Microfluidic device, microfluidic device observation apparatus, and microfluidic device observation method

Abstract

To observe a microfluidic device by illuminating it from a side surface.SOLUTION: A microfluidic device comprises a top surface, a bottom surface, and a plurality of side surfaces. At least one of the plurality of side surfaces includes an optically transmissive portion that transmits illumination light from outside. A microfluidic device observation apparatus comprises: an illumination optical system that emits illumination light from the optically transmissive portion to the microfluidic device which has a top surface, a bottom surface, and a plurality of side surfaces, and in which at least one of the plurality of side surfaces includes the optically transmissive portion that transmits illumination light from outside; and an observation optical system that receives output light from at least one of the top surface and the bottom surface.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a microfluidic device, an observation apparatus for a microfluidic device, and a method for observing a microfluidic device. [Background technology]

[0002] Patent Document 1 describes a microfluidic device observation apparatus and a microfluidic device observation method for observing a test object present inside one or more flow channels in a microfluidic device provided with the flow channels. [Prior art document] [Patent documents] [Patent Document 1] International Publication No. 2020 / 021604 Summary of the Invention

[0003] In a first aspect of the present invention, there is provided a microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, at least one of which has an optically transparent portion that transmits illumination light from outside.

[0004] In a second aspect of the present invention, there is provided an observation apparatus for a microfluidic device, the observation apparatus comprising: an illumination optical system that illuminates the microfluidic device with illumination light from the optically transparent portion, the microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, at least one of which has an optically transparent portion that transmits illumination light from outside; and an observation optical system that receives output light from at least one of the top surface and the bottom surface.

[0005] In a third aspect of the present invention, there is provided a method for observing a microfluidic device, the method comprising: an illumination step of illuminating illumination light from the optically transparent portion of a microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, at least one of which has an optically transparent portion that transmits illumination light from outside; and an observation step of receiving output light from at least one of the top surface and the bottom surface.

[0006] The above summary of the invention does not list all of the necessary features of the present invention, and subcombinations of these features may also constitute inventions. [Brief explanation of the drawings]

[0007] [Figure 1] FIG. 1 is a perspective view showing an example of a schematic configuration of a microfluidic device 100 according to a first embodiment. [Figure 2] 1 is a side view showing an example of a schematic configuration of a microfluidic device 100 according to a first embodiment. [Figure 3] 1 shows an example of a schematic configuration of an observation device 200 for the microfluidic device 100 according to the first embodiment. [Figure 4] 10A to 10C are explanatory diagrams illustrating angle adjustment of a light sheet L1 in the microfluidic device 100 according to the first embodiment. [Figure 5] 4 is a flowchart showing the operation of the observation apparatus 200 for the microfluidic device 100 in the first embodiment. [Figure 6] FIG. 10 is a perspective view showing an example of a schematic configuration of a microfluidic device 110 according to a second embodiment. [Figure 7] FIG. 10 is a side view showing an example of a schematic configuration of a microfluidic device 110 according to a second embodiment. [Figure 8] 10 shows an example of a schematic configuration of an observation device 270 for the microfluidic device 110 according to the second embodiment. [Figure 9] 10 is a flowchart showing the operation of an observation device 270 for an observation device 110 of a microfluidic device 110 according to the second embodiment. [Figure 10] 1 shows a transmission window 131 in a first modified example. [Figure 11] 10 shows a transmission window 141 in a second modified example. [Figure 12] 10 shows a transmission window 151 in a third modified example. [Figure 13] 10 shows a transmission window 161 in a fourth modified example. [Figure 14] FIG. 10 is a diagram showing a modified example of the illumination method. [Figure 15] An example of a computer 2200 is shown. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present invention will be described below through embodiments of the invention. The following embodiments do not limit the scope of the invention. Not all of the combinations of features described in the embodiments are necessarily essential to the solution of the invention.

[0009] FIG. 1 is a perspective view showing an example of a schematic configuration of a microfluidic device 100 according to the first embodiment. Hereinafter, an XYZ coordinate system is shown in the drawings. As shown in FIG. 1, the microfluidic device 100 has a top surface 100a, a bottom surface 100b, and four side surfaces 100c to 100f. The top surface 100a faces in the direction opposite to the direction of gravity when the microfluidic device 100 is placed for observation, and the bottom surface 100b faces in the direction of gravity when the microfluidic device 100 is placed for observation. The four side surfaces 100c to 100f are surfaces that are perpendicular to the top surface 100a and the bottom surface 100b. Side 100c is the side facing the -X direction of microfluidic device 100, side 100d is the side facing the +Y direction of microfluidic device 100, side 100e is the side facing the +X direction of microfluidic device 100, and side 100f is the side facing the -Y direction of microfluidic device 100.

[0010] The microfluidic device 100 has multiple layers, and each layer of the microfluidic device 100 is provided with multiple structures 101, such as microchannels, chambers for inhabiting cells or cell aggregates, and porous membranes for co-culture. In this embodiment, the microfluidic device 100 has a substantially rectangular parallelepiped shape, but the present invention is also applicable to microfluidic devices having other three-dimensional shapes. At least one of the multiple side surfaces of the microfluidic device 100 has a transparent surface 102, which is an optically transparent portion that transmits external illumination light. The illumination light is, for example, excitation light in the visible or infrared range for fluorescence observation, or illumination light in the visible range for transmission or reflection observation. The top surface 101a or bottom surface 101b of the microfluidic device 100 has an optically transparent portion that transmits observation light. The observation light is, for example, fluorescent light in the visible range for fluorescence observation, or observation light in the visible range for transmission or reflection observation.

[0011] The microfluidic device 100 refers to a chip that places samples corresponding to biological micro-substances such as DNA, proteins, cells, cell clusters (spheroids, organoids, etc.), and tissues on a small substrate to analyze gene defects, protein distribution, reaction patterns, etc. The microfluidic device 100 in this embodiment is also called an organ on a chip, a biofunctional chip, an MPS (micro physiological systems), a biochip, a microfluidic chip, a microchip, a cell culture chip, a microchannel chip, or the like.

[0012] As an example, the microfluidic device 100 is used for culturing and analyzing cells and tissues. Furthermore, it is used to add chemical substances (drugs) and evaluate or analyze the reaction between the cultured cells. The microfluidic device 100 may include both a device in which organ cells are cultured and exhibit biological functions, and an "empty" device body in which organ cells have not yet been cultured.

[0013] The microfluidic device 100 can be fabricated using, for example, stereolithography three-dimensional printing techniques and solution cast molding processes. In addition to the above techniques, the microfluidic device 100 can also be fabricated using other microfabrication techniques, such as MEMS (Micro Electro Mechanical Systems).

[0014] Each layer of the microfluidic device 100 is formed, for example, by a substrate. The substrate may be formed, for example, of glass. The substrate may be formed, for example, of a resin material such as polymethyl methacrylate (PMMA), polycarbonate (PC), cycloolefin copolymer (COC), cycloolefin polymer (COP), polystyrene (PS), or silicone. The microfluidic device 100 may have a cover that covers the entire microfluidic device 100.

[0015] FIG. 2 is a side view showing an example of a schematic configuration of the microfluidic device 100 according to the first embodiment. FIG. 2 is a view of the microfluidic device 100 of FIG. 1 as viewed from the −X direction, showing a side surface 100c of the microfluidic device 100. In the microfluidic device 100 according to the first embodiment, the entire side surface 100c serves as a transmissive surface 102 that transmits external illumination light. The external illumination light is light that illuminates the microfluidic device 100 when observing the microfluidic device 100 with an observation device such as a microscope. Therefore, the transmissive surface 102 transmits external illumination light. The transmissive surface 102 is made of a material that transmits external illumination light, such as glass.

[0016] FIG. 3 shows an example of the schematic configuration of an observation device 200 for a microfluidic device 100 in the first embodiment. The observation device 200 in this embodiment is an inverted microscope, as an example. However, an upright microscope is also applicable. The observation device 200 illuminates the microfluidic device 100 with a sheet of light L1, which is excitation light, and captures the fluorescence excited from the fluorescent dye in the observation device 200 to obtain an image of the observation device 200. The sheet of light L1 is illumination light having a sheet-shaped illumination area.

[0017] 3, the observation device 200 has a sheet illumination optical system 210, an observation optical system 220, a stage 230, and a PC (control device) 240. The microfluidic device 100 is placed on the stage 230 of the observation device 200, and the microfluidic device 100 can be moved in the up-down and left-right directions (XYZ directions) by moving the stage 230 in the up-down and left-right directions (XYZ directions). The sheet illumination optical system 210, the observation optical system 220, and the stage 230 are connected to the PC 240 and can be automatically controlled.

[0018] The sheet illumination optical system 210 includes an optical fiber 211 that propagates laser light emitted from a laser light source (not shown), a collector lens 212, a cylindrical lens 213, and a variable aperture 214. The sheet illumination optical system 210 is an illumination optical system that illuminates cells and the like inside the microfluidic device 100. The sheet illumination optical system 210 is disposed on the −X direction side of the side of the microfluidic device 100 and emits a light sheet L1 toward the microfluidic device 100. The exit end surface of the optical fiber 211, a component of the sheet illumination optical system 210, constitutes a point light source and is irradiated with laser light. The laser light is shaped into a sheet-shaped light sheet L1 by the cylindrical lens 213, and the sheet light is limited by the variable aperture 214 and illuminated toward the observation region 300 of the microfluidic device 100. The longitudinal direction of the light sheet L1 is perpendicular to the optical axis of the sheet illumination optical system 210. The observation area 300 is the area of ​​the microfluidic device 100 that a user wishes to observe under a microscope.

[0019] The light sheet L1 enters the microfluidic device 100 from the side surface 100c of the microfluidic device 100 and illuminates cells and the like inside the microfluidic device 100. In FIG. 3, the light sheet L1 illuminates the microfluidic device 100 parallel to the X-axis. As shown in FIG. 3, the output end of the optical fiber 211 is configured to be movable (shiftable) in the height direction (±Z direction) from the optical axis of the sheet illumination optical system 210. This allows the illumination area of ​​the light sheet L1 to be configured to be movable in the height direction (±Z direction). It is desirable to automatically fine-tune the intensity and longitudinal width of the light sheet L1 so as to minimize ghosts caused by diffuse reflection from the structure 101 inside the microfluidic device 100.

[0020] The observation optical system 220 has an objective lens 221, a second objective lens (imaging lens) not shown, and a two-dimensional detector 222. The objective lens 221 may have a configuration that allows for electric switching between low magnification and medium-high magnification. The light sheet L1 emitted from the sheet illumination optical system 210 illuminates cells in the observation region 300 of the microfluidic device 100. The observation region 300 is not limited to cells, and may also include cell clusters and tissues. The cells in the observation region 300 are excited by the light sheet L1 and emit fluorescence L2, which is output light. The fluorescence L2 is emitted from the bottom surface 100b of the microfluidic device 100. The bottom surface 100b of the microfluidic device 100 is made of a material that transmits the fluorescence L2. The microfluidic device 100 may be observed from the upper surface 100a direction. In this case, the observation optical system 220 is disposed in the direction of the upper surface 100a of the microfluidic device 100, and the upper surface 100a is made of a material that transmits the fluorescence L2.

[0021] The fluorescence L2 emitted from the bottom surface 100b of the microfluidic device 100 is collected by an objective lens 221 and detected by a two-dimensional detector 222. The two-dimensional detector 222 is, for example, an image sensor such as a CCD (Charge Coupled Device) image sensor or an sCMOS (Scientific Complementary Metal Oxide Semiconductor) image sensor. The observation optical system 220 may further include other optical members such as a condenser lens or a dichroic mirror.

[0022] The PC (control device) 240 includes a CPU and a memory (storage unit), and the CPU reads and executes a control program stored in the memory, thereby controlling the operation of the observation device 200. As indicated by the dashed-dotted line in Fig. 3, the PC 240 is connected to the stage 230 of the microscope and is capable of controlling the operation of the entire microscope. The PC 240 has an input unit that receives various instructions and settings from the user and transmits them to the control unit of the PC 240, and a display unit that receives commands from the control unit of the PC 240 and displays a GUI (Graphical User Interface) screen, various dialog boxes, and the like to the user.

[0023] FIG. 4 is an explanatory diagram of angle adjustment of the light sheet L1 of the microfluidic device 100 in the first embodiment. FIG. 4 is a partially enlarged view of the observation device 200, illustrating only some of the components of the observation device 200. In FIG. 3, the light sheet L1 incident on the microfluidic device 100 from the sheet illumination optical system 210 is parallel to the X-axis. However, as shown in FIG. 4, the angle of the light sheet L1 can be adjusted by adjusting the angle (direction) (tilt) between the exit end face of the optical fiber 211 and the optical axis of the sheet illumination optical system 210. In the example shown in FIG. 4, the exit end of the optical fiber 211 faces obliquely upward, and the light sheet L1 illuminates the observation area 300 at an obliquely downward angle via the collector lens 212 and the cylindrical lens 213. As shown in FIG. 4, by adjusting the angle of the light sheet L1, for example, the observation area 300 can be illuminated from an appropriate angle. Furthermore, when an obstacle such as a structure 101 is present between the observation area 300 and the sheet illumination optical system 210, the obstacle can be avoided to illuminate the observation area 300. Adjustment of the direction (tilt) of the emission end of the optical fiber 211 and adjustment of the height position (shift) (position in the Z direction) of the emission end of the optical fiber 211 can be performed in combination.

[0024] 5 is a flowchart showing the operation of the observation apparatus 200 for the microfluidic device 100 in the first embodiment. In step S01, the user places the microfluidic device 100 on the microscope stage 230. Then, in step S02, the user selects the type of the microfluidic device 100 on the GUI.

[0025] Next, in step S03, the PC 240 determines whether it has information about the type of microfluidic device 100 selected by the user. The information about the microfluidic device 100 may include, for example, at least one of the following: the shape, structure, size, manufacturer, cultured cell information, and model number of the microfluidic device 100. If the PC 240 has the information about the microfluidic device 100 (step S03: YES), the process proceeds to step S05, where the information about the microfluidic device 100 is read, and the user selects an assay compatible with the microfluidic device 100 from among the multiple assays displayed on the GUI. If the PC 240 does not have the information about the microfluidic device 100 (step S03: NO), the process proceeds to step S04, where an error message indicating that the microfluidic device 100 is an incompatible microfluidic device is displayed on the GUI. An assay is an image acquisition and analysis routine that acquires an image and performs predetermined analysis and evaluation processing on the acquired cell image to obtain a predetermined result, such as counting the number of cells, calculating cell density distribution, and determining whether the cells are viable or dead.

[0026] Next, in step S06, when the user presses the GO button on the GUI, the process proceeds to step S07, where the observation device 200 moves the stage 230 to move the field of view of the objective lens 221 to the first observation region 300. The position of the observation region 300 can be specified by the user with respect to the three-dimensional image of the entire microfluidic device 100 displayed on the GUI, and the processing of step S07 is automatically performed by the observation device 200 based on the specified position. The processing of step S07 may also be performed manually by the user.

[0027] Next, in step S08, the observation conditions (including the illumination conditions, the magnification of the objective lens 221, the relative position between the objective lens 221 and the observation region 300, etc.) are switched to acquire an image of the observation region 300, and the image of the observation region 300 is acquired. When acquiring the image, the user sets optimal illumination conditions for the transmission window corresponding to the first observation region 300 while taking the structure 101 into consideration by referring to the 3D image of the entire microfluidic device 100 displayed on the GUI, and irradiates the transmission window with the light sheet. Note that by linking information about the observation region 300, information about the structure 101, and information about the transmission window and storing them in the PC 240, the user may be able to irradiate the transmission window corresponding to the first observation region 300 with the light sheet under optimal illumination conditions simply by specifying the observation region 300. The illumination conditions include, for example, the intensity of the light sheet, the longitudinal width of the light sheet, the illumination position, the illumination angle, the illumination wavelength, and the size of the variable aperture corresponding to the longitudinal width of the light sheet. In step S09, it is confirmed whether the image quality of the acquired image of the observation area 300 is good, and if it is good (step S09: YES), the process proceeds to the next step S10. If the image quality is not good (step S09: NO), the process returns to step S08 and the observation conditions are reset.

[0028] Next, in step S10, it is determined whether the next observation region exists. If the next observation region exists (step S10: YES), the process returns to step S07 and repeats the processes from step S07 to S09. If the next observation region does not exist (step S10: NO), the process proceeds to the next steps S11 and S12.

[0029] In step S11, assay image processing (image analysis) is performed, and in step S12, the processing results (analysis results) are saved.

[0030] According to the microfluidic device 100 of the first embodiment, the side surface 100c of the microfluidic device 100 has a transmitting surface 102 that is an optically transmitting portion that transmits illumination light from outside. This allows a light sheet L1 for observing the microfluidic device 100 to enter the inside of the microfluidic device 100 from the side surface 100c of the microfluidic device 100. Therefore, even if, for example, a structure 101 such as a microchannel is installed on the top surface 101a of the microfluidic device 100 and it is difficult to observe the desired observation region 300 from above, the light sheet L1 can be made to enter from the side surface 100c to perform observation.

[0031] Similarly, even if the internal structures 101 of the microfluidic device 100 are stacked in the vertical direction, making it difficult to perform pinpoint transmission observation of the desired observation area 300 from above, or if the microfluidic device 100 is composed of multiple layers and is thick in the vertical direction, making it difficult to perform transmission observation of the desired observation area 300 from above, it is possible to perform observation by irradiating sheet light L1 from the side surface 100c.

[0032] According to the microfluidic device 100 of the first embodiment, it is possible to adjust the angle of incidence of the light sheet L1, which is used to observe the microfluidic device 100, with respect to the transmission window. This allows the observation region 300 to be observed with good contrast, avoiding the structures 101, such as microchannels, in the microfluidic device 100.

[0033] According to the observation apparatus 200 for the microfluidic device 100 of the first embodiment, it is possible to achieve the same effects as the microfluidic device 100 of the first embodiment.

[0034] Fig. 6 is a perspective view showing an example of a schematic configuration of a microfluidic device 110 according to the second embodiment. In the following description of the microfluidic device 110 according to the second embodiment, the same components as those in the microfluidic device 100 according to the first embodiment are denoted by the same reference numerals, and description thereof will be omitted. As shown in Fig. 6, the microfluidic device 110 according to the second embodiment has a side surface 100c formed with transmission windows 121 and 123, which are optically transmissive portions that transmit illumination light from outside.

[0035] FIG. 7 is a side view showing an example of a schematic configuration of a microfluidic device 110 according to the second embodiment. FIG. 7 is a view of the microfluidic device 110 of FIG. 6 as viewed from the −X direction, illustrating a side surface 100c of the microfluidic device 110. A transmission window 121 and a transmission window 123 are formed on the side surface 100c of the microfluidic device 100 according to the second embodiment. The transmission window 121 is provided in the lower left portion of the side surface 100c, and the transmission window 123 is provided in the upper right portion of the side surface 100c. The transmission windows 121 and 123 are provided corresponding to the positions of the regions of the microfluidic device 110 to be observed. The transmission windows 121 and 123 are made of a material that transmits external illumination light for observing the microfluidic device 110, such as glass. The transmission windows 121 and 123 according to the second embodiment have a flat shape with a constant thickness in the X direction.

[0036] Because the size of the transmission window 121 and the transmission window 123 is sufficiently small compared to the entire area of ​​the side surface 100c, it takes time to locate the position of the transmission window 121 or the transmission window 123, and it is also difficult to ensure the accuracy of detection. For this reason, positioning markers 122 and 124 are formed near the transmission window 121 and the transmission window 123 (for example, near the four corners). The vicinity of the transmission window refers to the outside or inside of the transmission window. This is not limited to the vicinity of the four corners, but may also be two corners on a diagonal line. The positioning markers 122 and 124 are used to locate and identify the positions of the transmission window 121 and the transmission window 123. The positioning markers 122 and 124 may be formed of a material (metal deposition film, dielectric multilayer film) that has the property of reflecting IR (infrared) light irradiated from an IR illumination light source 250 (described later), or may be formed of a material that scatters IR light, or may be ground glass that scatters IR light. The positioning marker 122 and the positioning marker 124 differ in shape, color, or material.

[0037] 8 shows an example of a schematic configuration of an observation device 270 for the microfluidic device 110 according to the second embodiment. As shown in FIG. 8, the observation device 270 includes a sheet illumination optical system 210, an observation optical system 220, a stage 230, a PC 240, an IR illumination light source 250, and a marker observation optical system 260. Compared to the microfluidic device 100 according to the first embodiment, the microfluidic device 110 according to the second embodiment further includes the IR illumination light source 250 and the marker observation optical system 260. The IR illumination light source 250 and the marker observation optical system 260 are used to check the illumination state of the sheet light L1 from the sheet illumination optical system 210.

[0038] The IR illumination light source 250 emits IR light toward at least one of the transmission window 121, in which the positioning marker 122 and the positioning marker 124 are provided, and the transmission window 123. The IR illumination light source 250 includes, for example, at least one infrared light emitting diode (LED). The IR illumination light source 250 may include multiple LEDs. The IR light is reflected or partially absorbed by at least one of the positioning marker 122 on the transmission window 121 and the positioning marker 124 on the transmission window 123 illuminated with the IR light, and then enters the marker observation optical system 260.

[0039] The marker observation optical system 260 has an IR detector 261, a lens 262, and a dichroic mirror 263. The dichroic mirror 263 has the property of transmitting the light sheet L1 and reflecting IR light. The range that the marker observation optical system 260 can detect all at once is the range that encompasses the largest transmission window 121 and its positioning marker 122. The IR detector 261 is a detector that detects IR light, and is, for example, an image sensor such as an IR-CCD (Charge Coupled Device) image sensor or an IR-CMOS (Complementary Metal Oxide Semiconductor) image sensor. The IR light that enters the marker observation optical system 260 is reflected by the dichroic mirror 263, collected by the lens 262, and enters the IR detector 261. Each of the positioning markers 122 and 124 is made up of a plurality of members, but these members can be illuminated and observed all at once, so the positions of the transmission windows 121 and 123 can be quickly identified.

[0040] The position of the transmission window 121 can be identified by detecting the image of the positioning marker 122 with the IR detector 261. Therefore, for example, if the light sheet L1 travels on the optical axis of the sheet illumination optical system 210 and is incident on the -Z direction side of the position of the transmission window 121, and the desired observation region 300 cannot be illuminated, the light sheet L1 can be made to illuminate the desired observation region 300 through the transmission window 121 by moving the stage 230 in the -Z direction or by moving the height position of the end face of the optical fiber 211 of the sheet illumination optical system 210 in the +Z direction.

[0041] 9 is a flowchart showing the operation of the observation apparatus 270 of the microfluidic device 110 in the second embodiment. The operations of steps S01 to S12 of the observation apparatus 270 in the second embodiment are the same as the operations of steps S01 to S12 of the observation apparatus 200 in the first embodiment. The observation apparatus 270 in the second embodiment further includes operations of steps S13 and S14.

[0042] In step S13, a positioning marker (e.g., positioning marker 122) of a transmission window (e.g., transmission window 121) is detected by the marker observation optical system 260. By detecting the positioning marker 122, the position of the transmission window 121 can be identified. In the next step S14, optimal illumination conditions are set based on information about the structure 101 associated with the identified transmission window 121 and information about the observation area 300. The illumination conditions include, for example, the intensity of the light sheet, the width of the light sheet in the longitudinal direction, the illumination position, the illumination angle, the illumination wavelength, and the size of the variable aperture corresponding to the width of the light sheet in the longitudinal direction.

[0043] At least one of the transmission windows 121 and 123 may have an optical function of collecting the sheet light L1 or controlling the traveling direction of the sheet light L1. Modifications having such optical functions will be described below.

[0044] FIG. 10 shows a transmission window 131 in a first modified example. FIG. 10 illustrates only a side surface 100c of the microfluidic device 110, and other components are omitted. FIG. 10 shows a cross-sectional view of the side surface 100c. A convex portion 132 having a convex lens function is integrally formed with the transmission window 131 in the first modified example, and thus the transmission window 131 has a convex lens function as an example of an optical function. The convex lens function of the transmission window 131 makes it possible to collimate light from an optical fiber or the like, for example. Positioning markers 133 are formed at the four corners of the transmission window 131.

[0045] FIG. 11 shows a transmissive window 141 in a second modified example. In FIG. 11, only the side surface 100c of the microfluidic device 110 is illustrated, and other components are omitted. FIG. 11 shows a cross-sectional view of the side surface 100c. The transmissive window 141 in the second modified example has a recessed portion 142 integrally formed therewith, which functions as a concave lens. This allows the transmissive window 141 to have a concave lens function, which is an example of an optical function. An optical scanner 144 is disposed on the −X direction side of the transmissive window 141, scanning the light sheet L1 incident on the transmissive window 141. The optical scanner 144 can deflect the light sheet L1 in multiple directions by scanning the light sheet L1. Because the transmissive window 141 has a concave lens function, for example, when the light sheet L1 is scanned by the optical scanner 144, the light beam is not refracted at the interface. This allows the incident angle of the light sheet L1 to be changed as scanned, and also provides the effect of maintaining a constant thickness of the light sheet L1. In addition, when it is desired to illuminate with a wide illumination light, the light flux of the sheet light L1 can be widened. Positioning markers 143 are formed at the four corners of the transmission window 141.

[0046] FIG. 12 shows a transmissive window 151 in a third modified example. In FIG. 12, only the side surface 100c of the microfluidic device 110 is illustrated, and other components are omitted. FIG. 12 shows a cross-sectional view of the side surface 100c. The transmissive window 151 in the third modified example has a recessed portion 152 integrally formed therewith. The recessed portion 152 includes an inclined flat surface 152a inclined relative to the side surface and a curved surface 152b. The inclined flat surface 152a has, as an example of an optical function, a function of allowing illumination light from a specific direction to enter without refracting it. The curved surface 152b has, as an example of an optical function, a concave lens function. An optical scanner 154 is disposed on the −X direction side of the transmissive window 151. The optical scanner 154 scans the light sheet L1 entering the transmissive window 151, and can change the angle of incidence by scanning the light sheet L1. The curved surface 152b of the transmissive window 151 has a concave lens function, allowing the transmissive window 151 to have the same function as the transmissive window 141 in the second modified example. Furthermore, a plurality of observation regions 300 can be collectively illuminated at a desired angle through the transmission window 151. Positioning markers 153 are formed at the four corners of the transmission window 151.

[0047] FIG. 13 shows a transmission window 161 in a fourth modified example. In FIG. 13, only the side surface 100c of the microfluidic device 110 is illustrated, and other components are omitted. FIG. 13 shows a cross-sectional view of the side surface 100c. The transmission window 161 in the fourth modified example has a plurality of masks 162 formed thereon, which block part of the light sheet L1 (in a direction in which irradiation is not desired). This gives the transmission window 161 a light-shielding function, which is an example of an optical function. The light-shielding function of the transmission window 161 makes it possible, for example, to adjust the size of the beam of the light sheet L1. Furthermore, by collectively illuminating the microfluidic device 110 and generating multiple beams in the irradiation direction using the masks 162, multiple observation regions 300 can be illuminated simultaneously. Positioning markers 163 are formed at the four corners of the transmission window 161.

[0048] 10 to 13 may be combined in any manner. For example, a mask 162 may be combined with a transmission window 141 having a concave lens function. Alternatively, a mask 162 may be combined with a transmission window 151 having a plurality of inclined flat surfaces 152a and curved surfaces 152b.

[0049] The microfluidic device 110 of the second embodiment can achieve the same effects as the microfluidic device 100 of the first embodiment.

[0050] According to the microfluidic device 110 of the second embodiment, a plurality of transmission windows 121 and 123 are formed on the side surface 101c, which makes it easy to observe a plurality of observation regions 300 at different positions.

[0051] According to the microfluidic device 110 of the second embodiment, the entire side surface 101c is not a transparent surface, but a plurality of transparent windows 121 and 123 are formed in a part of the side surface 101c. This allows the other part of the side surface 101c to be made of any material, thereby reducing material costs.

[0052] According to the microfluidic device 110 of the second embodiment, positioning markers 122 and 124 for identifying the positions of the transmission windows 121 and 123 are formed at the four corners of the transmission windows 121 and 123, and the observation device 270 further includes an IR illumination light source 250 and a marker observation optical system 260 for detecting the positioning markers 122 and 124. This allows the transmission windows 121 and 123 to be detected quickly.

[0053] According to the microfluidic device 110 of the second embodiment, the transmission windows 131 to 161 of the first to fourth modifications have various optical functions, which make it possible to optically manipulate the sheet light L1 from the sheet illumination optical system 210. Note that the transmission surface 102 of the first embodiment may also be provided with an optical function.

[0054] According to the observation device 270 for the microfluidic device 110 in the second embodiment, it is possible to achieve the same effects as the microfluidic device 110 in the second embodiment.

[0055] In the first and second embodiments described above, an optically transparent portion is provided on the side surface 101c of the microfluidic device 100, 110, and the observation region 300 is illuminated from the side surface 101c. However, a reflecting mirror may be provided inside the microfluidic device 100, 110 to change the direction of the sheet light L1 illuminated from the side surface 101c. For example, the sheet light L1 illuminated from the side surface 101c is light that travels in a direction generally parallel to the XY plane, but the sheet light L1 illuminated from the side surface 101c may be reflected by a reflecting mirror and directed in the Z direction.

[0056] In the first and second embodiments, an optically transparent portion is provided on the side surface 101c of the microfluidic device 100, 110, and the inside is observed by irradiating the microfluidic device 100, 110 with the light sheet L1 from the side surface 101c. However, the light sheet L1 may also be irradiated from the top surface 101a or the bottom surface 101b of the microfluidic device 100, 110, or a combination of irradiating the microfluidic device 100, 110 with the side surface 101c and irradiating the microfluidic device 100, 110 with the top surface 101a or the bottom surface 101b may be implemented.

[0057] In the first and second embodiments, the transmittance of illumination light on the side surfaces other than the side surface 101c on which the transmission surface and the transmission window are formed may be intentionally made lower than that of the side surface 101c on which the transmission surface and the transmission window are formed. For example, the side surfaces may be roughened to make them easier to grip with a human hand. Furthermore, the side surfaces may be covered with a light-shielding mask to prevent external light from the side surfaces from affecting the cells inside.

[0058] In the second embodiment, two transmission windows, 121 and 123, are formed on one side surface 100c of the microfluidic device 110. However, the number of transmission windows formed on one side surface may be three or more. Also, multiple transmission windows may be formed on multiple side surfaces. In this case, it is desirable to provide the sheet illumination optical system 210 corresponding to the multiple side surfaces on which the transmission windows are formed.

[0059] In the second embodiment, a transmission window is formed on the side surface 100c of the microfluidic device 110, and positioning markers are formed at the four corners of the transmission window. However, the positioning markers may be formed inside the microfluidic device 110 instead of near the transmission window. Furthermore, the markers may be formed at a location other than the four corners of the transmission window. The number of markers formed does not have to be four.

[0060] FIG. 14 is a diagram showing a modified illumination method. In the first and second embodiments, a sheet illumination optical system 210 is provided on the side surface of the microfluidic device 100, 110, and a light sheet L1 is irradiated from the side surface of the microfluidic device 100, 110 to observe the interior. However, as shown in FIG. 14, instead of the sheet illumination optical system 210, an optical fiber 301 that emits light as a whole or a light source unit (not shown) having multiple LEDs arranged in a line may be arranged parallel to the side surface of the microfluidic device 100, 110 (in the XY plane) to illuminate the interior. In this case, it is desirable to provide a transmission surface or a transmission window corresponding to the side surface on which the optical fiber 301 is arranged. The optical fiber 301 and the light source unit may be configured to be movable in the Z direction depending on the position of the transmission window and the position of the observation area. Dark-field illumination and bright-field illumination can be achieved by adjusting the Z position.

[0061] Various embodiments of the present invention may also be described with reference to flowcharts and block diagrams, where the blocks may represent (1) stages of a process in which operations are performed or (2) sections of an apparatus responsible for performing the operations. Particular stages and sections may be implemented by dedicated circuitry, programmable circuitry provided with computer-readable instructions stored on a computer-readable medium, and / or a processor provided with computer-readable instructions stored on a computer-readable medium. Dedicated circuitry may include digital and / or analog hardware circuitry, and may include integrated circuits (ICs) and / or discrete circuits. Programmable circuitry may include reconfigurable hardware circuitry, including logical AND, OR, XOR, NAND, NOR, and other logic operations, flip-flops, registers, memory elements such as field programmable gate arrays (FPGAs), programmable logic arrays (PLAs), and the like.

[0062] A computer-readable medium may include any tangible device capable of storing instructions that are executed by an appropriate device, such that the computer-readable medium having instructions stored thereon comprises an article of manufacture containing instructions that can be executed to create means for performing the operations specified in the flowcharts or block diagrams. Examples of computer-readable media may include electronic, magnetic, optical, electromagnetic, and semiconductor storage media. More specific examples of computer-readable media may include floppy disks, diskettes, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), electrically erasable programmable read-only memory (EEPROM), static random access memory (SRAM), compact disc read-only memory (CD-ROM), digital versatile disc (DVD), Blu-ray (RTM) disc, memory stick, integrated circuit card, and the like.

[0063] The computer readable instructions may include either assembler instructions, Instruction Set Architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state-setting data, or source or object code written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Smalltalk®, JAVA®, C++, etc., and conventional procedural programming languages ​​such as the “C” programming language or similar programming languages.

[0064] The computer-readable instructions may be provided to a processor or programmable circuitry of a general-purpose computer, special-purpose computer, or other programmable data processing apparatus, either locally or over a wide-area network (WAN) such as a local area network (LAN), the Internet, etc., which executes the computer-readable instructions to create means for performing the operations specified in the flowcharts or block diagrams. Examples of processors include computer processors, processing units, microprocessors, digital signal processors, controllers, microcontrollers, etc.

[0065] 15 illustrates an example of a computer 2200 in which aspects of the present invention may be embodied, in whole or in part. Programs installed on the computer 2200 may cause the computer 2200 to function as or perform operations associated with an apparatus or one or more sections of the apparatus according to embodiments of the present invention, and / or to perform a process or steps of a process according to embodiments of the present invention. Such programs may be executed by the CPU 2212 to cause the computer 2200 to perform specific operations associated with some or all of the blocks in the flowcharts and block diagrams described herein.

[0066] A computer 2200 according to this embodiment includes a CPU 2212, a RAM 2214, a graphics controller 2216, and a display device 2218, which are interconnected by a host controller 2210. The computer 2200 also includes input / output units such as a communication interface 2222, a hard disk drive 2224, a DVD-ROM drive 2226, and an IC card drive, which are connected to the host controller 2210 via an input / output controller 2220. The computer also includes legacy input / output units such as a ROM 2230 and a keyboard 2242, which are connected to the input / output controller 2220 via an input / output chip 2240.

[0067] The CPU 2212 operates according to programs stored in the ROM 2230 and RAM 2214, thereby controlling each unit. The graphics controller 2216 acquires image data generated by the CPU 2212 into a frame buffer or the like provided in the RAM 2214 or into the graphics controller 2216 itself, and causes the image data to be displayed on the display device 2218.

[0068] The communication interface 2222 communicates with other electronic devices via a network. The hard disk drive 2224 stores programs and data used by the CPU 2212 in the computer 2200. The DVD-ROM drive 2226 reads programs or data from the DVD-ROM 2201 and provides the programs or data to the hard disk drive 2224 via the RAM 2214. The IC card drive reads programs and data from an IC card and / or writes programs and data to an IC card.

[0069] The ROM 2230 stores therein a boot program or the like that is executed by the computer 2200 upon activation, and / or programs that depend on the hardware of the computer 2200. The input / output chip 2240 may also connect various input / output units to the input / output controller 2220 via a parallel port, a serial port, a keyboard port, a mouse port, etc.

[0070] The programs are provided by a computer-readable medium such as a DVD-ROM 2201 or an IC card. The programs are read from the computer-readable medium, installed in the hard disk drive 2224, RAM 2214, or ROM 2230, which are also examples of computer-readable media, and executed by the CPU 2212. Information processing described in these programs is read by the computer 2200, and brings about cooperation between the programs and the various types of hardware resources described above. An apparatus or method may be configured by realizing information manipulation or processing in accordance with the use of the computer 2200.

[0071] For example, when communication is performed between the computer 2200 and an external device, the CPU 2212 may execute a communication program loaded into the RAM 2214 and instruct the communication interface 2222 to perform communication processing based on the processing described in the communication program. Under the control of the CPU 2212, the communication interface 2222 reads transmission data stored in a transmission buffer processing area provided in the RAM 2214, the hard disk drive 2224, the DVD-ROM 2201, or a recording medium such as an IC card, and transmits the read transmission data to the network, or writes reception data received from the network to a reception buffer processing area or the like provided on the recording medium.

[0072] The CPU 2212 may also cause all or a necessary portion of a file or database stored on an external recording medium such as the hard disk drive 2224, the DVD-ROM drive 2226 (DVD-ROM 2201), an IC card, etc. to be read into the RAM 2214, and perform various types of processing on the data on the RAM 2214. The CPU 2212 then writes back the processed data to the external recording medium.

[0073] Various types of information, such as various types of programs, data, tables, and databases, may be stored on the recording medium and may undergo information processing. The CPU 2212 may perform various types of processing on data read from the RAM 2214, including various types of operations, information processing, conditional judgment, conditional branching, unconditional branching, information search / replacement, etc., as described throughout this disclosure and specified by the instruction sequences of the programs, and write the results back to the RAM 2214. The CPU 2212 may also search for information in a file, database, etc. on the recording medium. For example, if multiple entries each having an attribute value of a first attribute associated with an attribute value of a second attribute are stored on the recording medium, the CPU 2212 may search for an entry that matches a condition specified by the attribute value of the first attribute from among the multiple entries, read the attribute value of the second attribute stored in the entry, and thereby obtain the attribute value of the second attribute associated with the first attribute that satisfies a predetermined condition.

[0074] The above-described programs or software modules may be stored in a computer-readable medium on or near the computer 2200. A recording medium such as a hard disk or RAM provided in a server system connected to a dedicated communication network or the Internet can also be used as a computer-readable medium, thereby providing the programs to the computer 2200 via the network.

[0075] Although the present invention has been described above using embodiments, the technical scope of the present invention is not limited to the scope described in the above embodiments. It will be apparent to those skilled in the art that various modifications and improvements can be made to the above embodiments. It is clear from the claims that such modifications and improvements can also be included within the technical scope of the present invention.

[0076] It should be noted that the execution order of each process, such as operations, procedures, steps, and stages, in the devices, systems, programs, and methods shown in the claims, specifications, and drawings is not specifically stated as "before," "prior to," etc., and that the processes can be performed in any order unless the output of a previous process is used in a subsequent process. Even if the operational flow in the claims, specifications, and drawings is described using "first," "next," etc. for convenience, this does not mean that the processes must be performed in this order. [Explanation of symbols]

[0077] 100 Microfluidic device, 100a top surface, 100b bottom surface, 100c side surface, 100d side surface, 100e side surface, 100f side surface, 101 structure, 101a top surface, 102 transmission surface, 110 Microfluidic device, 121 transmission window, 122 positioning marker, 123 transmission window, 124 positioning marker, 131 transmission window, 132 convex portion, 133 positioning marker, 141 transmission window, 142 concave portion, 143 positioning marker, 144 optical scanner, 151 transmission window, 152 concave portion, 152a inclined plane, 152b curved surface, 153 positioning marker, 154 optical scanner, 161 transmission window, 162 mask, 163 positioning marker, 200 observation device, 210 Sheet illumination optical system, 211 optical fiber, 212 collector lens, 213 cylindrical lens, 214 variable aperture, 220 observation optical system, 221 objective lens, 222 two-dimensional detector, 230 stage, 250 IR illumination light source, 260 marker observation optical system, 261 IR detector, 262 lens, 263 dichroic mirror, 270 observation device, 300 observation area, 301 optical fiber, 2200 computer, 2201 DVD-ROM, 2210 host controller, 2212 CPU, 2214 RAM, 2216 graphics controller, 2218 display device, 2220 input / output controller, 2222 communication interface, 2224 hard disk drive, 2226 DVD-ROM drive, 2230 ROM, 2240 input / output chip, 2242 keyboard

Claims

1. 1. A microfluidic device having a top surface and a bottom surface and a plurality of side surfaces, At least one of the plurality of side surfaces has an optically transmissive portion that transmits illumination light from outside. Microfluidic devices.

2. The microfluidic device according to claim 1 , wherein the optically transmissive portion is formed corresponding to an observation area to be observed.

3. The microfluidic device according to claim 1 , wherein a plurality of the optically transmissive portions are formed on the at least one side surface.

4. The microfluidic device according to claim 1 , wherein the one side surface has a marker for locating or identifying the position of the optically transmissive portion.

5. The microfluidic device of claim 1 , wherein the optically transmissive portion has an optical function.

6. The microfluidic device of claim 5 , wherein the optical function is a convex lens function.

7. The microfluidic device of claim 5 , wherein the optical function is a concave lens function.

8. The microfluidic device according to claim 5 , wherein the optical function is a function of splitting the illumination light into multiple directions.

9. The microfluidic device of claim 1 , further comprising a mask formed in the optically transmissive portion, the mask blocking a portion of the illumination light.

10. 10. The microfluidic device of claim 1, wherein at least one of the top surface and the bottom surface has an optically transmissive portion that transmits output light output from within the microfluidic device.

11. An observation device for a microfluidic device, an illumination optical system for illuminating a microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, at least one of which has an optically transmissive portion through which illumination light from an external source passes, with the illumination light irradiating the microfluidic device from the optically transmissive portion; an observation optical system that receives output light from at least one of the top surface and the bottom surface; Observation device for microfluidic devices.

12. The observation apparatus for a microfluidic device according to claim 11 , wherein the angle of incidence of the illumination light on the optically transmitting portion is variable.

13. the one side has a marker for locating or identifying the position of the optically transmissive portion; The microfluidic device observation apparatus according to claim 11 or 12, further comprising a marker observation optical system for observing the marker, and a light source for irradiating the marker with illumination light.

14. Further comprising a control device; The microfluidic device observation apparatus according to claim 11 , wherein the control device controls the illumination optical system to form the illumination light based on information relating to an observation region corresponding to the optically transmitting portion.

15. A method for observing a microfluidic device, comprising: A microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, at least one of the plurality of side surfaces having an optically transparent portion that transmits illumination light from outside, an illumination step of illuminating the illumination light from the optically transmissive portion; and receiving output light from at least one of the top surface and the bottom surface.

16. The method for observing a microfluidic device according to claim 15 , wherein the angle of incidence of the illumination light on the optically transmitting portion is variable.

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