

The number of capillary tubes is preferably from about 100 to about 1000 or more, capillary tubes, for example, from about 500 to about 1500. It is preferred that the tubes are arranged in “horizontal” rows and the rows are arranged to form “vertical” rows. Although the bundle of capillary tubes 72 (FIGS. 8, 9) has a circular cross section and the brace 74 is ring-shaped, the scope of the present invention also provides other shapes of the bundle and brace. For example, a rectangular or square array of capillary tubes surrounded by a band can be provided, and the band in this case also preferably has a rectangular or square shape. Due to the rectangular or square shape of the arrays of capillary tubes, the various “horizontal” rows and “vertical” rows of capillary tubes can be easily identified, which facilitates the identification of one capillary tube in the array.

The bandage 74 surrounding the bundle of capillary tubes has a length measured between opposite surfaces 76 and 78, which exceeds the length measured between opposite ends 80 and 82 of the capillary tubes. Due to this, the associated assembly can be placed on the surface of, for example, an analytical instrument and the ends of the capillary tubes will not touch the surface. In addition, assemblies can be stacked without affecting the surface tension forces acting in the through channels.

The assembly (FIGS. 8 and 9), like the tablets (FIGS. 1-7), can be loaded or filled with the original liquid sample to obtain a plurality of samples, each of which constitutes a portion of the initial liquid sample. Alternatively, the assembly can be loaded with several source liquid samples, with each source liquid sample filling at least one of the through channels. In this case, the words “download” or “fill” mean that they fill at least partially, but not necessarily completely. The through channels can be loaded or filled, for example, by immersing the assembly or tablet in a liquid sample, bringing into contact at least one of the opposing surfaces of the assembly or tablet with a liquid sample, or by bringing the inner walls of the respective through channels into contact with the liquid sample or with the corresponding liquid samples .

The contact of a liquid sample with one of the opposite surfaces can be carried out by immersion, heating, pipetting, dripping or otherwise, loading or at least partially filling the set of capillary tubes or through channels so that, due to the capillary effect, portions of the liquid sample are drawn into the corresponding capillary tubes or through channels. After removing or breaking the contact of the liquid sample with the assembly or tablet, the opposite surfaces of the assembly or tablet are preferably cleaned of the liquid sample so that portions of the sample remaining inside the respective capillary tubes are isolated from each other.

Automated filling devices can be used, and they are preferred if it is important that the corresponding liquid samples or portions of the liquid sample are in contact only with the inner walls of the through channels and that contact with opposing assembly surfaces is to be avoided.

According to other embodiments of the invention, a method for high throughput screening is provided. The method allows you to screen at least one liquid sample that contains the target component or substance to be analyzed. In this case, the target component or substance to be analyzed is called “analyte”. An analyte may be, although not necessarily, a biological sample. An analyte exhibits a detectable property or exhibits a detectable characteristic in the presence of an indicator or a similar substance or as a result of reaction with it. For example, an analyte may independently exhibit a fluorescence property. After a liquid sample has at least partially filled a plurality of through channels, portions of a liquid sample containing an analyte can be detected and it is determined which of the through channels contains a portion of a sample exhibiting fluorescence property.

In another example, the analyte itself does not exhibit a detectable property, but instead can cause the indicator to manifest itself in the indicator as a result of a reaction with it. According to such an embodiment, the through channels of the test assembly can be preloaded or subsequently loaded with one or more indicators, as a result of which, after loading the liquid sample into the set of through channels, portions of the sample containing analyte can react with the indicator and, thus, determine the manifestation of the detectable properties indicator. In this case, although the analyte itself does not exhibit a detectable property, indirect detection of the analyte is possible, since the presence of the analyte determines the detectable property of the indicator, which, in turn, can be detected directly. Moreover, the methods of the present invention provide for the separation and isolation of analytes from the initial liquid sample.

According to a high throughput screening method, portions of a liquid sample are loaded into a test assembly having a pair of opposing surfaces and a plurality of through channels, each of the through channels extending from one of the opposite surfaces to the other. Download leads to at least partial filling of the set of through channels with portions of the liquid sample, and surface tension holds the corresponding portions in the corresponding set of through channels. Alternatively, multiple liquid samples may be loaded into respective through passages. The method also includes detecting which portion of the sample in the through channels exhibits a detectable property.

According to embodiments of the present invention, the assembly for high throughput screening preferably comprises at least about 100 through channels, more preferably at least about 500 through channels and, according to some embodiments of the present invention, up to 1,000,000 through channels. In high-throughput screening, you can use the specified device, which allows you to analyze more than 100000000 samples or portions of the sample per day using a single assembly.

The analyte to be screened may be, for example, a biological cell, a mixture of biological cells, a mutant cell, a secreted protein, an enzyme, a microorganism, a mixture of microorganisms, contamination, or combinations thereof. An analyte may be a population of random mutants of one or more organisms. If the analyte is a mixture of biological cells, then it may be a random sample isolated from the natural environment.

 A detectable property may be, for example, a fluorescence or absorption property. Before filling a high-performance assembly, a liquid sample can be diluted with a suitable diluent to obtain an analyte concentration in a liquid sample such that, as a result of filling a plurality of through channels with a sample, the fraction of through channels where at least one of the analytes got into is from a quarter to half through channels.

In some cases, an organism with the desired properties can be identified, even if the organism has entered through holes in a mixture with other organisms. Under such conditions, a mixture of other organisms, for example, a mixture of biological cells, can be diluted before filling so that several organisms or cells fall into each through channel. Using this dilution method, the presence of an analyte can be detected. For example, one particular mutant can be found in a cluster of many biological cells and their mutants instead of having many cells in the mixture present in each channel. Thus, for example, if a sample contains 1,000,000 cells and only one of them is a target mutant cell, which we call an “analyte”, and a test plate with 10,000 through channels is used, then the sample can be diluted so that 1,000,000 cells fall into the through channels with portions of the sample, and each portion contained about 100 cells. In cases where the analyte characteristic is detectable, despite the presence of many other cells in the same through channel, it is possible to isolate the analyte from 99.99% of the sample in one analysis.

Test tablets used in accordance with the present invention, including the tablets shown in FIGS. 1-1 and the assemblies shown in FIGS. 8 and 9, may contain hydrophilic materials or coatings, hydrophobic materials or coatings, or a combination thereof to facilitate loading portions of the liquid sample into the through channels. For example, opposing surfaces of the assembly can be made of hydrophobic material or processed so that liquid samples are expelled from the surface with the exception of areas directly adjacent to the openings of the through channels on one of the opposite surfaces. According to such an embodiment, portions of the liquid sample can be drawn into the through channels due to the capillary effect without wetting the opposite surfaces of the tablet. As a result, when the tablet is loaded and separated from the liquid sample, no liquid connection occurs between the individual through channels and contamination of the divided portions of the sample is minimized. According to some embodiments of the present invention, the inner walls of the through channels can be made of a hydrophilic material or coated with it, since this material can be easily wetted by an aqueous sample or aqueous medium. The inner walls of the through channels can be completely made of hydrophilic material or processed by it, and can be partially made of such material or processed by it. Tablets having hydrophilic inner walls of the through channels and hydrophobic opposite surfaces are a good means of retaining, isolating, or restricting the position of liquid samples in the through channels of the test tablet and at the same time keep adjacent sections of opposite surfaces practically free of the liquid sample.

According to some embodiments of the present invention, in order to promote a capillary effect, it is desirable to place the hydrophilic material in close proximity to the hole of each through channel on one of the opposite sides, while maintaining or providing hydrophobia or lack of hydrophilia in the remaining region of the same surface. One of the opposite surfaces of the test tablet or both of them can be made of hydrophobic, hydrophilic or both materials or processed by them, as discussed above, although if the through channels are to be loaded by immersion, it is preferable that one of the opposite surfaces that comes into contact with liquid the sample was treated with hydrophobic material or made of it with the exception of areas directly adjacent to the holes of the through channels on this surface and preferably around lev els of them.

High-throughput screening methods that can be used with the assemblies and other plates of the present invention allow transcription absorption analysis, fluorescence transcription analysis, fluorescence analysis of secreted enzymes and screening analysis of microorganisms. These and other suitable assays for which the use of tablets and methods of the present invention may be useful are described, for example, in Arndt et al. "A rapid genetic screening system for Identifying gene-specific suppression constructs for use in human cells", Nucleic Acids Res., 28 (6): E15 (2000), or Rolls et al. "A visual screen of a GFP-fusion library Identifies a new type of nuclear envelope membrane protein", J. Cell Boil, 146 (1): 29-44 (1999), or Sieweke, "Detection of transcription factor partners with a yeast one hybrid screen". Methods Mol. Biol., 130: 59-77 (2000); and WO 97/37036.

Claims (22)

1. Test plate for holding samples for analysis, containing a plate having a pair of opposing surfaces, a plurality of channels in the plate, each of which extends from one of the opposite surfaces to the other and has a constant diameter from one surface to the other, the channels are combined into groups, each of which contains at least two horizontal and two vertical rows of channels, characterized in that each of the channels has a hydrophilic inner wall, a diameter of about 1 mm or less, and holds from 0.1 to 10 μl of the solution. 2. The tablet according to claim 1, characterized in that on at least one of the opposite surfaces of the fixed evaporation plate. 3. The tablet according to claim 1, characterized in that it further comprises a patch intended to cover at least one surface of the plate, release means for providing a gap between the patch and the plurality of through holes. 4. The tablet according to claim 3, characterized in that the release agent comprises a gasket. 5. The tablet according to claim 3, characterized in that it further comprises a recessed portion where the through channels are located, wherein the release agent is a shallow portion of the plate. 6. The tablet according to claim 1, characterized in that it further comprises sections for doses of a liquid sample held by surface tension in a corresponding plurality of channels. 7. The tablet according to claim 1, characterized in that the through channels are cylindrical in shape. 8. A system comprising a plurality of test tablets according to claim 1, wherein said test tablets are stacked one above the other. 9. The analysis method, including the use of a test assembly containing a pair of opposing surfaces and a plurality of through channels in the plate, each of which extends from one of the opposing surfaces to the other, has a hydrophilic inner wall and a constant diameter from one surface to another opposite surface of about 1 mm or less, at least partially filling a plurality of through channels with a plurality of doses of a corresponding liquid sample while retaining them in the channels due to the surface tension analysis and analysis of at least one of the liquid samples in at least one through channel by absorption or fluorescence transcription analysis, fluorescence analysis of secreted secreted enzymes and screening analysis of microorganisms, said through channels being combined into groups, and each group contains at least two horizontal and two vertical rows of through channels, at least one of the doses of the corresponding liquid sample contains an analyte that directly or indirectly showed a detectable response in one or more assays, wherein further comprising determining which of a plurality of through holes comprises a dose of a liquid sample, comprising said analyte. 10. The method according to claim 9, characterized in that at the partial filling step, the test assembly is immersed in a liquid sample. 11. The method according to claim 9, characterized in that it is additionally placed in a stack of one test assembly on top of another test assembly. 12. The method according to claim 9, characterized in that when carrying out one or more of the first dose analysis of a liquid sample in an appropriate set of through channels, the sample is tested using optical means. 13. The method according to claim 9, characterized in that the dose is interacted in the corresponding set of through holes until the determination in which of the specified set of through holes contains the dose of the liquid sample in which the analyte is located. 14. The method according to claim 9, characterized in that after analyzing the liquid samples, the test assembly is incubated at a given temperature. 15. The method according to claim 9, characterized in that the dose of the liquid sample is transferred to the membrane and the membrane is analyzed to determine the desired properties of the doses of the liquid sample. 16. The method according to clause 15, wherein the analysis of the membrane includes a visual analysis. 17. The method according to clause 15, wherein the analysis of the membrane includes Western transfer (detection of slots). 18. The method according to clause 15, wherein introducing a group of cells into the medium in the through holes of the test assembly, incubate the test assembly under conditions suitable for cell group growth, measure the properties of a group of cells in a test assembly, at least one group of cells is selected based on the desired measured properties. 19. The method according to p. 18, characterized in that to measure the properties of the group of cells in the test assembly measure the activity of the expressed protein. 20. The method according to p. 18, characterized in that when the group of cells is introduced into the medium in the through holes of the test assembly, a plurality of cells are delivered to each of the through holes of the set. 21. The method according to p. 18, characterized in that when the group of cells is introduced into the medium in the through holes of the test assembly, the reagent is additionally delivered together with a plurality of cells introduced into the through hole matrix. 22. The method according to p. 18, characterized in that when measuring the properties of a group of cells in a test assembly, testing a plurality of cells with an optical means.

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