RU106378U1 - DEVICE FOR ANALYSIS WITH APPLICATION OF THE BIOCHIP - Google Patents

Abstract

 1. A device for conducting analysis using a biochip, comprising a housing in which a circuit consisting of channels for supplying and discharging liquid and a compartment is placed, in which a removable cell for placing the biochip, having locking devices, and equipped with a hermetically sealed removable cover, is hermetically fixed. ! 2. The device according to claim 1, characterized in that it is equipped with a device locking the circuit installed in the device body. ! 3. The device according to claim 1, characterized in that the container for accommodating the biochip has a removable hermetically sealed bottom. ! 4. The device according to claims 1 and 4, characterized in that the bottom of the tank for accommodating the biochip has an adhesive coating. ! 5. The device according to claim 1, characterized in that it is equipped with a removable device, hermetically mounted on the surface of the biochip, and dividing the container in which the biochip is placed, into several containers of smaller volume, the bottom of each of which is formed by the surface area of the biochip. ! 6. The device according to claim 1, characterized in that it is equipped with a tank for washing liquid. ! 7. The device according to claim 1, characterized in that it is equipped with a container for collecting waste fluid. ! 8. The device according to claim 1, characterized in that it is equipped with a device for measuring fluid pressure. ! 9. The device according to claim 1, characterized in that it is equipped with a device for trapping air bubbles. ! 10. The device according to claim 1, characterized in that it is equipped with a device pressing the removable cover. ! 11. The device according to claim 1, characterized in that it is equipped with a device for additional introduction of fluid into the container in which the biochip is placed. ! 12. The device according to claim 1, characterized

Description

The utility model relates to the field of medicine, and can also be used in the field of biology and veterinary medicine and is intended for research using biochips of cells (or other biological objects with microscopic dimensions).

A device is known which is a flow chamber designed to incubate a biochip with a cell suspension and to wash it, containing a lead-in and outlet-tube, as well as a housing in which a capillary is located, one of the walls of which forms a biochip (see A.V. Shishkin, I.I. Shmyrev, S. A. Kuznetsova, N. G. Ovchinina, A. A. Butylin, F. I. Ataullakhanov, A. I. Vorobyov “Immunological biochips for the study of human red blood cells”, the journal “Biological membranes”. - 2008.- No. 4 - S.267-276), taken as an analogue.

The disadvantage of this device is the high complexity of the preparatory stages of the analysis using a biochip, including the processing of the biochip with a solution of a substance that blocks the sites of non-specific binding of the substrate and the subsequent washing of the biochip with a detergent solution. Appropriate solutions require sequentially filling the entire circuit of the device (capillary and flow tubes), which leads to a significant consumption. In this case, it is necessary to monitor the tightness of the system and avoid air bubbles entering the circuit, which also increases the complexity. In addition, the disadvantage is the not entirely successful fastening of the biochip in the device, requiring precise manipulations when fixing the biochip in it and its subsequent removal. This increases the risk of damage to the biochip. The design of the flow chamber allows additional introduction of small volumes of reagent solutions into its capillary. But at the same time, the entire surface of the biochip is brought into contact with these solutions. This leads to a fairly high consumption. There is no possibility of simultaneous contacting with different solutions of individual sections of the surface of the biochip.

A device for incubation and washing of a biochip designed to detect surface antigens of cells is known, used to incubate a biochip with a cell suspension, to wash it, and also to process a biochip with various liquids during additional studies of the cells bound to it, taken as a prototype and representing a container (for example, a Petri dish), at the bottom of which a biochip is fixed. (See, for example, A.V. Shishkin, I.I. Shmyrev, S. A. Kuznetsova, N. G. Ovchinina, A. A. Butylin, F. I. Ataullakhanov, A. I. Vorobyov “Immunological biochips for the parallel determination of surface antigens and morphological studies of cells ”, Biological membranes, 2008, volume 25, No. 4, pp. 277-284).

The disadvantage of this device is the inability to adjust the flow rate of the washing liquid. This leads to the fact that when washing the biochip from non-specifically bound cells, separation of those cells that specifically bound to the biochip, but with insufficient strength, can occur. As a result, the sensitivity of the analysis and reproducibility of the results are reduced. In addition, the fastening of the biochip and its extraction require precise manipulations. This poses a risk of damage to the biochip. The disadvantage is the impossibility of simultaneously bringing individual sections of the surface of the biochip into contact with different solutions without mixing them.

The objective of the claimed utility model is to reduce the complexity of the analysis, increase the reproducibility of the results and expand the functionality of the device.

The problem is solved due to the fact that, according to a utility model, a device for analysis using a biochip, containing a container for placing a biochip, is equipped with a platform, a locking device, a hermetically sealed removable cover, channels for supplying and discharging liquid.

The capacity for placing the biochip is removable and equipped with a locking device.

The platform is equipped with a compartment in which a removable container is tightly fastened to accommodate the biochip.

The capacity for placing the biochip has a removable hermetically sealed bottom.

The bottom of the biochip container has an adhesive coating.

The device is equipped with a removable device, hermetically mounted on the surface of the biochip, and dividing the container in which the biochip is placed, into several containers of smaller volume, the bottom of each of which is formed by the surface area of the biochip.

The device is equipped with a tank for washing liquid.

The device is equipped with a container for collecting waste fluid.

The device is equipped with a device for measuring fluid pressure.

The device is equipped with a device for trapping air bubbles.

The device is equipped with a device pressing the removable cover.

The device is equipped with a device for additional introduction of fluid into the container in which the biochip is placed.

The device is equipped with a drainage tube.

The shape and dimensions of the horizontal section of the tank for placing the biochip correspond to the shape and dimensions of the biochip.

Using the claimed utility model allows to reduce the complexity of the analysis and to reduce the consumption of used solutions. It becomes possible to carry out individual stages of the analysis with filling the container with solutions and removing them in the least labor-intensive way (without regulating the flow rate of the liquid) in the same way as when working with a cuvette. Other stages of the analysis can be performed with a fluid flow at a precisely defined speed, as when working with a flow chamber. Due to this, the complexity of the analysis is reduced in comparison with the use of a device taken as an analog, while maintaining the same sensitivity and reproducibility of the results. At the same time, when using the claimed device, the sensitivity and reproducibility are significantly increased compared to using the device taken as a prototype.

The implementation of the capacity to accommodate a removable biochip increases the usability of the device. If necessary, a removable container with closed locking devices (taps) can be used separately as an incubation-washing cuvette. This increases the versatility of use of the device.

The implementation of the bottom of the tank (bottom wall of the tank) removable facilitates the extraction of the biochip from it. The presence of an adhesive coating on the bottom of the tank simplifies the process of attaching the biochip, as well as ensures the tightness of the connection of the biochip with its surface.

The presence of devices for measuring fluid pressure allows you to determine the difference between atmospheric pressure and fluid pressure in the circuit of the device. These data are necessary for estimating (calculating) the velocity of a fluid.

The presence of the device pressing the removable cover provides reliable sealing of the device.

Correspondence of the shape and size of the horizontal section of the cavity of the container to the shape and size of the biochip provides the most economical expenditure of the studied cell suspension, since all cells are deposited on the surface of the biochip during incubation.

The presence of a drainage tube equipped with a locking device makes it possible to drain the liquid from the tank without removing liquid from the higher sections of the device. In addition, the drainage tube can be used to introduce small volumes of liquid into the device circuit, as well as to remove air bubbles.

The presence of a device for trapping air bubbles prevents their penetration into a closed container when washing the biochip.

The platform provides a convenient arrangement of the device on the microscope stage. In addition, the main parts of the device are located in it.

The presence of a removable device, hermetically mounted on the surface of the biochip and dividing the container (in which the biochip is placed) into several containers of smaller volume, the bottom of each of which is formed by the surface sections of the biochip, makes it possible to simultaneously contact different parts of the surface of the biochip with solutions of different reagents without mixing them together. This allows one to reduce the consumption of solutions of expensive reagents during additional studies of cells bound to the biochip (for example, to reduce the consumption of labeled antibodies in determining coexpression of antigens).

The utility model is illustrated in the drawing, where:

figure 1 presents a General view of the device in longitudinal section;

figure 2 presents a top view of the device;

figure 3 presents a section AA in figure 2;

figure 4 presents the biochip of the 1st type;

figure 5 presents the biochip of the 2nd type.

The device comprises a housing (platform) 1 (Fig. 1) in which there is a driving channel 2, a discharge channel 3, a locking device (crane) 4 and a compartment 5 for accommodating a removable cell 6, in which the biochip is located 7. Tightness of the cell 6 in the compartment 5 provides a sealing coating 8. The removable cuvette 6 has a casing 9 made in the form of a frame to which a removable bottom 10 is attached. Sealing of the fastening of the bottom 10 to the casing 9 of the cuvette 6 provides a sealing coating (not shown). In addition, sealing the fastening of the bottom of the container to the casing 9 of the cuvette 6 is provided by a strip of transparent adhesive tape (not shown), which is glued to the lower wall of the cuvette 6. The bottom 10 is made of a transparent material and has an adhesive transparent coating (not shown), providing fastening the biochip 7. When attaching the bottom 10 to the body 9 of the cuvette 6, a container 11 is formed to accommodate the biochip. Channels 2 and 3, the tank 11, as well as the lead and outlet tubes (26 and 27) form the circuit of the device. The contour of the device has a shape that provides a laminar flow of fluid. The housing (platform) 1 is made of transparent material. This allows you to determine the presence of air bubbles in the circuit of the device when it is filled with liquid.

Locking devices (taps) 12 and 13 are placed in the housing 9 of the removable cuvette 6. Moreover, the cross section of their openings 14 corresponds to the cross section of the supply and outlet channels 2 and 3. In taps 12 and 13, additional openings 15 are made, communicating with the hole 14. B the ends of the taps 12 and 13 are placed elastic plugs 16 (figure 2) through which using a syringe it is possible to introduce small volumes of liquid into the circuit of the device (the needle of the syringe is drawn through the plug 16). The device is equipped with a compartment 17 designed to remove air bubbles from its circuit, filled with liquid. The device is equipped with an additional tube 18 opening in its circuit. The tube 18 is equipped with a locking device (valve or clamp), which is not shown in the drawing.

The cuvette 6 (Fig. 3) is provided with a removable cover 19. The cover 19 consists of a metal frame and a transparent plate 20. Sealing the connection of the cover 19 and the casing 9 of the cuvette 6 provides a sealing coating 21.

The device is equipped with a movable frame 22, pressing the cover 19 to the casing 9 of the cuvette 6, installed in the compartment 5. The pressing frame 22 is movably fixed in the grooves 23 of the casing (platform) 1. In the casing 1 in the projection of the location of the biochip 7 there is a hole 24 for light passage .

The shape and dimensions of the horizontal section of the tank 11 correspond to the shape and dimensions of the biochip 7.

The supply channel 2 (FIGS. 1 and 2) and the discharge channel 3 are connected by means of nozzles 25 to the supply and discharge pipes 26 and 27. The supply pipe 26 is connected to a container in which the washing liquid is located (not shown in the drawing). The discharge tube 27 is connected to a drain tank designed to collect the spent washing liquid (not shown in the drawing).

For the operation of the device, the level of the washing liquid located in the tank (not shown in the drawing) connected to the drive tube 26 must be higher than the surface of the biochip 7. A drain tank (also not shown in the drawing) connected to the discharge tube 27 is below the biochip level. The external opening of the discharge channel 3 (Fig. 1) is located below the surface level of the biochip 7, and the external opening of the driving channel 2 is located above the level of the biochip 7.

The washing liquid storage tank and the drain tank are mounted on external tripods, but other installation options are possible. The device during the controlled washing of the biochip is placed on a microscope stage.

The washing liquid container (not shown in the drawing) is equipped with a scale (not shown in the drawing) indicating the height of the liquid level above the surface of the biochip 7. It is also possible to design a device additionally equipped with a vertical water meter tube with a scale (not shown in the drawing), or other device, allowing to evaluate the fluid pressure in the circuit.

It is possible to use the device when the washing liquid does not flow into the device loop by gravity, but is supplied at a given speed due to the operation of the pump (not shown in the drawing), to which the drive tube 26 is connected (if the pump is operating in the fluid injection mode) or the discharge tube 27 (if the pump is in liquid suction mode). In this case, the height of the liquid level in the external tank does not matter.

Biochip 7 (Figs. 4, 5) is a substrate 28 on which the working region 29 is located, containing test sections 30 in which antibody molecules specific to cell surface antigens (or other biological microorganisms) are immobilized, or molecules of other substances, capable of binding to molecules located on the surface of the studied cells (or other micro-objects of biological origin). Test sections 30 are separated from each other by background sections 31 that do not contain these molecules. The working area 29 is limited by the edges 32 of the substrate 28. The device allows the study using biochips 7 with a different type of location of the test sites.

The biochip of the 1st type (Fig. 4) has test sections 30 made in the form of an arrangement of circular or other shaped spots that do not cross the working area 29 of the biochip 7 in any direction.

On the biochip of the 2nd type (figure 5), the test sections 30 are made in the form of parallel stripes crossing the work area 29

To enable some additional studies of bound cells when working with type 2 biochips, the device is equipped with a device 33 (Fig. 3), which allows dividing the container 11 into several containers (channels) 34 having smaller sizes. The bottom of each of the containers 34 is formed by the surface area of the biochip 7. The device 33 is a plate 35 having protrusions 36 provided with a sealing coating (not indicated in the drawing). The device 33 is placed in such a way that the containers 34 are located parallel to the longitudinal axis of the contour of the device, but perpendicular to the test sections 30 of the biochip of the 2nd type. (A biochip of the 2nd type is installed in the tank 11 so that its test sections 30 are located across the longitudinal axis of the device’s contour). In this case, the containers (channels) 34 communicate with other elements of the device circuit with open taps 12 and 13. To fill the containers 34 with reagent solutions, holes 37 are made in the plate 35 (Fig. 1), in each of which a Hamilton syringe needle or tip can be inserted micropipettes, or other similar devices.

To access the holes made at the ends of the taps 12 (FIG. 2) and 13 and closed with elastic plugs 16 in the housing 1, holes 38 and 39 are made in their projection. The cranes 12 and 13 are rotated using a screwdriver or a special key. To do this, recesses 40 are made at their opposite ends. For access to them, openings 41 and 42 are made in the housing 1.

The device is used as follows.

The biochip 7 is fixed in a removable cuvette 6, pressing it to the bottom 10 provided with a transparent adhesive coating. Locking devices (taps) 12 and 13 are closed. Fill the tank 11 with the necessary liquids, with which the biochip 7 is incubated. After the biochip 7 has been processed, the used solution is drained, and the tank 11 is filled with the following solution. The sequence of processing the biochip 7, designed to determine the surface antigens of cells, can be as follows: 1) processing the biochip 7 with a protein solution (for example, bovine serum albumin solution) blocking the sites of non-specific binding of the substrate 28; 2) processing the biochip 7 with a detergent solution to eliminate 28 protein molecules not bound to the substrate; 3) washing the biochip with 7 buffer solution to eliminate traces of detergent. When carrying out these manipulations, the cuvette 6 is mounted on a shaker.

A drive tube 26 and a discharge tube 27 are attached to the nozzles 25 of the device, which are connected respectively to a tank filled with a washing liquid (not shown in the drawing) and a drain tank (not shown in the drawing). Install a removable cuvette 6 in the compartment 5 of the device. Install the cover 19 and fix it using a movable pressing frame 22. Open the taps 4, 12, 13 while the liquid fills the circuit of the device (drive channel 2, discharge channel 3, tank 11). After that, the valve 12 is closed. The air from the device circuit is removed using a syringe, which is connected to the tube 18. The remaining air accumulates in the compartment 17 and can be removed using a syringe, the needle of which is inserted through a hole closed in its upper part with an elastic stopper.

The tap 13 is opened. The device is tilted so that the closed tap 12 is at the bottom. Using a syringe, a needle is inserted through an opening in the end of the valve closed by an elastic stopper 16 and the required volume of the cell suspension is introduced, which enters the container 11 through the hole 15. The valve 13 is closed. Other options for manipulating the container 11 with a cell suspension are possible.

Install the device on the microscope stage. Biochip 7 is incubated with the test cell suspension for the required time. After that, the taps 4, 13 and 12 are opened and a washing liquid stream is passed, which washes off cells that have not bound immobilized antibodies in the region of test sections 30. Then, the quality of the washing is evaluated by microscopic examination of the biochip 7. (before this, the valve 12 is closed). The bound cells should remain only in the region of certain test sections 30 and be absent in the region of background sections 31. If cells that are not specifically bound to the substrate 28 from the background sections 31 are not completely eliminated, increase the flow rate of the washing liquid. To do this, increase the height of the washing liquid level, for example, by raising the capacity in which it is located, or when using a pump, increase the speed of the liquid through the device circuit.

After washing the biochip, it can be incubated with a solution that increases the binding strength of the remaining cells to the substrate. The corresponding liquid can be introduced into the container 11, for example, through the tube 18 with the open taps 12 and 13 and the closed valve 4. After the incubation is completed, the washing liquid is again passed from the container (taps 12, 13 and 4 are open).

After that, close the valve 4, open the taps 12 and 13 and let air into the circuit through the tube 18. At the same time, the liquid flows from the tank 11 and it is filled with air. Then the pressing frame 22 is pushed back and the cover 19 is removed. The removable cuvette 6 can be removed from the compartment 5. After that, if necessary, the biochip can be fixed and stained. In this case, the corresponding liquids are introduced into the container 11 (taps 12 and 13 must be closed). Then, the removable bottom 10 can be removed along with the biochip 7. The biochip 7 is separated from the bottom 10 and subjected to further research.

When working with type 2 biochips, studies of bound cells can be performed that require bringing some solutions into contact with certain areas of the biochip surface. For example, the determination of many variants of coexpression of antigens on connected cells by treating areas of the biochip surface with solutions of fluorescently-labeled antibodies. To do this, after removing the washing liquid from the tank 11 and removing the cover 19, a device 33 is installed on the surface of the biochip. In this case, channels 34 are formed that are located along the longitudinal axis of the device’s contour, but perpendicular to the test sections 30 of the second type biochip (made in the form of strips) . Each of the channels 34 crosses all test sections 30 of such a biochip. Channels 34 are filled with the necessary solutions through openings 37 using an automatic pipette, a Hamilton syringe, or other similar device. Install the lid 19 and fix it with the help of the pressing frame 22. Incubate for the required time, then wash the biochia, letting the fluid flow through the channels 34 at a low speed. To do this, open the taps 12, 13 and 4. After washing, the removable cuvette 6, a biochip 7 is removed and examined using a luminescent microscope. If necessary, the biochip 7 can be removed from the removable cell 6. When conducting other types of additional cell research, the nature and sequence of the manipulations performed may be different.

Claims (13)

1. A device for conducting analysis using a biochip, comprising a housing in which a circuit consisting of channels for supplying and discharging liquid and a compartment is placed, in which a removable cell for placing the biochip, having locking devices, and equipped with a hermetically sealed removable cover, is hermetically fixed. 2. The device according to claim 1, characterized in that it is equipped with a device locking the circuit installed in the device body. 3. The device according to claim 1, characterized in that the container for accommodating the biochip has a removable hermetically sealed bottom. 4. The device according to claims 1 and 4, characterized in that the bottom of the tank for accommodating the biochip has an adhesive coating. 5. The device according to claim 1, characterized in that it is equipped with a removable device, hermetically mounted on the surface of the biochip, and dividing the container in which the biochip is placed, into several containers of smaller volume, the bottom of each of which is formed by the surface area of the biochip. 6. The device according to claim 1, characterized in that it is equipped with a tank for washing liquid. 7. The device according to claim 1, characterized in that it is equipped with a container for collecting waste fluid. 8. The device according to claim 1, characterized in that it is equipped with a device for measuring fluid pressure. 9. The device according to claim 1, characterized in that it is equipped with a device for trapping air bubbles. 10. The device according to claim 1, characterized in that it is equipped with a device pressing the removable cover. 11. The device according to claim 1, characterized in that it is equipped with a device for additional introduction of fluid into the container in which the biochip is placed. 12. The device according to claim 1, characterized in that it is equipped with a drainage tube. 13. The device according to claim 1, characterized in that the shape and dimensions of the horizontal section of the tank for placing the biochip correspond to the shape and dimensions of the biochip.