RU2764676C1 - Apparatus with a microfluid chip for measuring the optical strength and imaging cells using the microfluid chip configuration and dynamics - Google Patents

Abstract

FIELD: testing.

SUBSTANCE: invention relates to an apparatus and method for analysing particles and imaging particles or cells in fluids, and, in particular, to an apparatus and method for imaging particles in fluids using the pressure, the dynamics of the fluid, electrokinetic and optical forces. Proposed is a microfluid chip configuration wherein injection occurs upward in the vertical direction, and the tanks with a fluid are located under the chip to minimise particle deposition in front of the analysis site and on the chip channel analysis site. According to one aspect, the fluid is input and passed upward through the lower part of the chip using a manifold, thereby eliminating orthogonal reorientation of the dynamics of the fluid. The contents of the test tubes are located under the chip and are pumped upward and vertically directly into the first channel of the chip. The long channel continues from the lower part of the chip almost to the upper part of the chip, wherein the channel is replaced by a short horizontal one. The fluid is pumped upward to the horizontal analysis site, constituting the uppermost channel/uppermost location of the fluid in the chip and therefore located near the upper part of the chip, thereby forming a clearer image.

EFFECT: creation of a method and apparatus for analysing particles and imaging particles or cells.

96 cl, 9 dwg

Description

BACKGROUND OF THE INVENTION

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

[01] In general, this invention relates to an apparatus and method for particle analysis and imaging of particles or cells in fluids, and in particular, to an apparatus and method for imaging particles in fluids using pressure, fluid dynamics, electrokinetic and optical forces.

[02] The present invention relates to a microfluidic chip in which the injection is performed in a vertical direction, and fluid tubes are located under the chip to minimize particle settling before the analysis section and in the channel analysis section of the chip.

[03] To implement the invention described in this application, changes must be made to existing microfluidic chip designs. For example, to keep the tubes upright in line with the chip, a different link must be installed compared to the link of the prior art. In particular, instead of using an inlet tube connected to the chip through an inlet at the largest front surface of the chip (as is usually done in lab-on-a-chip microfluidic systems) perpendicularly, and then pumping the fluid first transversely, then upward in The chip in the present invention has an inlet and in one aspect the fluid rises through the bottom of the chip when using the manifold, which prevents the fluid orientation/fluid dynamics from changing to horizontal.

[04] According to the invention set forth in this application, the contents of the tube is located under the chip and is pumped up and in a vertical direction directly into the first channel of the chip. The long channel extends from the bottom of the chip almost to the top of the chip. The channel is then replaced by a short horizontal one, but the new channel is so short that it is almost impossible for any cell sedimentation due to gravity and zero flow velocity on the walls. Thereafter, contrary to the prior art, the fluid is pumped up into the analysis site. Therefore, the horizontal analysis section represents the uppermost channel/highest location of the fluid in the chip and is therefore close to the top of the chip, resulting in less chip material (e.g. glass) between microscope/camera and sample than in the prior art , and therefore the formation of a clearer image. In addition, the laser suspends the cells in this channel during analysis and this prevents them from settling.

DESCRIPTION OF THE PRIOR ART

[05] According to the prior art, microfluidic chip tubes containing cells or particles to be separated and/or analyzed are placed aside and pumped horizontally into channels in the microfluidic chip. First, the contents of the tubes (eg, particles or cells) are pumped up in a vertical direction, then the direction of movement is reversed to move down, and then pumped in a horizontal direction into the chip (see, for example, US patent No. 9594071).

[06] The connection to the chip is horizontal, and this, combined with the dead volume (head space in fluid connections, which is somewhat unavoidable) in the connection, results in significant additional settling due to gravity. In addition, this configuration requires a relatively large diameter channel required for connection, and in addition to the dead volume, a region of relatively low velocity is formed, which also exacerbates the problem of particle settling. The current chip of the present invention eliminates the need for a wide horizontal inlet and a rather abrupt transition from the wide horizontal inlet to the chip's first vertical channel with relatively little upward flow. This configuration eliminates horizontal settling and the need to change direction, which causes settling. In addition, due to the entry of cells into the lower edge of the chip, the problem of settling in a dead volume is solved by orienting it in a vertical direction relative to gravity, so that cells or particles do not settle at the bottom of the horizontal channel, but instead they are constantly directed upward by the flow. This solution is not intuitive and required a lot of research to understand the problem clearly before developing a modern solution. Currently available microfluidic devices include custom or off-the-shelf connections on a polished surface and have a larger area of glass, in contrast to the present invention, which is typically exposed to any particles, such as cells contained in the sample stream, for immediate rotation after entering the chip and moving in the horizontal direction.

[07] In addition, in the prior art for cells or particles, there are several horizontal segments of the path in the microfluidic chip before reaching the analysis channel, which leads to deposition. At the moment when the contents of the tube enter the chip and the channels in the chip, the contents are pumped in a horizontal direction, and not in the vertical channel of the chip. Then the channel goes up and is replaced by a long horizontal one, and in this place the cells tend to settle to the bottom of the channel due to gravity, as well as a lower velocity near the wall due to the laminar flow regime. As such, due to the parabolic velocity profile, the flow is strongest in the middle of the channel and decreases to zero at or near the channel wall. After the first horizontal channel in the chip, the fluid turns down in front of the analysis channel where imaging and particle separation take place. Because of this configuration, there is a relatively large distance between the microscope/camera and the analysis channel. In this typical prior art configuration, the particles are forced to move downward and eventually exit the bottom of the chip.

[08] In addition, in the prior art, due to restrictive conditions, multiple horizontal paths are required, causing cells to settle at multiple locations in the channels. In turn, this causes a reduction in image quality due to the need to image through additional material at the edge of the chip. In the prior art chip, pumping up is done vertically, then horizontally, then in a zigzag line to properly suspend the cells or particles, then down and out of the chip. Zigzag channels are excluded in the present invention.

[09] In addition, the prior art discusses the reproduction of three-dimensional images of cells or particles in a fluid. For example, in M. Habaza, M. Kirschbaum, C. Guernth-Marschner, G. Dardikman, I. Barnea, R. Korenstein, C. Duschl, N. T. Shaked, Adv. Sci., 2017, 4, 1600205, shows capturing a cell, rotating it at high speed, and using interferometry to measure the refractive index distribution in the cell. In addition, interferometry is used to analyze cells in microfluidic channels (see, for example, Y. Sung et al., Phys. Rev. Appl., 2014, Feb. 27; 1:014002). However, the present invention claims to obtain multiple images of a cell or particle as it travels in fluid flow across the focal planes of the imaging device(s), thereby eliminating any need to capture the cell for 3D imaging. Other methods have been proposed such as using mechanical translation without using the bright field imaging described in this application and without using fluid flow to move the cell relative to the image plane (e.g., N. Lue et al., Opt. Express, 2008 , Sep. 29;16(20):16240-6).

[10] All prior art references cited in this application are incorporated herein by reference in their entirety.

SUMMARY OF THE INVENTION

[11] The present invention relates to a microfluidic chip that is inflated and sample vials are placed under the chip to minimize particle settling. Thus, the contents of the tube are located under the chip and are pumped upward and vertically directly into the channel of the chip. The long channel extends from the bottom of the chip to near the top of the chip. The channel then changes to a short horizontal one, but the new channel is made short enough to not exhibit any cell settling effect due to the zero flow velocity near the channel walls. In this case, contrary to the prior art, the sample is pumped upwards into the analysis section. Therefore, the horizontal section of the analysis is the uppermost channel/highest location of the fluid in the chip and is therefore close to the top of the chip, whereby less glass is between the microscope/camera and the sample than in the prior art, and therefore a clearer image. The distance of the analysis channel from the top of the chip can be from 100 µm to 2 mm, but also up to 100 mm, such as 100 µm to 200 µm, 200 µm to 300 µm, 300 µm to 400 µm, etc. In one embodiment, after the analysis section of the chip, the sample, such as fluid, cells and/or particles, is pumped down to the bottom of the chip and forced out.

[12] In addition, the present invention relates to a microfluidic chip in which horizontal path segments are minimized, especially at the point where the fluid enters the channels of the chip. Prior art chips contain horizontal channels with a total length of 13 mm (not analyzed areas), a much larger diameter injection hole at a length of about 2 mm, which enhances deposition due to low speed. According to a preferred embodiment, the chip described in this application has horizontal channels (not analyzed) in length from 0.2 to 3.0 mm, although the length of the horizontal channels can be in the range from 0.01 to 100.0 mm, such as from 0.01mm to 0.02mm, 0.02mm to 0.03mm, 0.03mm to 0.04mm, etc. The consequence of this is the channel system according to the invention, the dimensions of which are an order of magnitude different from those adopted in the prior art, which improves the flow characteristics and eliminates the deposition of cells/particles.

[13] According to another aspect, the invention relates to a microfluidic chip in which imaging and analysis occur at or near the corner of the chip, thereby improving images from multiple viewpoints due to the thinner glass and the smaller distance between the camera and the analysis channel. It also allows you to use a lens with a larger numerical aperture to improve image detail when you increase the magnification. This design reduces image distortion due to glass (or other substance that makes up the chip, such as plastic or any transparent or translucent material) and distance (eg, due to glass imperfections). The distance between the imaging device and the analysis channel can be 100 µm to 2 mm, but also up to 100 mm, such as 100 µm to 200 µm, 200 µm to 300 µm, 300 µm to 400 µm, etc.

[14] In addition, the present invention relates to a microfluidic sorting chip with separation downstream of the analysis channel, which allows simultaneous or sequential and separate use of pressure and/or laser power (or other optical power) to activate the sorting function. According to one aspect, in order to implement the sorting function, the stream from the analysis channel must be continuous. For example, particles are sent to a vertical channel and then to a horizontal sorting channel. In one embodiment, optical power and/or pressure is applied in the direction of flow in the sorting channel to move the particles along the channel. Particles that are not directly affected by the optical force are diverted into another channel due to, for example, gravity, electrokinetic forces, magnetic forces, laminar flow lines, flow lines, reduced flow velocity, orthogonal optical force, or vacuum applied to draw the particles into another channel. According to another aspect related to the sorting channel after analysis, the present invention allows the direction of the optical power from the rear side of the chip (laser or optical power is oriented in the direction of flow), and according to some aspects, the beam of this main laser is split. In embodiments, the optical power and/or pressure may be applied in a direction opposite to the direction of movement of the substance through the channel, such as upstream, or may be applied in the direction of movement of the substance in the channel, such as downstream. Sorting of cells or particles can take place in a common device or a separate chip. For example, in FIG. 1B, the fifth channel in front of the outlet tube 145 contains one or more branches, which allows one or more sorting areas.

[15] According to another aspect of the present invention, the manifold is connected to the tube such that the tube passes through the manifold and connects to the tube or other reservoir on the other side, coming into contact with the substance (eg, fluid) in the tube. The manifold allows the tubes stored under the chip to be connected to the microfluidic chip and/or the manifold allows the contents of the tubes to be pumped from the bottom of the chip while alleviating several of the problems seen in the prior art such as sedimentation of cells or particles when the tube or tube is out of the tube. connected to or in communication with the microfluidic chip.

[16] According to another aspect, the present invention relates to a microfluidic chip holder, wherein the holder comprises a structure for guiding a light source, including a built-in prism resonator, which, with the installed prism, causes light to exit at an angle relative to the chip, and this is the preferred method of illumination when limited geometry. In embodiments, the light source includes, but is not limited to, fiber optics or a collimated or focused light source. This light source is precisely directed or oriented, in particular towards the analysis channel.

[17] According to another aspect of the present invention, the device includes a second imaging device oriented orthogonally to the first camera and channel. The reasons for including a second camera are varied. In one aspect, the reason for turning on the second imaging device is to help visually align the laser or optical power in the analysis channel. According to another aspect, when using the method described in this application, data from the first camera can be recorded. With a second camera, data can be combined with data from the first camera, resulting in additional data that can be used to more accurately extrapolate the position, size, volume shape, etc. cells. This additional information about the same cell (or particle) increases the accuracy and extends the range of measurements and analysis. According to a further aspect, the second camera, in combination with the first camera, allows for a three-dimensional reconstruction of a cell or particle or group of cells or particles imaged by an orthogonal camera and a downstream or chip-side camera. When using the algorithm described in this application or otherwise, including reversing or slowing down the flow and using one or more images of a particular cell or particle or group of cells or particles, the invention allows you to analyze and process multiple images and thereby allows you to determine the characteristics/attributes/quantitative results measurements such as cell volume, cell shape, cell nucleus location, cell nucleus volume, location of organelle or intracellular inclusion, etc. According to another aspect of the present invention, the camera is oriented to obtain an image along the axis, that is, in the direction of flow.

[18] According to one aspect, the present invention does not require a tortuous or zigzag channel, which is preferred in the prior art, to keep the particles properly suspended. By vertically injecting fluid and particles or cells into the chip, the zigzag channel becomes unnecessary, vertically injected particles flow directly up the channel into the first horizontal channel (referred to in this application as the second channel).

BRIEF DESCRIPTION OF THE DRAWINGS

[19] The accompanying drawings illustrate certain aspects of some embodiments of the present invention and are not to be used to limit or define the invention. Together with the description, the drawings serve to explain some of the principles of the invention.

[20] FIG. 1A is a schematic overview of a device configuration including chip, manifold, and tubes. In FIG. 1B are schematic drawings illustrating the orientations and locations of the channels on the chip.

[21] FIG. 2A and 2B are schematic views illustrating a microfluidic chip holder according to the present invention.

[22] FIG. 3A and 3B are schematic views illustrating the positions and aspects of the chip holder according to the present invention.

[23] FIG. 4A and 4B are schematic views illustrating examples of cell pathway and configuration of imaging devices relative to the chip. In FIG. 4C is a schematic diagram illustrating another cell pathway.

[24] FIG. 5 is a schematic diagram illustrating multi-plane imaging from a chip and how it can be used to reproduce 3D images and information.

[25] FIG. 6A and 6B are schematic views illustrating multi-plane imaging of a chip in fluid flow and how it can be used to render a 3D image.

[26] FIG. 7 is a schematic diagram illustrating how a cell can be captured and/or balanced in the analysis site of a chip and displayed from multiple angles.

[27] FIG. 8 is a schematic diagram illustrating the location of the camera and light source in line with the laser and the direction of flow so that particles can move away from the camera.

[28] FIG. 9 is a schematic diagram illustrating the location of the camera and light source in line with the laser and the direction of flow so that particles can move away from the camera.

DETAILED DESCRIPTION

[29] The present invention is described with reference to specific embodiments having various features. It should be apparent to those skilled in the art that various modifications and changes may be made in the practice of the present invention without departing from the scope and spirit of the invention. One skilled in the art should be aware that these features may be used individually or in any combination to suit the requirements and conditions of a particular application or design. In addition, embodiments containing various features may consist essentially of or contain these various features. Other embodiments of the invention will become apparent to those skilled in the art upon review of the description of the invention and upon practice of the invention. The description of the invention is only exemplary or explanatory in nature and therefore changes to be made without departing from the spirit of the invention are intended to be within the scope of the invention.

[30] Before explaining in detail at least one embodiment of the invention, it should be understood that the application of the invention is not limited to the design features and arrangement of components presented in the following description or shown in the drawings. The invention is open to other embodiments or may be practiced or carried out in various ways. In addition, it should be clear that the phraseology and terminology used in this application for the purposes of description should not be construed as limiting.

[31] Referring now to the drawings, where FIG. 1A shows a general view of the device according to the idea of the invention. The sample tubes 130 are located under the microfluidic chip 100 (which in this application may also be generally referred to as the substrate) and there are no restrictions on the number or size of the tubes. The tubes are configured to hold any kind of sample that can be moved through the device, such as one or more substances, including, but not limited to, one or more of fluids, liquids, gas, plasma, serum, blood, cells, platelets, particles etc. or combinations of them. In the context of this description, the term fluid or sample can generally be used to refer to such one or more substances. The vials are connected either directly or indirectly, such as indirectly using an air conduit 110 operatively in communication with the underside of the chip. Alternatively, they may be connected using a manifold 120, also shown in FIG. 1A. In FIG. 1B shows a preferred embodiment of a microfluidic chip 100, in which a substance, fluid, particles, and/or cells are injected on one outer surface, such as the edge, of the microfluidic chip (eg, in the XZ plane of FIG. 1B). The injection is shown in Fig. 1B along one of the front surfaces separated by a small distance, such as the XZ front surface or the XY front surface. In this particular embodiment, the length of side X 160 is less than the length of side Y 170 and the length of side Z 180. This configuration ensures minimal sample deflection when entering the channels of the chip (for example, there is no need for a right turn, which is common in the current state of the art). ).

[32] FIG. 1A, a manifold 120 is shown connecting the tube or tubes 130 to the microfluidic chip 100 so that a substance, fluid, particles and/or cells can be injected or pumped into the microfluidic chip vertically in an upward direction. The manifold allows materials to be pumped from the bottom of the chip, and the electronics, flow sensors, and tubing (both liquid and air) are positioned to minimize cell settling and optimize throughput. The air line creates a pressure or vacuum which, if sealed, allows a sealed system to be created within the manifold assembly. In one embodiment, there is a distance between the tube(s) and the manifold, indicating that there is no seal and the pressure in the internal volume is atmospheric. This allows substances to be pumped out of a container or pumped into an vented container (vacuum is required for pumping from an open container). The electronics, flow sensors and tubing (both liquid and air) are located in the manifold in such a way that cell settling is minimized and thus throughput is optimized.

[33] The invention minimizes horizontal movement of the inlet tube 140 and outlet tube 145 by injecting content from the bottom of the chip, which causes problems such as particle or cell settling in the channel(s) 150. In this example, the outlet tube is offset from the inlet tube. in the Z and X directions (FIG. 1B). In embodiments, the outlet tube is offset, non-offset, straight, in line with, or inclined with respect to the inlet tube. This configuration eliminates the need for tortuous or zigzag vertical channels, as the current configuration solves prior art problems with sedimentation, such as when fluid combined with cells or particles is forced or pumped from the side into the chip in a horizontal direction. , after which the direction and dynamics of the fluid must change to move upward. In addition, the position of the manifold and the injection from the bottom of the chip allows additional elements, components, mechanisms or hardware to be located below the chip (see Fig. 1A).

[34] The manifold 120 works by controlling the air pressure above the contents in the vial and provides precise geometry to accommodate the flow sensors and electronics. A tubing, such as a fluid or air tubing, is passed through the manifold and connected to a vial on the other side. Compressed air flows through one side of the manifold and results in a closed displacement system within the manifold. By adjusting the pressure in a closed area in a system, parameters such as flow rate and fluid dynamics can be changed. The high pressure region exists in both the tube(s) 130 and the manifold 120. In another aspect, the tube does not require an air duct because it is open to the surrounding atmosphere. This allows sampling from a wider variety of containers and sources. In such an embodiment, a vacuum is applied to the other one or more tubes such that a pressure differential is created to force the fluid flow out of the open container.

[35] In one embodiment, pressure-based sample injection is used. The tubes are filled with a fluid sample and capped or with a tube connected to a chip. Prior to attaching the lid, the tube may be open to air or closed with a septum or other gas-tight element. In one aspect, the cap may contain two connections, one for a fluid, such as a gas, and one for a liquid. Optionally, an embodiment of the method also includes making a probe inlet tip connection to a sample inlet line that is in communication with the first channel.

[36] In another embodiment, vacuum-based sample injection is used. The tubes are filled with a fluid sample and capped or with a tube connected to a chip. Before attaching the lid, the tube may be open to air or closed with a septum. In one aspect, the cap may contain two connections, one for a fluid, such as a gas, and one for a liquid. Optionally, fluid may be aspirated from the tube open to atmosphere by applying a vacuum to one or more of the other tubes. Optionally, the method embodiment also includes making a probe inlet tip connection to a sample inlet line that is in communication with the first channel.

[37] FIG. 2A-B and 3A-B show a chip holder 200, 300. The chip holder includes a structure for guiding a light source 210 such as a fiber optic light source, an LED, or a laser, and a built-in prismatic resonator 220, 320 that can be installed with the prism. The light source is directed to a predetermined location or aligned with the built-in structure or channel 240 in the chip holder. The built-in space for the fiber optic light source allows illumination, such as the illumination cone 250, even in a limited geometric environment such as a microfluidic chip 230, 330, or a channel in a microfluidic chip. In particular, according to a preferred aspect, this light source is precisely directed, or oriented, or focused on the analysis channel 260. In the preferred embodiment, the chip holder includes a built-in space for the prism 220, 230 and the fiber optic light source 210 provides limited geometry illumination. In addition, the chip includes holes or openings in the base of the holder 350 for precise alignment of the fluid tube described in this application. For proper alignment, in one embodiment, adjustable screws are inserted into threaded holes 360 on one or more surfaces.

[38] FIG. 4A shows a preferred embodiment of the microfluidic chip 400 described in this application. As shown, the fluid first travels upward in the vertical direction through the first channel 410 and then enters the second horizontal channel 420. Another vertical channel 430 receives fluid even closer to the top of the chip, in which the fourth channel 440 is horizontal and, as shown in FIG. . 4 represents the analysis channel. Channels in operation are in communication with each other, which allows the sample to move through the system from one channel to another. In embodiments, the sample may flow from the first channel to the second channel, to the third channel, to the fourth channel, or vice versa, or a combination of them. A pump and/or a vacuum apparatus may be provided to apply positive and/or negative pressure to one or both of the inlet and outlet or both of the channels to allow materials to move through the channels. The analysis channel is located near one or more external surfaces of the substrate, such as front surfaces, edges or sides of the chip. For example, according to this configuration, the analysis channel is located near the top and side of the chip, thereby improving imaging and analysis of the substance in the microfluidic chip. In preferred embodiments, the analysis channel is about 1 mm to about 2 mm from the top and side of the chip. However, the analysis channel distance from the top of the chip can be 0.1mm to 100mm, such as 0.1mm to 0.2mm, 0.2mm to 0.3mm, 0.3mm to 0 .4 mm etc. In other words, the analysis channel can be located within 50%, 33%, 25%, 10% or 5% of the top of the substrate. According to the present invention, the length of the horizontal analysis channel may be from about 250 µm to about 10 mm. However, the analysis channel length may be 100 µm to 100 mm, such as 0.1 mm to 0.2 mm, 0.2 mm to 0.3 mm, 0.3 mm to 0.4 mm, etc. d. In other words, the length of the analysis channel may be about 75% or less than the height, width or length of the substrate/chip, for example, 50% or less, 33% or less, 25% or less, 10% or 5% or less of the height of the width or length of the substrate /chip.

[39] FIG. 4B shows two imaging devices 450, such as computer camcorders. In one embodiment, the camera may be located above the chip and oriented orthogonally to the analysis channel. In another embodiment, the chamber may be positioned relative to the chip side and oriented orthogonally to the flow direction or diagonally to the flow direction (eg, above the channel, below the channel, or at angles to the channel side). In embodiments, the camera may be positioned such that imaging will be performed at any angle with respect to the flow of one or more substances, such as a right angle or 90° with respect to the flow of one or more substances, or at such as from 0 to 90°, or from 10 to 80°, or 30 to 60°, etc. In another embodiment, two or more cameras may be used to image the cells or particles in the analysis channel. For example, one camera may be located above the chip and may be oriented orthogonally to the analysis channel. The second chamber may be positioned relative to the side of the chip and oriented orthogonally to the flow direction or diagonally to the flow direction (eg, above the channel, below the channel, or at an angle to the side of the channel). As shown in FIG. 4B, one or more lights 460 may be used to illuminate the analysis channel 440 and such lights may be located below the chip and shining up, relative to the side of the chip and shining in the direction of flow or opposite to the direction of flow, above the chip and shining down, or placed diagonally. in relation to the analysis channel.

[40] As shown in FIG. 8, alternatively, a dichroic mirror 840 or other suitable optical element may be used to selectively deflect light in a particular wavelength range and transmit light of other wavelengths. This will allow the cameras 810 to be in line with the analysis channel. As shown in FIG. 8 and 9, there are several embodiments that include placing the power laser 830 and the camera 810 at the same end or opposite ends of the analysis area. In addition, the camera may require a light source 860 that can be oriented in several ways, such as shown in FIG. 8 and 9. The light source may be a broad spectrum light source such as one or more LEDs, or a narrow spectrum light source such as a laser. The camera can be used as a single camera or as part of a multi-camera system in combination with other viewpoints described in this application.

[41] As shown in FIG. 4A, a light source 480, such as a laser, can be used to influence cell flow. The laser can be placed in line with the flow of cells in agreement with or against the flow of cells. In addition, the laser may be located and/or oriented orthogonally or diagonally with respect to the flow of cells.

[42] An embodiment of the invention includes a particle analysis device (see, for example, FIGS. 4-9). An embodiment of the invention includes at least one camera 450 for capturing images of particles or cells in microfluidic channels (eg, 440). In one embodiment, a laser or other power source 480, such as a collimated light source, is included that operably generates at least one beam of collimated light. At least one beam of the collimated light source has at least one beam cross section. An embodiment of the invention includes a substrate with a first channel 410 extending vertically into the substrate such that the first plane intersects the first channel 410 substantially along its length and thereby a fluid sample is injected into the substrate/chip from the bottom of the chip and forced upward by positive or negative pressure. An embodiment of the invention includes a second channel 420 orthogonal to the first channel and therefore positioned horizontally in the substrate such that the second plane intersects the second channel 420 substantially along its length and the second plane is orthogonal to the first plane. The second channel in the chip runs in the horizontal direction. The second channel is in communication with the first channel directly or indirectly. The second channel is in communication directly or indirectly with the short ascending vertical third channel 430, which refers to the network of channels close to the top of the chip. The third channel is in communication directly or indirectly with the fourth horizontal channel 440, which is located near the top and/or corner of the chip. In a preferred embodiment, the fourth channel is the channel closest to the top of the chip. In an embodiment, the fourth channel is an analysis channel. In one aspect, chamber 450 is oriented orthogonally to the direction of flow in the fourth channel. An embodiment of the invention includes a focused particle flow nozzle operatively connected to the first channel. According to another aspect of the present invention, the second channel is resized and passes through the nozzle before communicating with the third channel.

[43] FIG. 4C shows another embodiment of a probe path in a microfluidic chip. As shown, in the chip, fluid primarily flows vertically upward through the first channel 410, and this channel is directly or indirectly in communication with the second horizontal channel 420, located in this example at the top of the chip and in this embodiment representing the analysis channel . The analysis channel according to this configuration is located near the top of the chip, whereby imaging and analysis of the substance in the microfluidic chip are improved. In preferred embodiments, the analysis channel is about 1 mm to about 2 mm from the top and side of the chip. However, the analysis channel distance from the top of the chip can be 0.1mm to 100mm, such as 0.1mm to 0.2mm, 0.2mm to 0.3mm, 0.3mm to 0 .4 mm etc. In other words, the analysis channel can be located within 50%, 33%, 25%, 10% or 5% of the top of the substrate. According to the present invention, the length of the horizontal analysis channel may be from about 250 µm to about 10 mm. However, the analysis channel length may be 100 µm to 100 mm, such as 0.1 mm to 0.2 mm, 0.2 mm to 0.3 mm, 0.3 mm to 0.4 mm, etc. d. In other words, the length of the analysis channel may be about 75% or less than the height, width, or length of the substrate/chip, e.g., 50% or less, 33% or less, 25% or less, 10% or 5% or less of the height, width, or length. substrate/chip. In embodiments, the substrate may comprise one or more analysis channels, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 analysis channels.

[44] In addition, in FIG. 4C shows a light source 480, such as a laser, that can be used to influence a flow of matter, such as a flow of cells. The laser can be placed in line with the flow of cells in agreement with or against the flow of cells. In addition, the laser may be located and/or oriented orthogonally or diagonally with respect to the flow of cells.

[45] FIG. 1 and 4 the flow of fluid containing cells or particles is directed vertically through the first channel. One or more substances (fluid, cells and/or particles) enter through the bottom of the chip into the opening of the first channel and enter the substrate in a vertical direction. The first vertical channel has a length of 100 µm to 100 mm, such as 0.1 mm to 0.2 mm, 0.2 mm to 0.3 mm, and so on. The first channel is followed by a second orthogonal/horizontal channel, which in the preferred embodiment is shorter than the first channel. The second channel may have a length of 250 µm to 100 mm, such as 0.25 mm to 0.5 mm, 0.5 mm to 0.75 mm, 0.75 mm to 1.0 mm, etc. . The third channel runs vertically and parallel to the first channel. The third channel may have a length of 50 µm to 100 mm, such as 0.05 mm to 0.1 mm, 0.1 mm to 0.15 mm, 0.15 mm to 0.2 mm, etc. . Channels in operation are located in communication either directly or indirectly in such a way that the movement of one or more substances through multiple channels is ensured. The typical direction of fluid flow is shown by the flow arrows in FIG. 4A and 4C, but may be the opposite.

[46] In preferred embodiments, the fourth channel is an analysis channel, and this channel has a length of 250 µm to 100 mm, such as 0.25 mm to 0.5 mm, 0.5 mm to 0.75 mm, from 0.75 mm to 1 mm, etc. In this embodiment, the fourth channel is the closest channel to the top of the chip. The distance of the fourth channel from the top of the chip, measured vertically, can be from 100 µm to 2 mm, but also up to 100 mm, such as 100 µm to 200 µm, 200 µm to 300 µm, 300 µm to 400 µm, and etc. In other words, the fourth channel can be located within 50%, 33%, 25%, 10% or 5% of the top of the chip. An imaging device, such as a camera, may be oriented orthogonally to the fourth channel and be at a distance of about 100 µm to 2 mm from the fourth channel, but up to 100 mm, such as 100 µm to 200 µm, 200 µm to 300 µm , from 300 µm to 400 µm, etc.

[47] In one embodiment, a laser or other optical source is provided along with a focusing lens. In FIG. 4, the invention is shown with a laser 480 operating, emitting a laser beam, with the beam directed through a focusing lens into a fourth flow channel 440. The particles are aligned in the laser beam due to the presence of a gradient force that pulls the particles towards the area of the highest laser intensity. The scattering force of the laser propels the particles in the direction of propagation of the laser beam (for example, from left to right in Fig. 4A).

[48] As shown in FIG. 4B, in another embodiment, the second camera or image capture device (see 450), in addition to the camera or image capture device oriented orthogonally to the fourth channel, is oriented relative to the side of the analysis channel (or fourth) and is directed orthogonally or at any angle to the fourth channel. Obtaining one image of each cell in orthogonal views allows the calculation of numerous cell parameters, such as size and shape in two dimensions, thereby increasing the amount of information that can be captured for each cell. In addition, it allows the calculation of volumetric properties of cells, including total volume, shape, and insight into cells that are not symmetrical about an axis parallel to the direction of flow (Z-axis depicted in FIG. 5). When two or more cameras are used, a basic 3D model of cell 550 can be built by combining orthogonal images using, for example, existing 3D reconstruction algorithms such as diffraction theory or illumination rotation. The 3D model and 3D model analysis allow more accurate analysis of cell parameters such as cell size, shape, cell orientation, and other quantitative and qualitative measurements related to the particle(s) or cell(s) in the fourth channel of the microfluidic chip.

[49] FIG. 5, a portion of the analysis channel 540 is shown in two different planes, such as planes that can be displayed by an imaging device (see, for example, FIG. 4). In the first plane 520, one part of the cell or particle 510 is depicted in a specific orientation. In the second plane 530, another part of the same cell or cell is shown from another camera in a different projection. This allows multiple cell characteristics to be computed, a data matrix for each cell from each camera to be generated, and 3D imaging of the cell or particle 550 to be performed.

[50] Different portions of the cell traverse the focal planes (e.g., 630 (XZ focal plane) and 640 (YZ focal plane)) of the imaging device(s), different sections of the cell can be imaged as the cell moves, e.g., by fluid flow 620 . This is shown in FIG. 6A. In FIG. 6A, cell parts and particles are displayed at multiple points in time and/or at multiple points in space in various orientations and projections. As the cell in this example moves around the focal plane and rotates, it appears as having a different size in successive images (such as the four images for each plane from two different cameras shown exemplarily in FIG. 6A). When using 3D reconstruction algorithms, this allows for a more complex 3D image 650 of a particle or cell. From such an image, it is possible to extrapolate certain attributes of the cell or particle, such as size, volume, location of the cell and other organelles, and the size of the cell nucleus, and measure or describe the cellular topography. In addition, cell rotation can be measured as a function of power-based torque associated with changes in biophysical and biochemical properties including, but not limited to, refractive index, birefringence, cell shape and morphology.

[51] FIG. 6B shows an exemplary multiplanar imaging along with multiple images of the same cell over a period of time as the cell moves across the focal plane. In such cases, the cell image is referred to in the art as a "slice" or "image slice". An image slice is essentially the thickness of the optical image plane. The thickness of the image plane or slice is determined, among other things, by the optical magnification of the imaging system. At higher magnifications, the working distance of the lens is reduced, resulting in the need to position the lens closer to the displayed cell or particle. In one embodiment, a laser or other power source 670 may be used to influence cell movement in the assay channel. In a preferred embodiment, during imaging, cells or particles can be purposefully driven into or out of the focal plane of the channel by moving the laser and/or camera or by adjusting the flow(s) or position using hydrodynamic focusing. For example, the laser source can be moved a distance of 660 using, for example, a piezoelectric actuator or a linear electric optomechanical movable stage. This can be done for each cell or for each population. The hydrodynamic focus of the cells can be changed to affect the initial position and trajectory of the cells. For example, the particles can be aligned or oriented in the focal plane in the laser beam using a gradient force that moves the particles towards the area of highest laser intensity. The scattering force of the laser propels the particles in the direction of the laser beam propagation. See fig. 6B. Moving the laser, in this case along the x-axis, allows features to be imaged in different parts of the cell, indicated by disk 680. These may be, for example, cell nuclei, organelles, intracellular inclusions, and other cell features or particles. The laser moves the cell towards the center as a result of the action of a gradient force. In addition, it is expected that by having two or more cameras, detail and accuracy can be improved.

[52] FIG. 7, an embodiment of the invention is shown in a static mode, in which the particle or cell 710 is stopped at a certain retention location by balancing the optical force 730 and the fluid force 735. The optical force can be applied, for example, by a laser or a collimated light source. Images can be obtained in multiple planes, such as those shown in the described FIGS. 5 and 6A-B. A flow sensor is used to measure the flow rate at which each particle is stopped at a given laser power. Because the optical and fluid forces are balanced, the fluid drag force (ie, resulting from the flow rate and channel size) is equal to the optical power. In this method, the properties of each cell can be measured sequentially. Although a high performance measurement system is not available, this embodiment allows for close observation and imaging of the captured cell and also for tracking dynamic changes in refractive power resulting from biochemical and biological changes in the cell. Reagent streams containing chemicals, biological chemicals, cells, or other standard biological agents may be introduced into the flow channels to interact with the captured cell(s). These dynamic processes can be quantitatively controlled by measuring changes in refractive power during experiments on a single cell or cells.

[53] In one embodiment, the camera or other imaging device is oriented and/or focused downstream or upstream of the analysis channel such that it is in line with or parallel to the flow (see, for example, FIGS. 8 and 9). In FIG. 8 shows a camera 810 in line with the analysis channel 820 and a laser or collimated light source 830. A dichroic mirror or similar device 840 reflects the laser light 835 away from the camera to prevent damage to the camera, but lets light 865 from the illumination source 860 through to allow imaging. The chamber is oriented parallel to fluid flow 870 such that, in one embodiment, cells or particles 880 move away from the chamber. The light source is oriented orthogonally to the channel and the laser. The second dichroic mirror 845, which transmits laser light and reflects illumination light, is used to guide both illumination light and laser light through the channel. In another embodiment of this configuration, the locations of the laser and the illumination source are exchanged such that the laser is oriented orthogonally to the channel and the illumination source is parallel to the channel. The second dichroic mirror still guides both laser light and visible light through the channel.

[54] Another embodiment is shown in FIG. 9. In this case, the camera or imaging device 910 is oriented so that the cells or particles 980 move towards the camera in the fluid flow 970. Thus, the camera and laser 930 are on the same side of the channel, while the light source 960 is on the opposite side of the channel. Two dichroic mirrors 940 and 945 are used to direct laser light 935 and illumination light 965 into the channel, guide illumination light for the camera, and deflect laser light from the illumination source. In another embodiment of this configuration, the positions of the laser and camera are exchanged such that the laser is positioned orthogonal to the channel and the illumination source is positioned parallel to the channel. In such a case, the second dichroic mirror 945 directs laser light through the channel and illumination light for the camera.

[55] Optionally, an embodiment of the invention also includes at least one optical element between the source of optical power and the fourth channel, which in operation creates a standard beam mode TEM ₀₀ standard beam mode TEM ₀₁ standard beam mode TEM ₁₀ standard Hermite-Gaussian beam mode, standard Laguerre-Gaussian beam mode, Bessel beam, or standard multimode beam. Optionally, at least one optical element includes a standard cylindrical lens, a standard axicon, a standard concave mirror, a standard toroidal mirror, a standard spatial light modulator, a standard acousto-optic modulator, a standard piezoelectric mirror array, a diffractive optical element, a standard quarter-wave plate, and/or standard half wave record. Optionally, the optical power source can generate a standard circularly polarized beam, a standard linearly polarized beam, or a standard elliptically polarized beam.

[56] Optionally, a device can be implemented comprising a microfluidic channel, a source of laser light focused by optics into the microfluidic channel, and an electric field source connected to the microfluidic channel by electrodes during operation; particles in a liquid moving along a microfluidic channel; in this case, the laser light and electric field are controlled to jointly affect the particles in the microfluidic channel, as a result of which the particles are separated by size, shape, refractive index, electric charge, electric charge distribution, charge mobility, permeability and/or deformability. In yet another embodiment, the device comprises a microfluidic channel configured to apply a dielectrophoretic (DEP) field to the interior of the channel using (1) an electrode system or (2) a dielectrophoretic insulator system, and a laser light source focused by optics into the microfluidic channel; in the device, the movement of a plurality of particles in a liquid in a microfluidic channel and the combined effect of laser light and a field on particles in a microfluidic channel to capture particles or change their speed are carried out, while the dielectrophoretic field is linear or non-linear. Another possible embodiment of the device includes a microfluidic channel containing an inlet and a plurality of exits, and a source of laser light focused by optics across the microfluidic channel at a critical angle matched to the flow velocity in the microfluidic channel to generate an optical force acting on particles, while maximizing the exposure time of the laser light to the selected particles, wherein the laser light is able to perform the operation of applying forces to the particles moving through the microfluidic channel, whereby the particles are separated by a plurality of exits.

[57] Optionally, an embodiment of the invention also includes at least one particle interrogator in communication with one or more channels, such as the analysis channel(s), and in particular the fourth channel. The particle interrogator includes a standard illuminator, standard optics, and a standard sensor. Optionally, at least one particle interrogator includes a standard brightfield imager, a standard light scattering detector, a standard single wavelength fluorescence detector, a standard spectroscopic fluorescence detector, a standard CCD camera, a standard CCD camera a metal-oxide-semiconductor structure, a standard photodiode, a standard photomultiplier tube, a standard photodiode array, a standard chemiluminescent detector, a standard bioluminescent detector, and/or a standard Raman spectroscopy detector.

[58] At least one particle interrogator in communication with the fourth channel contains a laser power-based device or device that facilitates the task of identifying cell pathology, selecting and sorting cells. In one aspect, the block uses significant differences in optical pressures that result from changes in size, shape, refractive index, or morphology of the particles to separate and characterize the particles. In one aspect, a laser beam of light in the near infrared region of the spectrum applies a physical force to the cells, after which their characteristics are measured. The optical power generated by the radiation pressure, when balanced by the fluid resistance to the particles, results in particle velocity changes that can be used to identify particle differences or changes in a population of particles based on internal differences. In addition, the balance of fluid and optical forces can be used to change the position of particles relative to each other, taking into account their own properties, which allows them to be physically separated. In another embodiment, the interrogator includes a particle analysis and/or separation device, such as at least one collimated light source operatively generating at least one collimated light beam. At least one beam of collimated light includes at least one beam cross section.

[59] An embodiment of the present invention includes a combination of several of the above structural elements discussed above in a unitary device. In addition, embodiments include methods for using such a device. An example of such a unitary device is shown in Fig. 1. The embodiment shown is a 5-layer structure with 5 layers connected to each other to form a solid-state microfluidic chip, although the chip may be a single structure in contrast to the connected layers. The chip can be formed using a number of standard materials including, but not limited to, quartz glass, crown glass, borosilicate glass, soda lime glass, sapphire glass, cyclic olefin polymer (COP), polydimethylsiloxane (PDMS), OSTE, polystyrene, polymethyl methacrylate , polycarbonate, other plastics or polymers. This chip allows sample entry, hydrodynamic focusing, optical interrogation, imaging, analysis, sample exit, and full optical access of laser light into and out of areas. In addition, the chip according to the embodiments may be manufactured by 3D printing, molding, or otherwise.

[60] Optionally, at least one kind of particles includes a plurality of kinds of particles. Each particle type of the plurality of particle types includes respective intrinsic properties and respective induced properties. Optionally, intrinsic properties include size, shape, refractive index, morphology, intrinsic fluorescence, and/or aspect ratio. Optionally, the induced properties include deformation, angular orientation, rotation, rotation speed, labeled antibody fluorescence, labeled aptamer fluorescence, labeled DNA fluorescence, labeled dye fluorescence, differential retention metric, and/or gradient strength metric. This embodiment of the method also includes identifying and separating a plurality of particles according to particle types based on at least one of intrinsic properties and induced properties. Optionally, an embodiment of the method also includes interrogating or controlling the sample stream. Optionally, interrogating the sample stream includes determining at least one of the intrinsic properties and induced properties of the particle species and measuring the particle velocity of the plurality of particles. The measurement of at least one of intrinsic properties can be used for a number of applications including, but not limited to, determining the viral infectivity of a cell sample (the number of functionally infectious viral particles present in a particular cell population, similar to plaque assay or final dilution assay) for viral quantification , process development and monitoring, sample release analysis, introduced agent testing, clinical diagnostics, biomarker detection, determination of cell productivity relative to antibody or protein in process development and monitoring, determination of efficiency, quality or activation status of cells produced for cell therapy and other oncological applications , including T-cells with chimeric antigen receptors and stem cells, determination of the effect of chemicals, bacteria, viruses, antimicrobials and antivirals on a specific cell population and determination of the pathological condition or the possibility of examining it or analyzing a clinical sample of cells. Optionally, the power source has at least one beam axis and the sample flow has a sample flow axis. The step of determining at least one of the intrinsic properties and the induced properties of the particle species and the step of measuring the velocity of particles from the plurality of particles together comprise a beam axis offset from the sample flow axis. Optionally, the step of determining at least one of the intrinsic properties and the induced properties of the particle species and the step of measuring the speed of particles from a plurality of particles together comprise calculating the inclination and trajectory of a particle from a plurality of particles deviating from the axis of the sample flow to at least one axis of the beam.

[61] One skilled in the art should be aware that the disclosed features may be used individually, in any combination, or omitted based on the specifications and description of this application. When an embodiment refers to "comprising" certain features, it should be understood that an embodiment may otherwise "consist of" or "consist essentially of" any one or more features. Other embodiments will become apparent to those skilled in the art upon consideration of the description and practice of the invention.

[62] It should be especially noted that when a range of values is presented in this description, each value between the upper and lower limits of this range is also accurately disclosed. The upper and lower limits of these small ranges can be independently included or excluded. The singular forms of the indefinite and definite articles include plural objects, unless otherwise clearly indicated by the context. It is intended that the description and examples be regarded as exemplary or explanatory in nature and that variations which do not deviate from the spirit of the invention fall within the scope of the invention. In addition, all references cited in this disclosure are hereby incorporated by reference in their entirety and as such are intended to provide an efficient way to supplement the disclosure of this invention sufficient to reproduce it, as well as to provide detail on the background of the invention.

Claims (123)

1. A microfluidic chip device, comprising: a substrate containing a plurality of channels configured to transfer one or more substances, the plurality of channels comprising: the first channel located vertically in the substrate, the second channel in working communication with the first channel, located horizontally in the substrate, a third channel in working communication with the second channel located vertically in the substrate, and the fourth channel in working communication with the third channel, located horizontally in the substrate; moreover, the first, second, third and fourth channels are located in such a way that they provide a path for moving one or more substances through the substrate from the first channel to the second channel, to the third channel, to the fourth channel, and the horizontal channels are shorter in length than the first channel, and one or more substances are injected into the first channel located vertically in the substrate, by means of a flow displaced by pressure or vacuum through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel. 2. The device of claim. 1, in which the lower horizontal flat surface has at least one shorter length from one edge to the other edge compared to at least one length from one edge to the other edge of the vertical flat surface of the chip, in the vertical direction to maintain directional and volumetric continuity with the first channel. 3. The device according to claim 1, in which one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, while the lower horizontal flat surface has less than or equal surface area when compared with the surface area of the vertical flat surface of the chip, in the vertical direction to maintain directional and volumetric continuity with the first channel. 4. The apparatus of claim. 1, wherein the first channel comprises an opening located in the outer surface of the substrate and in such a way that provides a way for one or more substances to enter vertically into the substrate and to move in a vertical direction in the first channel. 5. The device according to claim 1, wherein one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel and in in which the cross section of the first channel has the same area as the cross section of the hole, has a smaller area or a larger area. 6. The apparatus of claim. 1, in which one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel and in wherein the first channel and orifice are shaped, sized, and oriented to some extent in order to maintain directional and volumetric continuity. 7. The device according to claim. 1, in addition, containing a source of collimated or focused light, oriented to interact with particles or cells in the fourth channel. 8. The apparatus of claim. 1, wherein the collimated or focused source light is oriented to propagate in a direction, against the direction, orthogonal to the direction, or diagonal to the direction of movement of one or more substances in the fourth channel. 9. The apparatus of claim. 1, wherein the fourth channel allows imaging and analysis of particles or cells in multiple focal planes. 10. The apparatus of claim. 1, wherein the fourth channel allows imaging and analysis of particles or cells in multiple focal planes during the movement of one or more substances. 11. The device of claim. 1, wherein the fourth channel allows imaging and analysis of particles or cells during the movement of one or more substances. 12. The device of claim. 1, wherein the fourth channel allows imaging and analysis of particles or cells during the movement of one or more substances and at multiple angles and/or multiple orientations. 13. The apparatus of claim. 1, wherein the fourth channel allows imaging and analysis of particles or cells during the movement of one or more substances and from multiple focal planes, at multiple angles and/or in multiple orientations by one or more imaging devices. 14. The apparatus of claim 1 further comprising one or more of an electrical force, an optical force and/or a fluid force to move the cell/cells or the particle/particles in one or more channels. 15. The device of claim 1, further comprising one or more of an electrokinetic force, an electrophoretic force, and/or a dielectrophoretic (DEP) force to move the cell/cells or particle/particles in one or more channels. 16. The apparatus of claim 1 further comprising one or more imaging devices, wherein at least one of the imaging devices is movable to some extent to change the focal plane displayed in the fourth channel. 17. The device according to claim 1, wherein the cell/cells or particle/particles in the fourth channel are able to move when the optical and/or fluid forces change, and the area of the cell displayed from the fourth channel changes as the particle moves. 18. The apparatus of claim 1, wherein the fourth channel is spaced from two or more outer surfaces of the substrate to allow imaging and analysis of multiple image slices of the cell or particle as the focal plane moves relative to the cell or particle. 19. The apparatus of claim 1, wherein the fourth channel is located at a distance from two or more external surfaces of the substrate, which allows imaging and analysis of multiple image slices of a moving cell or particle as it moves through the focal plane. 20. The apparatus of claim 1, wherein the fourth channel is located at a distance from two or more outer surfaces of the substrate, allowing imaging and analysis of multiple image slices of the suspended or static cell or particle. 21. The device according to claim 1, wherein one or more substances are capable of moving under the influence of pressure, vacuum, peristalsis, electrokinetic force, electrophoretic force, magnetic force, optical force, or any combination thereof. 22. The apparatus of claim 1, further comprising a dichroic mirror for directing light to and from the imaging device, imaging and/or analyzing cells or particles in the fourth channel. 23. The device according to claim 1, further comprising a dichroic mirror for directing collimated or focused source light to interact with cells or particles in the fourth channel. 24. The apparatus of claim. 1, further comprising a dichroic mirror for directing collimated or focused source light from an imaging device, another light source, or another part of the device. 25. The apparatus of claim 1, wherein the plurality of channels comprise a fifth channel located horizontally, vertically, or diagonally in the substrate. 26. The apparatus of claim 1, wherein the plurality of channels comprise a fifth channel that is divided into two or more channels or cavities for cell or particle sorting. 27. The device of claim. 1, wherein the fourth channel is located closer to the top of the substrate than the first channel, the second channel or the third channel. 28. The apparatus of claim. 1, wherein the fourth channel is located at a distance of 100 µm to 100 mm from the top of the substrate. 29. The device according to claim 1, wherein the first channel has a length in the range of 0.1 mm to 100.0 mm. 30. The device according to claim 1, in which the second channel has a length in the range from 0.1 mm to 100.0 mm. 31. The device according to claim. 1, in which the third channel has a length in the range from 0.05 mm to 100.0 mm. 32. The device according to claim. 1, in which the fourth channel has a length in the range from 0.1 mm to 100.0 mm. 33. The apparatus of claim 1, wherein the first channel is longer than the second channel, the third channel, or the fourth channel. 34. The device according to claim. 1, in addition, containing a cell or particle interrogation unit. 35. The device according to claim. 1, in addition, containing a channel for collecting cells or particles. 36. The apparatus of claim 1, further comprising an imaging device selected from at least one of a bright field imager, a light scattering detector, a single wavelength fluorescent detector, a spectroscopic fluorescent detector, a camera on charge-charged instruments. cameras on devices with a complementary metal-oxide-semiconductor structure, photodiode, photodiode array (PDA), spectrometer, photomultiplier or photomultiplier array, chemiluminescent detector, bioluminescent detector, standard Raman spectroscopy detection system, surface-enhanced Raman spectroscopy (SERS) detector ), a coherent anti-Stokes Raman spectroscopy (CARS) detector and/or a coherent Stokes Raman spectroscopy (CSRS) detector. 37. The device according to claim 1, in addition, containing the end of a tube that injects one or more substances into an opening located in the outer surface of the substrate and thus provides a path for entry of one or more substances into the substrate and movement in the first channel, while transverse the cross section of the hole has an area equal to, greater than or less than the cross section of the end of the tube. 38. Installation for holding a microfluidic chip, comprising: a cavity for holding the microfluidic chip; built-in light source holder to provide focused lighting in confined spaces; a built-in prism holder for allowing the prism to reflect light to a predetermined location in or on the chip; threaded screw holes to hold the microfluidic chip and allow it to be adjusted; and threaded or unthreaded passage through holes for tubing and connector or connectors. 39. The apparatus of claim 38, wherein the integrated light source is selected from at least one of a light emitting diode (LED), an organic light emitting diode (OLED), fiber optics, a fiber bundle, a laser, a flash lamp, a fluorescent lamp, and/or an incandescent lamp, or halogen lamp. 40. A device for moving cells and/or particles in a fluid, comprising: a substrate with multiple internal fluid channels, comprising: a first channel of the plurality of internal channels disposed in the substrate such that a first plane oriented vertically with respect to the substrate intersects the first channel along its length; an opening located in the outer surface of the substrate and in operative communication with the first channel so that one or more substances are able to enter the first channel and move in a vertical direction; the second channel from the plurality of internal channels, located in the substrate and in working communication with the first channel in such a way that the second plane, oriented horizontally with respect to the substrate, intersects the second channel along its length, wherein the horizontal channel is shorter in length than the first channel, and one or more substances are injected into the first channel located vertically in the substrate, by means of a flow displaced by pressure or vacuum through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel. 41. The device according to claim 40, in which one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, while the lower horizontal flat surface has at least one shorter length from one edge to the other edge compared to at least one length from one edge to the other edge of the vertical flat surface of the substrate, in the vertical direction to maintain directional and volumetric continuity with the first channel. 42. The device according to claim 40, in which one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, while the lower horizontal flat surface has less than or equal surface area when compared with the surface area of the vertical flat surface of the substrate, in the vertical direction to maintain directional and volumetric continuity with the first channel. 43. The device of claim. 40, wherein the first channel comprises an opening located in the outer surface of the substrate and in such a way that provides a way for one or more substances to enter vertically into the substrate and to move in a vertical direction in the first channel. 44. The device according to claim 40, in which one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel and in in which the cross section of the first channel has the same area as the cross section of the hole, has a smaller area or a larger area. 45. The device according to claim 40, wherein one or more substances are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, in a vertical direction to maintain directional and volumetric continuity with the first channel and in wherein the first channel and orifice are shaped, sized, and oriented to some extent in order to maintain directional and volumetric continuity. 46. The apparatus of claim 40, further comprising a source of collimated or focused light oriented to interact with particles or cells in the second channel. 47. The apparatus of claim 40, wherein the collimated or focused source light is oriented to propagate in a direction, against a direction, orthogonal to, or diagonal to the direction of travel of one or more substances in the second channel. 48. The apparatus of claim 40, wherein the second channel allows imaging and analysis of particles or cells in multiple focal planes. 49. The apparatus of claim. 40, wherein the second channel allows imaging and analysis of particles or cells in multiple focal planes during the movement of one or more substances. 50. The device of claim. 40, wherein the second channel allows imaging and analysis of particles or cells during the movement of one or more substances. 51. The apparatus of claim. 40, wherein the second channel allows imaging and analysis of particles or cells during the movement of one or more substances and at multiple angles and/or multiple orientations. 52. The apparatus of claim. 40, wherein the second channel allows imaging and analysis of particles or cells during the movement of one or more substances and from multiple focal planes, at multiple angles and/or in multiple orientations by one or more imaging devices. 53. The apparatus of claim 40, further comprising one or more of an electrical force, an optical force, and/or a fluid force to move the cell/cells or the particle/particles in one or more channels. 54. The apparatus of claim 40, further comprising one or more of an electrokinetic force, an electrophoretic force, and/or a dielectrophoretic (DEP) force to move the cell/cells or particle/particles in one or more channels. 55. The apparatus of claim 40, further comprising one or more imaging devices, wherein at least one of the imaging devices is movable to some degree to change the focal plane displayed in the second channel. 56. The apparatus of claim 40, wherein the cell/cells or particle/particles in the second channel are capable of moving as the optical and/or fluid forces change, and the area of the cell displayed from the second channel changes as the particle moves. 57. The apparatus of claim 40, wherein the second channel is spaced from two or more outer surfaces of the substrate to allow imaging and analysis of multiple image slices of the cell or particle as the focal plane moves relative to the cell or particle. 58. The apparatus of claim 40, wherein the second channel is located at a distance from two or more outer surfaces of the substrate, which allows imaging and analysis of multiple image slices of a moving cell or particle as it moves through the focal plane. 59. The apparatus of claim 40, wherein the second channel is located at a distance from two or more outer surfaces of the substrate, allowing imaging and analysis of multiple image slices of the suspended or static cell or particle. 60. The device according to claim 40, wherein one or more substances are capable of moving under the influence of pressure, vacuum, peristalsis, electrokinetic force, electrophoretic force, magnetic force, optical force, or any combination thereof. 61. The apparatus of claim 40, further comprising a dichroic mirror for directing light to and from the imaging device, imaging and/or analyzing cells or particles in the second channel. 62. The apparatus of claim 40 further comprising a dichroic mirror for directing collimated or focused source light to interact with cells or particles in the second channel. 63. The apparatus of claim 40, further comprising a dichroic mirror for directing collimated or focused source light from an imaging device, another light source, or another part of the device. 64. The apparatus of claim 40, wherein the plurality of channels comprise a third channel located horizontally, vertically, or diagonally in the substrate. 65. The apparatus of claim 40, wherein the plurality of channels comprise a third channel that is divided into two or more channels or cavities for cell or particle sorting. 66. The apparatus of claim 40 wherein the second channel is located closer to the top of the substrate than the first channel. 67. The device according to claim 40, wherein the second channel is located at a distance of 100 microns to 100 mm from the top of the substrate. 68. The device according to claim 40, wherein the first channel has a length in the range of 0.1 mm to 100.0 mm. 69. The device according to claim 40, in which the second channel has a length in the range from 0.1 mm to 100.0 mm. 70. The apparatus of claim 40, wherein the first channel is longer than the second channel. 71. The device according to claim 40, further comprising a cell or particle interrogator. 72. The device according to claim 40, further comprising a channel for collecting cells or particles. 73. The apparatus of claim 40, further comprising an imaging device selected from at least one of a bright field imager, a light scattering detector, a single wavelength fluorescent detector, a spectroscopic fluorescent detector, a camera on charge-charged instruments. cameras on devices with a complementary metal-oxide-semiconductor structure, photodiode, photodiode array (PDA), spectrometer, photomultiplier or photomultiplier array, chemiluminescent detector, bioluminescent detector, standard Raman spectroscopy detection system, surface-enhanced Raman spectroscopy (SERS) detector ), a coherent anti-Stokes Raman spectroscopy (CARS) detector and/or a coherent Stokes Raman spectroscopy (CSRS) detector. 74. The device according to claim 40, further comprising the end of a tube that injects one or more substances into an opening located in the outer surface of the substrate and thus provides a path for entry of one or more substances into the substrate and movement in the first channel, while transverse the cross section of the hole has an area equal to, greater than or less than the cross section of the end of the tube. 75. The device according to claim 1, in which one or more substances are able to move under the influence of pressure, vacuum, peristalsis, electrokinetic force, electrophoretic force, magnetic force, optical force, or any combination of them to sort the cell / cells or particles / particles into one or several separate areas or cavities. 76. The device according to claim 40, in which one or more substances are able to move under the influence of pressure, vacuum, peristalsis, electrokinetic force, electrophoretic force, magnetic force, optical force, or any combination of them to sort the cell / cells or particle / particles into one or several separate areas or cavities. 77. A method for evaluating biological particles for use in cellular immunotherapy, comprising using a device, the device comprising: using a substrate containing a plurality of channels configured to transfer one or more biological particles, the plurality of channels comprising a first channel located vertically in the substrate, the second channel in working communication with the first channel, located horizontally in the substrate, the third channel in working communication with the second channel, located vertically in the substrate, and the fourth channel in working communication with the third channel, located horizontally in the substrate, while the first, second, third and the fourth channels are arranged in such a way that they provide a path for moving one or more biological particles along the substrate from the first channel to the second channel, to the third channel, to the fourth channel; and injection of one or more biological particles into the first channel located vertically in the substrate through the lower horizontal flat surface having an opening into the first channel to maintain directional and volumetric continuity with the first channel, while the biological particles contain T cells, recombinant T cells , including CAR-T cells (chimeric antigen receptor T cells), evaluating biological particles by performing power-based measurements, moreover, power-based measurements are used to provide information regarding morphology, mobility, binding affinity, binding profiles, effect on other biological particles, effect on target cells, sensitivity to external forces such as biological, biochemical, chemical, physical and thermal influences , wherein the assessment of the biological particles comprises characterization of T cell receptors, regulatory T cells, allogeneic T cells, and bionic T cells. 78. The method of claim 77 wherein the score comprises a quantitative measurement of T cell change and provides predictive or prescriptive analytics. 79. The method of claim 77 wherein the lower horizontal flat surface has at least one shorter length from one edge to the other edge compared to at least one length from one edge to the other edge of the vertical flat surface of the substrate to maintain directional and bulk continuity with the first channel. 80. The method according to p. 77, in which one or more biological particles are injected into the first channel located vertically in the substrate, through the lower horizontal flat surface having an opening into the first channel, while the lower horizontal flat surface has less than or equal surface area at compared with the surface area of the vertical flat surface of the chip, in the vertical direction to maintain directional and volumetric continuity with the first channel. 81. The method of claim 77, wherein the first channel comprises an opening located in the outer surface of the substrate and in such a way that provides a path for one or more biological particles to enter vertically into the substrate and to move in a vertical direction in the first channel. 82. The method according to p. 77, in addition, containing a source of collimated or focused light, oriented to interact with biological particles or cells in the fourth channel. 83. The method of claim 77, wherein the collimated or focused source light is oriented to propagate in, against, orthogonal to, or diagonal to the direction of travel of one or more biological particles in the fourth channel. 84. The method according to p. 77, in which the fourth channel allows the formation of images and analysis of biological particles or cells during the movement of one or more biological particles or cells and from multiple focal planes, at multiple angles and / or in multiple orientations of one or more imaging devices. 85. The method of claim 77, further comprising one or more of electrical force, optical force, and/or fluid force to move biological particles or cells in one or more channels. 86. The method of claim 77, wherein the plurality of channels comprise a fifth channel that is divided into two or more channels or cavities for cell or particle sorting. 87. The method of claim 77, further comprising an imaging device selected from at least one of a bright field imager, a light scattering detector, a single wavelength fluorescence detector, a spectroscopic fluorescence detector, a camera on charge-charged instruments. cameras on devices with a complementary metal-oxide-semiconductor structure, photodiode, photodiode array (PDA), spectrometer, photomultiplier or photomultiplier array, chemiluminescent detector, bioluminescent detector, standard Raman spectroscopy detection system, surface-enhanced Raman spectroscopy (SERS) detector ), a coherent anti-Stokes Raman spectroscopy (CARS) detector and/or a coherent Stokes Raman spectroscopy (CSRS) detector. 88. The method of claim 77 wherein laser force cytology (LFC) is used to evaluate biological particles or cells. 89. The method of claim 88, wherein the use of laser force cytology comprises a combination of microfluidics and light-induced pressure to perform measurements comprising power, pressure, size, and velocity for each cell. 90. The method according to p. 88, in which laser force cytology is used to evaluate the characteristics of biological particles, while the characteristics contain morphology, mobility, interaction of biological particles with other physiological components, the ability to deform (to cytoskeletal changes) or the response of biological particles to environment changes. 91. The method of claim 77, wherein the evaluation of the biological particles comprises measuring a complex of one or more cells of the cell therapy preparation interacting with one or more target cells or particles. 92. A method for evaluating biological particles for use in cellular immunotherapy, comprising the use of a device, the device comprising the use of a substrate containing a plurality of channels configured to transfer one or more biological particles, the plurality of channels comprising a first channel located vertically in the substrate, the second channel in working communication with the first channel, located horizontally in the substrate, the third channel in working communication with the second channel, located vertically in the substrate, and the fourth channel in working communication with the third channel, located horizontally in the substrate, while the first, second, third and the fourth channels are arranged in such a way that they provide a path for moving one or more biological particles along the substrate from the first channel to the second channel, to the third channel, to the fourth channel; and injection of one or more biological particles into the first channel located vertically in the substrate through the lower horizontal flat surface having an opening into the first channel to maintain directional and volumetric continuity with the first channel, while the biological particles contain T cells, recombinant T cells , including CAR-T cells (chimeric antigen receptor T cells), evaluating biological particles by performing laser force cytology (LFC) based measurements, while laser force cytology based measurements are used to provide information regarding morphology, motility, binding affinity, binding profiles, effect on other biological particles, effect on target cells, sensitivity to external forces such as biological, biochemical, chemical, physical and thermal impact, wherein the assessment of the biological particles comprises characterization of T cell receptors, regulatory T cells, allogeneic T cells, and bionic T cells. 93. The method of claim 92, wherein the use of laser force cytology comprises a combination of microfluidics and light-induced pressure to produce measurements containing power, pressure, size, and velocity for each cell. 94. The method according to p. 92, in which laser force cytology is used to evaluate the characteristics of biological particles, while the characteristics contain morphology, mobility, interaction of biological particles with other physiological components, the ability to deform (to cytoskeletal changes) or the response of biological particles to environment changes. 95. The method of claim 92, wherein the evaluation of the biological particles comprises measuring a complex of one or more cells of the cell therapy preparation interacting with one or more target cells or particles. 96. The method according to p. 92, in which the evaluation of biological particles contains characterization of cells for analysis of efficiency, characterization of cells in the analysis of activity, affinity studies of CAR-T cells (T cells with a chimeric antigen receptor), detection of T cell biomarkers or marker-free detection of T-cell activation.

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