WO2024024697A1 - Dispositif microfluidique, appareil et procédé d'observation pour dispositif microfluidique - Google Patents

Dispositif microfluidique, appareil et procédé d'observation pour dispositif microfluidique Download PDF

Info

Publication number
WO2024024697A1
WO2024024697A1 PCT/JP2023/026894 JP2023026894W WO2024024697A1 WO 2024024697 A1 WO2024024697 A1 WO 2024024697A1 JP 2023026894 W JP2023026894 W JP 2023026894W WO 2024024697 A1 WO2024024697 A1 WO 2024024697A1
Authority
WO
WIPO (PCT)
Prior art keywords
microfluidic device
observation
optically transparent
transparent portion
light
Prior art date
Application number
PCT/JP2023/026894
Other languages
English (en)
Japanese (ja)
Inventor
久美子 松爲
Original Assignee
株式会社ニコン
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社ニコン filed Critical 株式会社ニコン
Publication of WO2024024697A1 publication Critical patent/WO2024024697A1/fr

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence

Definitions

  • the present invention relates to a microfluidic device, a microfluidic device observation apparatus, and a microfluidic device observation method.
  • Patent Document 1 describes a microfluidic device observation apparatus and a microfluidic device observation method for observing a specimen existing inside the channels of a microfluidic device in which one or more channels are arranged. Are listed. [Prior art documents] [Patent document] [Patent Document 1] International Publication No. 2020/021604 [General Disclosure]
  • a microfluidic device having a top surface, a bottom surface, and a plurality of side surfaces, wherein at least one of the plurality of side surfaces is an optical device that transmits illumination light from the outside.
  • a microfluidic device is provided having a permeable portion.
  • the optically transparent portion may be formed corresponding to an observation area to be observed.
  • a plurality of the optically transparent parts may be formed on the at least one side surface.
  • the one side surface may include a marker for searching or specifying the position of the optically transparent portion.
  • the optically transparent portion may have an optical function.
  • the optical function may be a convex lens function.
  • the optical function may be a concave lens function.
  • the optical function may be a function of separating the illumination light into multiple directions.
  • a mask may be formed on the optically transparent portion to block part of the illumination light.
  • the optically transparent portion may have a mark formed corresponding to an observation area to be observed.
  • At least one of the top surface and the bottom surface may have an optically transparent portion that transmits output light output from within the microfluidic device.
  • an observation apparatus for a microfluidic device which has a top surface, a bottom surface, and a plurality of side surfaces, and at least one of the plurality of side surfaces receives illumination light from the outside.
  • An illumination optical system that illuminates illumination light from the optically transparent portion, and an observation optical system that receives output light from at least one of the top surface and the bottom surface, are provided for a microfluidic device having an optically transparent portion that transmits the microfluidic device.
  • an observation apparatus for a microfluidic device is provided.
  • the angle of incidence of the illumination light onto the optically transparent portion may be variable.
  • the one side may include a marker for searching or specifying the position of the optically transparent portion, and may include a marker observation optical system for observing the marker, and a light source for irradiating the marker with illumination light. It may further include.
  • control device may control the illumination optical system to form the illumination light based on information regarding the observation area corresponding to the optically transparent portion.
  • a method for observing a microfluidic device which has a top surface, a bottom surface, and a plurality of side surfaces, and at least one of the plurality of side surfaces is illuminated by external illumination light.
  • a microfluidic device having an optically transparent portion that transmits the microfluidic device, an illumination stage that illuminates the illumination light from the optically transparent portion, and an observation stage that receives output light from at least one of the top surface and the bottom surface;
  • a method for observing a microfluidic device is provided.
  • the angle of incidence of the illumination light onto the optically transparent portion may be variable.
  • FIG. 1 is a perspective view showing an example of a schematic configuration of a microfluidic device 100 in a first embodiment.
  • FIG. 1 is a side view showing an example of a schematic configuration of a microfluidic device 100 in a first embodiment.
  • An example of a schematic configuration of an observation device 200 of a microfluidic device 100 in the first embodiment is shown.
  • FIG. 3 is an explanatory diagram of angle adjustment of sheet light L1 of the microfluidic device 100 in the first embodiment. It is a flowchart showing the operation of the observation device 200 of the microfluidic device 100 in the first embodiment.
  • FIG. 2 is a perspective view showing an example of a schematic configuration of a microfluidic device 110 in a second embodiment.
  • FIG. 3 is a side view showing an example of a schematic configuration of a microfluidic device 110 in a second embodiment.
  • An example of a schematic configuration of an observation device 270 of the microfluidic device 110 in the second embodiment is shown. It is a flowchart showing the operation of the observation device 270 of the microfluidic device 110 in the second embodiment.
  • a transmission window 131 in a first modification is shown.
  • a transmission window 141 in a second modification is shown.
  • a transmission window 151 in a third modification is shown.
  • a transmission window 161 in a fourth modification is shown. It is a figure which shows the modification of an illumination method.
  • An example of a computer 2200 is shown.
  • FIG. 1 is a perspective view showing an example of a schematic configuration of a microfluidic device 100 in the first embodiment.
  • microfluidic devices 100 have a top surface 100a, a bottom surface 100b, and four side surfaces 100c to 100f.
  • the top surface 100a is a surface facing in the direction opposite to the direction of gravity when the microfluidic device 100 is placed for observation
  • the bottom surface 100b is a surface facing in the direction opposite to the direction of gravity when the microfluidic device 100 is placed for observation.
  • the side facing The four side surfaces 100c to 100f are surfaces perpendicular to the top surface 100a and the bottom surface 100b.
  • the side surface 100c is a side surface of the microfluidic device 100 facing in the -X direction
  • the side surface 100d is a side surface facing the +Y direction of the microfluidic device 100
  • the side surface 100e is a side surface facing the +X direction of the microfluidic device 100.
  • the side face 100f is the side face of the microfluidic device 100 facing in the -Y direction.
  • the microfluidic device 100 has a plurality of layers, and each layer of the microfluidic device 100 has a plurality of structures 101 such as microchannels and injection ports through which cells can be injected.
  • 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.
  • the microfluidic device 100 has a transmission surface 102 on which at least one of the plurality of sides is an optically transparent portion that transmits illumination light from the outside.
  • the illumination light is, for example, excitation light in the visible or infrared region for fluorescence observation, and illumination light in the visible region for observation by transmission or reflection.
  • the top surface 101a or bottom surface 100b of the microfluidic device 100 has an optically transparent portion that transmits observation light.
  • the observation light is, for example, fluorescence in the visible range for fluorescence observation, and observation light in the visible range for observation by transmission or reflection.
  • the microfluidic device 100 is a microfluidic device in which samples corresponding to minute biological substances such as DNA, proteins, cells, cell masses (spheroids, organoids, etc.), tissues, etc. are placed on a small substrate to detect genetic defects, protein distribution, and reactions.
  • the microfluidic device 100 in this embodiment includes an organ chip (organ on a chip), a biological function chip (organ on a chip), an MPS (microphysical systems), a biochip (biochip), a microfluidic chip (mic rofluidic chip) , microchip, cell culture chip, microchannel chip, etc.
  • the microfluidic device 100 is used, for example, for culturing and analyzing cells and tissues. Furthermore, chemical substances (drugs) are added to the mixture and used for reaction evaluation or analysis with cultured cells.
  • the microfluidic device 100 may include both a device in which organ cells are cultured to express a biological function and an "empty" device body in which organ cells are not yet cultured.
  • the microfluidic device 100 can be manufactured using, for example, stereolithography three-dimensional printing technology and a solution cast molding process. In addition to the above techniques, the microfluidic device 100 can be manufactured using other microfabrication techniques such as MEMS (Micro Electro Mechanical Systems).
  • MEMS Micro Electro Mechanical Systems
  • Each layer of the microfluidic device 100 is composed of, for example, a substrate.
  • the substrate may be made of glass, for example.
  • the substrate may be made of a resin material such as polymethyl methacrylate (PMMA), polycarbonate (PC), cycloolefin copolymer (COC), cycloolefin polymer (COP), polystyrene (PS), silicone, or the like.
  • Microfluidic device 100 may have a cover that covers the entire microfluidic device 100.
  • FIG. 2 is a side view showing an example of a schematic configuration of the microfluidic device 100 in the first embodiment.
  • FIG. 2 is a diagram of the microfluidic device 100 in FIG. 1 viewed from the ⁇ X direction, and shows a side surface 100c of the microfluidic device 100.
  • the entire side surface 100c is a transmitting surface 102 that transmits illumination light from the outside.
  • External illumination light is light that illuminates the microfluidic device 100 to illuminate the microfluidic device 100 when observing the microfluidic device 100 with an observation device such as a microscope. Therefore, the transmission surface 102 transmits illumination light from the outside.
  • the transmitting surface 102 is made of a material that transmits illumination light from the outside, and is made of glass, for example.
  • the side surface 100c which is the transmission surface 102, may have a mark formed corresponding to an observation area to be observed. For example, although not shown in FIG. 2, if there is an observation area 300 as shown in FIG. may be formed. This facilitates searching and specifying the position of the observation area.
  • FIG. 3 shows an example of a schematic configuration of the observation device 200 of the microfluidic device 100 in the first embodiment.
  • the observation device 200 in this embodiment is an inverted microscope, for example. However, an upright microscope is also applicable.
  • the observation device 200 acquires an image of the observation device 200 by illuminating the microfluidic device 100 with sheet light L1, which is excitation light, and capturing fluorescence excited from the fluorescent dye of the observation device 200.
  • the sheet light L1 is illumination light having a sheet-shaped illumination area.
  • the observation device 200 includes 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 arranged on a stage 230 of the observation apparatus 200, and by moving the stage 230 in the vertical direction and the horizontal direction (XYZ direction), the microfluidic device 100 is moved in the vertical direction and the horizontal direction (XYZ direction). It is movable.
  • the sheet illumination optical system 210, observation optical system 220, and stage 230 are connected to the PC 240 and can be automatically controlled.
  • 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 arranged on the -X direction side of the side surface of the microfluidic device 100, and illuminates the sheet light L1 toward the microfluidic device 100.
  • the output end face of the optical fiber 211 which is 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 light L1 having a sheet-like shape 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 sheet light L1 has a longitudinal direction in a direction perpendicular to the optical axis of the sheet illumination optical system 210.
  • the observation area 300 is an area of the microfluidic device 100 that the user wants to observe with a microscope.
  • the sheet light L1 enters the inside of 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.
  • sheet light L1 is illuminated toward the microfluidic device 100 in parallel to the X axis.
  • 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.
  • the illumination area of the sheet light L1 can be configured to be movable in the height direction ( ⁇ Z direction). It is desirable that the intensity and longitudinal width of the sheet light L1 be automatically finely adjusted so as to minimize ghosts caused by diffused reflection on the structures 101 inside the microfluidic device 100.
  • the observation optical system 220 includes 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 in which low magnification/medium/high magnification can be electrically switched.
  • the sheet light L1 emitted from the sheet illumination optical system 210 illuminates cells in the observation region 300 of the microfluidic device 100.
  • the observation area 300 is not limited to cells, and may include cell clusters and tissues.
  • the cells in the observation area 300 are excited by the sheet light L1 and emit fluorescence L2 as output light. 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. Note that the observation may be performed from the direction of the top surface 100a of the microfluidic device 100, in which case the observation optical system 220 is arranged in the direction of the top surface 100a of the microfluidic device 100, and the top surface 100a is made of a material that transmits the fluorescence L2. .
  • the fluorescence L2 emitted from the bottom surface 100b of the microfluidic device 100 is focused by the objective lens 221 and detected by the 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 and a dichroic mirror.
  • the PC (control device) 240 includes a CPU and a memory (storage unit), and the CPU controls the operation of the observation device 200 by reading and executing a control program stored in the memory. As shown by the dashed line in FIG. 3, the PC 240 is connected to the stage 230 of the microscope, etc., and can control the operation of the entire microscope.
  • the PC 240 includes an input unit that receives various instructions and settings from the user and sends them to the control unit of the PC 240, and a GUI (Graphical User Interface) screen and a It has a display section that displays various dialogs and the like.
  • FIG. 4 is an explanatory diagram of the angle adjustment of the sheet light L1 of the microfluidic device 100 in the first embodiment.
  • FIG. 4 is a partially enlarged view of the observation device 200, and only some of the components of the observation device 200 are illustrated.
  • the sheet light L1 entering the microfluidic device 100 from the sheet illumination optical system 210 was parallel to the X axis.
  • FIG. 4 is an explanatory diagram of the angle adjustment of the sheet light L1 of the microfluidic device 100 in the first embodiment.
  • FIG. 4 is a partially enlarged view of the observation device 200, and only some of the components of the observation device 200 are illustrated.
  • the sheet light L1 entering the microfluidic device 100 from the sheet illumination optical system 210 was parallel to the X axis.
  • the output end of the optical fiber 211 faces diagonally upward, and the sheet light L1 illuminates the observation area 300 at a diagonally downward angle by passing through the collector lens 212 and the cylindrical lens 213. ing.
  • the observation area 300 can be illuminated from an appropriate angle, for example.
  • the observation area 300 can be illuminated while avoiding the obstacle. Adjustment of the direction (tilt) of the output end of the optical fiber 211 and adjustment of the height position (shift) (position in the Z direction) of the output end of the optical fiber 211 can be performed in combination.
  • FIG. 5 is a flowchart showing the operation of the observation apparatus 200 of the microfluidic device 100 in the first embodiment.
  • step S01 the user places the microfluidic device 100 on the stage 230 of the microscope.
  • step S02 the user selects the type of microfluidic device 100 on the GUI.
  • step S03 based on the type of microfluidic device 100 selected by the user, the PC 240 determines whether or not it has information on the microfluidic device 100 of the selected type.
  • the information on the microfluidic device 100 provides at least one of the shape, structure, size, manufacturer, cultured cell information, and model number of the microfluidic device 100, for example. If the PC 240 has information on the microfluidic device 100 (step S03: YES), the process advances to step S05, reads out the information on the microfluidic device 100, and allows the user to select the relevant one from among multiple assays displayed on the GUI. Select an assay that is compatible with microfluidic device 100.
  • step S03 If the PC 240 does not have information about the microfluidic device 100 (step S03: NO), the process proceeds to step S04, and 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 obtains a predetermined result by acquiring an image and performing predetermined analysis and evaluation processing on the obtained cell image. There are calculations, cell viability determination, etc.
  • 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 and moves the field of view of the objective lens 221 to the first observation area 300.
  • the position of the observation area 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 based on the specified position, the process of step S07 is performed by the observation apparatus 200. done automatically.
  • the process in step S07 may be performed manually by the user.
  • step S08 the observation conditions (including illumination conditions, magnification of the objective lens 221, relative position between the objective lens 221 and the observation area 300, etc.) are changed in order to acquire an image of the observation area 300, and the observation area 300 is Get the image of.
  • the user refers to the three-dimensional image of the entire microfluidic device 100 displayed on the GUI, takes into consideration the structure 101, and selects the optimal transmission window for the first observation area 300. Set the lighting conditions and irradiate the sheet light. Note that by linking information regarding the observation area 300, information regarding the structure 101, and information regarding the transmission window and storing it in the PC 240, the user can perform the first observation simply by specifying the observation area 300.
  • the transmission window corresponding to the region 300 may be irradiated with sheet light under optimal illumination conditions.
  • the illumination conditions include, for example, the intensity of the sheet light, the width in the longitudinal direction of the sheet light, the illumination position, the illumination angle, the illumination wavelength, the size of a variable aperture corresponding to the longitudinal width of the sheet light, and the like.
  • 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.
  • step S10 it is determined whether there is a next observation area. If there is another observation area (step S10: YES), the process returns to step S07 and the processes from steps S07 to S09 are repeated. If there is no next observation area (step S10: NO), the process proceeds to the next steps S11 and S12.
  • step S11 assay image processing (image analysis) is performed, and in step S12, the processing results (analysis results) are saved.
  • the side surface 100c of the microfluidic device 100 has the transmission surface 102, which is an optically transparent portion that transmits illumination light from the outside.
  • the sheet light L1 for observing the microfluidic device 100 can enter the inside of the microfluidic device 100 from the side surface 100c of the microfluidic device 100. Therefore, for example, even if a structure 101 such as a microchannel is installed on the top surface 101a of the microfluidic device 100 and it is difficult to perform transmission observation of the desired observation area 300 from the top, the sheet light L1 is transmitted from the side surface 100c. can be observed by making it incident.
  • the internal structures 101 of the microfluidic device 100 overlap in the height direction, making it difficult to pinpoint the desired observation area 300 from above, or the microfluidic device 100 has many layers. Even if it is difficult to transparently observe the desired observation area 300 from the top because it is thick in the height direction, the sheet light L1 can be incident on the side surface 100c to perform observation.
  • the incident angle of the sheet light L1 to the transmission window for observing the microfluidic device 100 can be adjusted.
  • the observation region 300 can be observed with good contrast while avoiding the structures 101 such as microchannels in the microfluidic device 100.
  • the same effects as the microfluidic device 100 in the first embodiment can be achieved.
  • FIG. 6 is a perspective view showing an example of a schematic configuration of the microfluidic device 110 in the second embodiment.
  • the same components as the microfluidic device 100 of the first embodiment will be given the same reference numerals and the explanation will be omitted.
  • a transmission window 121 and a transmission window 123 which are optical transmission portions that transmit illumination light from the outside, are formed on a side surface 100c.
  • FIG. 7 is a side view showing an example of a schematic configuration of the microfluidic device 110 in the second embodiment.
  • FIG. 7 is a diagram of the microfluidic device 110 of FIG. 6 viewed from the ⁇ X direction, and shows 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 in the second embodiment.
  • the transmission window 121 is provided at the lower left of the side surface 100c, and the transmission window 123 is provided at the upper right of the side surface 100c.
  • the transmission window 121 and the transmission window 123 are provided corresponding to the position of the region to be observed in the microfluidic device 110.
  • the transmission window 121 and the transmission window 123 are made of a material that transmits illumination light from the outside for observation of the microfluidic device 110, and are made of glass, for example.
  • the transmission window 121 and the transmission window 123 in the second embodiment have a flat shape with a constant thickness in the X direction.
  • a positioning marker 122 and a positioning marker 124 are formed near the transparent window 121 and the transparent window 123 (for example, near the four corners).
  • the vicinity of the transmission window refers to the outside or inside of the transmission window. It is not limited to the vicinity of the four corners, but may be two corners on the diagonal line.
  • the positioning marker 122 and the positioning marker 124 are used to search and specify the positions of the transparent window 121 and the transparent window 123.
  • the positioning marker 122 and the positioning marker 124 may be formed of a material (metal deposited film, dielectric multilayer film) that has a reflective property for IR (infrared) light emitted from an IR illumination light source 250, which will be described later. Alternatively, it may be made of a material that scatters IR light, or it may be processed with frosted glass that scatters IR light.
  • the positioning marker 122 and the positioning marker 124 are different in shape, color, or material.
  • FIG. 8 shows an example of a schematic configuration of the observation device 270 of the microfluidic device 110 in the second embodiment.
  • 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.
  • the microfluidic device 110 in the second embodiment further includes an IR illumination light source 250 and a marker observation optical system 260, compared to the microfluidic device 100 in the first embodiment.
  • the IR illumination light source 250 and the marker observation optical system 260 are used to confirm the illumination mode of the sheet light L1 from the sheet illumination optical system 210.
  • the IR illumination light source 250 illuminates IR light toward at least one of the transmission window 121 and the transmission window 123 in which the positioning marker 122 and the positioning marker 124 are provided.
  • IR illumination source 250 includes, for example, at least one infrared light emitting diode (LED).
  • LED infrared light emitting diode
  • IR illumination source 250 may include multiple LEDs. The IR light is reflected or partially absorbed by at least one of the positioning marker 122 of the transmission window 121 and the positioning marker 124 of the transmission window 123 illuminated with the IR light, and enters the marker observation optical system 260.
  • the marker observation optical system 260 includes an IR detector 261, a lens 262, and a dichroic mirror 263.
  • the dichroic mirror 263 has a characteristic of transmitting the sheet light L1 and reflecting the IR light.
  • the range that can be detected by the marker observation optical system 260 at once is the range that includes 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 incident on the marker observation optical system 260 is reflected by the dichroic mirror 263, focused by the lens 262, and incident on the IR detector 261.
  • Each of the positioning marker 122 and the positioning marker 124 is composed of a plurality of members, but since the plurality of members can be illuminated and observed at once, the respective positions of the transmission window 121 and the transmission window 123 can be adjusted. can be quickly identified.
  • the position of the transmission window 121 can be specified. Therefore, for example, if the sheet light L1 travels on the optical axis of the sheet illumination optical system 210 and is incident on the ⁇ Z direction side with respect to the position of the transmission window 121, the desired observation area 300 cannot be illuminated. To do this, the sheet light L1 is transmitted 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. The desired observation area 300 can be illuminated using the same method.
  • FIG. 9 is a flowchart showing the operation of the observation device 270 of the microfluidic device 110 in the second embodiment.
  • the operations of steps S01 to S12 of the observation device 270 in the second embodiment are the same as the operations of steps S01 to S12 of the observation device 200 in the first embodiment.
  • the observation device 270 in the second embodiment further includes operations in steps S13 and S14.
  • the marker observation optical system 260 detects a positioning marker (for example, positioning marker 122) on a transmission window (for example, transmission window 121). By detecting the positioning marker 122, the position of the transmission window 121 can be specified.
  • optimal illumination conditions are set based on information regarding the structure 101 linked to the identified transmission window 121 and information regarding the observation area 300.
  • the illumination conditions include, for example, the intensity of the sheet light, the width in the longitudinal direction of the sheet light, the illumination position, the illumination angle, the illumination wavelength, the size of a variable aperture corresponding to the longitudinal width of the sheet light, and the like.
  • At least one of the transmission windows 121 and 123 may have an optical function of condensing the sheet light L1 or controlling the traveling direction of the sheet light L1.
  • FIG. 10 shows a transmission window 131 in a first modification.
  • FIG. 10 shows a cross-sectional view of the side surface 100c.
  • a convex portion 132 having a convex lens function is integrally formed in the transmission window 131 in the first modification, so that the transmission window 131 has a convex lens function as an example of an optical function. Since the transmission window 131 has a convex lens function, it is 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.
  • FIG. 11 shows a transmission window 141 in a second modification.
  • 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.
  • a concave portion 142 having a concave lens function is integrally formed in the transmission window 141 in the second modification, and thus the transmission window 141 has a concave lens function as an example of an optical function.
  • An optical scanner 144 that scans the sheet light L1 incident on the transmission window 141 is arranged on the ⁇ X direction side of the transmission window 141, and can scan the sheet light L1 and deflect the sheet light L1 in multiple directions. can.
  • the transmission window 141 has a concave lens function, for example, when the sheet light L1 is scanned by the optical scanner 144, the light beam is not refracted at the interface, so that the incident angle of the sheet light L1 can be changed as scanned, and This has the effect of keeping the thickness of the sheet light L1 constant. Further, when it is desired to illuminate with a wide range of illumination light, the light beam of the sheet light L1 can be expanded. Positioning markers 143 are formed at the four corners of the transmission window 141.
  • FIG. 12 shows a transmission window 151 in a third modification.
  • the transmission window 151 in the third modification is integrally formed with a concave portion 152 consisting of an inclined plane 152a and a curved surface 152b that are inclined with respect to the side surface.
  • the inclined plane 152a has a function of allowing illumination light from a specific direction to enter without being refracted.
  • the curved surface 152b has a concave lens function as an example of an optical function.
  • An optical scanner 154 that scans the sheet light L1 incident on the transmission window 151 is arranged on the ⁇ X direction side of the transmission window 151, and can change the incident angle by scanning the sheet light L1. Since the curved surface 152b of the transmission window 151 has a concave lens function, it can have the same function as the transmission window 141 in the second modification. Further, a plurality of observation areas 300 can be collectively illuminated from the transmission window 151 at a desired angle. Positioning markers 153 are formed at the four corners of the transmission window 151.
  • FIG. 13 shows a transmission window 161 in a fourth modification.
  • FIG. 13 shows a cross-sectional view of the side surface 100c.
  • a plurality of masks 162 are formed in the transmission window 161 in the fourth modification to block part of the sheet light L1 (in a direction that is not desired to be irradiated).
  • a plurality of observation regions 300 can be collectively illuminated.
  • Positioning markers 163 are formed at the four corners of the transmission window 161.
  • the transmission windows 131 to 161 shown in FIGS. 10 to 13 may be combined arbitrarily.
  • the mask 162 may be combined with the transmission window 141 having a concave lens function.
  • the mask 162 may be combined with the transmission window 151 having a plurality of inclined planes 152a and curved surfaces 152b.
  • microfluidic device 110 in the second embodiment the same effects as the microfluidic device 100 in the first embodiment can be achieved.
  • a plurality of transmission windows 121 and transmission windows 123 are formed on the side surface 101c. This makes it easy to observe multiple observation areas 300 at different positions.
  • the entire side surface 101c is not a transmission surface, but a plurality of transmission windows 121 and transmission windows 123 are formed in a part of the side surface 101c.
  • the other parts of the side surface 101c can be made of any material, and material costs can be reduced.
  • the four corners of the transmission window 121 and the transmission window 123 are provided with positioning markers 122 and positioning markers for specifying the positions of the transmission window 121 and the transmission window 123.
  • 124 is formed, and the observation device 270 further includes an IR illumination light source 250 and a marker observation optical system 260 for detecting the positioning marker 122 and the positioning marker 124.
  • the transmission windows 131 to 161 of the first modification to the fourth modification have various optical functions.
  • the sheet light L1 from the sheet illumination optical system 210 can be optically manipulated.
  • the transmitting surface 102 of the first embodiment may be provided with an optical function.
  • the same effects as the microfluidic device 110 in the second embodiment can be achieved.
  • an optically transparent portion was provided on the side surface 101c of the microfluidic devices 100, 110, and the observation region 300 was irradiated from the side surface 101c.
  • a configuration may also be adopted in which a reflection mirror is provided inside the microfluidic devices 100 and 110 so that the direction of the sheet light L1 irradiated from the side surface 101c can be changed.
  • the sheet light L1 irradiated from the side surface 101c is light that travels in a direction generally parallel to the XY plane
  • the sheet light L1 irradiated from the side surface 101c may be reflected by a reflective mirror and directed toward the Z direction.
  • an optically transparent portion was provided on the side surface 101c of the microfluidic device 100, 110, and the inside was observed by irradiating the sheet light L1 from the side surface 101c.
  • the sheet light L1 may be further irradiated from the top surface 101a or the bottom surface 101b of the microfluidic device 100, 110, or the sheet light L1 may be irradiated from the side surface 101c and the top surface 101a or the bottom surface 101b may be combined. good.
  • 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 is actively lowered than on the side surface 101c on which the transmission surface and the transmission window are formed.
  • the side surface may be rubbed to make it easier to grip with human hands.
  • the side surface may be covered with a light-shielding mask so that external light from the side surface does not affect internal cells.
  • two transmission windows 121 and 123 were formed on one side surface 100c of the microfluidic device 110.
  • the number of transmission windows formed on one side surface may be three or more.
  • a plurality of transmission windows may be formed on each of the plurality of side surfaces. In that case, it is desirable to provide the sheet illumination optical system 210 corresponding to the plurality of side surfaces on which the transmission windows are formed.
  • a transmission window was formed on the side surface 100c of the microfluidic device 110, and positioning markers were formed at the four corners of the transmission window.
  • the positioning marker may be formed inside the microfluidic device 110 instead of being formed near the transmission window.
  • the markers may be formed at parts other than the four corners of the transmission window. The number of markers to be formed does not have to be four.
  • FIG. 14 is a diagram showing a modification of the lighting method.
  • the sheet illumination optical system 210 was provided in the side direction of the microfluidic devices 100, 110, and the sheet light L1 was irradiated from the side of the microfluidic devices 100, 110 to observe the inside.
  • an optical fiber 301 that emits light entirely or a light source unit in which a plurality of LEDs (not shown) are arranged in a straight line is installed on the side of the microfluidic device 100, 110.
  • the interior may be illuminated by arranging them parallel to each other (in the XY plane direction).
  • 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. By adjusting the Z position, dark field illumination and bright field illumination can be realized.
  • a computer-readable medium may include any tangible device capable of storing instructions for execution by a suitable device, such that the computer-readable medium having instructions stored thereon is illustrated in a flowchart or block diagram.
  • An article of manufacture will be provided that includes instructions that can be executed to create a means for performing the operations.
  • Examples of computer readable media may include electronic storage media, magnetic storage media, optical storage media, electromagnetic storage media, semiconductor storage media, and the like.
  • Computer readable media 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 Disk (DVD), Blu-ray (RTM) Disc, Memory Stick, Integrated Circuit cards etc. may be included.
  • RAM random access memory
  • ROM read only memory
  • EPROM or flash memory erasable programmable read only memory
  • EEPROM Electrically Erasable Programmable Read Only Memory
  • SRAM Static Random Access Memory
  • CD-ROM Compact Disc Read Only Memory
  • DVD Digital Versatile Disk
  • RTM Blu-ray
  • Memory Stick Integrated Circuit cards etc.
  • Computer-readable instructions may include assembler instructions, Instruction Set Architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state configuration data, or instructions such as Smalltalk®, JAVA®, C++, etc. any source code or object code written in any combination of one or more programming languages, including object-oriented programming languages and traditional procedural programming languages, such as the "C" programming language or similar programming languages; may include.
  • ISA Instruction Set Architecture
  • Computer-readable instructions may be implemented on a processor or programmable circuit of a general purpose computer, special purpose computer, or other programmable data processing device, either locally or over a wide area network (WAN), such as a local area network (LAN), the Internet, etc. ), computer-readable instructions may be executed to create a means for performing the operations specified in the flowchart or block diagrams.
  • processors include computer processors, processing units, microprocessors, digital signal processors, controllers, microcontrollers, and the like.
  • FIG. 15 illustrates an example computer 2200 in which aspects of the invention may be implemented, in whole or in part.
  • a program installed on computer 2200 may cause computer 2200 to function as an operation or one or more sections of an apparatus according to an embodiment of the present invention, or to perform one or more operations associated with an apparatus according to an embodiment of the present invention.
  • Sections and/or computer 2200 may be caused to perform a process or a step of a process according to an embodiment of the invention.
  • Such programs may be executed by CPU 2212 to cause computer 2200 to perform certain operations associated with some or all of the blocks in the flowcharts and block diagrams described herein.
  • the computer 2200 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.
  • 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 ROM 2230 and keyboard 2242, which are connected to input/output controller 2220 via input/output chip 2240.
  • the CPU 2212 operates according to programs stored in the ROM 2230 and RAM 2214, thereby controlling each unit.
  • Graphics controller 2216 obtains image data generated by CPU 2212, such as in a frame buffer provided in RAM 2214 or itself, and causes the image data to be displayed on display device 2218.
  • the communication interface 2222 communicates with other electronic devices via the network.
  • Hard disk drive 2224 stores programs and data used by CPU 2212 within computer 2200.
  • DVD-ROM drive 2226 reads programs or data from DVD-ROM 2201 and provides the programs or data to hard disk drive 2224 via RAM 2214.
  • the IC card drive reads programs and data from and/or writes programs and data to the IC card.
  • ROM 2230 stores therein programs such as a boot program executed by computer 2200 upon activation and/or programs dependent on the computer 2200 hardware.
  • Input/output chip 2240 may also connect various input/output units to input/output controller 2220 via parallel ports, serial ports, keyboard ports, mouse ports, etc.
  • a program is provided by a computer readable medium such as a DVD-ROM 2201 or an IC card.
  • the program is read from a computer readable medium, installed on hard disk drive 2224, RAM 2214, or ROM 2230, which are also examples of computer readable media, and executed by CPU 2212.
  • the information processing described in these programs is read by the computer 2200 and provides coordination between the programs and the various types of hardware resources described above.
  • An apparatus or method may be configured to implement the manipulation or processing of information according to the use of computer 2200.
  • the CPU 2212 executes a communication program loaded into the RAM 2214 and sends communication processing to the communication interface 2222 based on the processing written in the communication program. You can give orders.
  • the communication interface 2222 reads transmission data stored in a transmission buffer processing area provided in a recording medium such as a RAM 2214, a hard disk drive 2224, a DVD-ROM 2201, or an IC card under the control of the CPU 2212, and transmits the read transmission data. Data is transmitted to the network, or received data received from the network is written to a reception buffer processing area provided on the recording medium.
  • the CPU 2212 causes the RAM 2214 to read all or a necessary part of a file or database stored in an external recording medium such as a hard disk drive 2224, a DVD-ROM drive 2226 (DVD-ROM 2201), an IC card, etc. Various types of processing may be performed on data on RAM 2214. The CPU 2212 then writes back the processed data to the external recording medium.
  • an external recording medium such as a hard disk drive 2224, a DVD-ROM drive 2226 (DVD-ROM 2201), an IC card, etc.
  • Various types of processing may be performed on data on RAM 2214.
  • the CPU 2212 then writes back the processed data to the external recording medium.
  • the CPU 2212 performs various types of operations, information processing, conditional determination, conditional branching, unconditional branching, and information retrieval on the data read from the RAM 2214 as described elsewhere in this disclosure and specified by the instruction sequence of the program. Various types of processing may be performed, including /substitutions, etc., and the results are written back to RAM 2214. Further, the CPU 2212 may search for information in a file in a recording medium, a database, or the like.
  • the CPU 2212 search the plurality of entries for an entry that matches the condition, read the attribute value of the second attribute stored in the entry, and thereby associate it with the first attribute that satisfies the predetermined condition.
  • the attribute value of the second attribute may be acquired.
  • the programs or software modules described above may be stored on computer readable media on or near 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 be used as a computer-readable medium, thereby providing the program to the computer 2200 via the network. do.
  • 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 Ka, 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 Light 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, 2

Landscapes

  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un dispositif microfluidique ayant une surface supérieure, une surface inférieure et une pluralité de surfaces latérales, au moins une surface latérale de la pluralité de surfaces latérales comprenant une partie optiquement transparente qui transmet une lumière d'éclairage provenant de l'extérieur. L'invention concerne également un appareil d'observation pour dispositif microfluidique. L'appareil d'observation comprend : un système optique d'éclairage qui éclaire un dispositif microfluidique avec la lumière d'éclairage par l'intermédiaire de la partie optiquement transparente, le dispositif microfluidique ayant une surface supérieure, une surface inférieure et une pluralité de surfaces latérales, au moins une surface latérale de la pluralité de surfaces latérales comprenant une partie optiquement transparente qui transmet une lumière d'éclairage provenant de l'extérieur ; et un système optique d'observation qui reçoit la lumière de sortie provenant de la surface supérieure et/ou de la surface inférieure.
PCT/JP2023/026894 2022-07-27 2023-07-21 Dispositif microfluidique, appareil et procédé d'observation pour dispositif microfluidique WO2024024697A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2022-119218 2022-07-27
JP2022119218 2022-07-27

Publications (1)

Publication Number Publication Date
WO2024024697A1 true WO2024024697A1 (fr) 2024-02-01

Family

ID=89706527

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/026894 WO2024024697A1 (fr) 2022-07-27 2023-07-21 Dispositif microfluidique, appareil et procédé d'observation pour dispositif microfluidique

Country Status (1)

Country Link
WO (1) WO2024024697A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118150561A (zh) * 2024-03-29 2024-06-07 追光生物科技(深圳)有限公司 一种便携式的微流控观察和检测系统

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005188999A (ja) * 2003-12-24 2005-07-14 Matsushita Electric Ind Co Ltd 特定成分の濃度測定装置、特定成分の濃度測定方法
JP2018097001A (ja) * 2018-01-09 2018-06-21 株式会社東芝 光学センサ、分析装置および分析方法
WO2020085104A1 (fr) * 2018-10-24 2020-04-30 凸版印刷株式会社 Gobelet pour immunoessai, son procédé de production et procédé d'immunoessai

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005188999A (ja) * 2003-12-24 2005-07-14 Matsushita Electric Ind Co Ltd 特定成分の濃度測定装置、特定成分の濃度測定方法
JP2018097001A (ja) * 2018-01-09 2018-06-21 株式会社東芝 光学センサ、分析装置および分析方法
WO2020085104A1 (fr) * 2018-10-24 2020-04-30 凸版印刷株式会社 Gobelet pour immunoessai, son procédé de production et procédé d'immunoessai

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118150561A (zh) * 2024-03-29 2024-06-07 追光生物科技(深圳)有限公司 一种便携式的微流控观察和检测系统

Similar Documents

Publication Publication Date Title
CN103477197B (zh) 样品使用最大化的系统和方法
JP5087745B2 (ja) 複数の観察手法を用いた顕微鏡細胞観察・検査システム
WO2024024697A1 (fr) Dispositif microfluidique, appareil et procédé d'observation pour dispositif microfluidique
JP6925361B2 (ja) サンプル場所のための補助画像検出器を有する撮像システム
US9523640B2 (en) Method of fluorescent measurement of samples, and devices therefrom
CN102540445A (zh) 显微镜、区域判定方法和程序
JP6513802B2 (ja) ナノ粒子検出のためのレーザー光結合
CN109643012B (zh) 观察装置
CN114787684B (zh) 具有照明及收集物镜的非正交布置的开顶式光片显微镜
CN112114423A (zh) 一种便携式全自动多模态显微成像装置
Oheim High‐throughput microscopy must re‐invent the microscope rather than speed up its functions
JP6796379B2 (ja) レーザーマイクロダイセクター及びレーザーマイクロダイセクション方法
CN116802463A (zh) 具有共焦成像的用于微孔板的通用多功能检测系统
JP2012039929A (ja) 受精卵観察の画像処理方法、画像処理プログラム及び画像処理装置、並びに受精卵の製造方法
US20090292479A1 (en) Method for analyzing image from bio-detection analyzer
JP2022509823A (ja) 細胞培養監視のための小型光学結像システム
WO2024135236A1 (fr) Appareil d'observation pour dispositif microfluidique
JP2024016932A (ja) マイクロ流体デバイス、マイクロ流体デバイスの観察装置、マイクロ流体デバイスの観察方法
US6597522B2 (en) Optical system
WO2024122387A1 (fr) Dispositif de support et dispositif d'observation
US6384989B1 (en) Optical system
JP2024025864A (ja) マイクロ流体デバイス、マイクロ流体デバイス用観察装置、マイクロ流体デバイスの観察方法
JP6500371B2 (ja) 有形成分分析装置及び有形成分分析方法
US20220108064A1 (en) Experiment support apparatus, experiment support system, experiment support method, and computer-readable medium
WO2024080385A1 (fr) Dispositif, procédé et programme de traitement d'informations

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23846427

Country of ref document: EP

Kind code of ref document: A1