# US 9,435,734 B2 · Method for observing stem cells, method for removal of cell region in state tending toward differentiation, and device for observing stem cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,435,734 B2 - Document ID: US9435734B2 - Status: Granted - Kind: B2 - Application number: 14/806,999 - Filed: 2015-07-23 - Published: 2015-11-12 - Granted: 2016-09-06 - Earliest claimed: 2011-11-08 - Assignees: Hamamatsu Photonics K.K. - Applicants: Hamamatsu Photonics K.K. - Inventors: Norikazu Sugiyama; Takuji Kataoka; Ikuo Arata - Web page: https://patentgrep.com/patent/US9435734B2 ## CPC - G01N 21/47: Scattering, i.e. diffuse reflection (G01N21/25, G01N21/41 take precedence {G01N21/55 takes precedence}) - G01N 21/49: within a body or fluid - G01N 21/59: Transmissivity (G01N21/25 takes precedence) - G01N 33/4833: {of solid biological material, e.g. tissue samples, cell cultures (tissue in vivo A61B5/00; cell suspensions G01N33/48735)} - G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)} - G02B 21/088: {for both incident illumination and transillumination} - G01N 2201/0446: Multicell plate, sequential - G01N 2201/062: LED's - G02B 21/36: arranged for photographic purposes or projection purposes (G02B21/18 takes precedence){or digital imaging or video purposes including associated control and data processing arrangements (image data processing per se G06T)} - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS ## Abstract A method for observing stem cells by an observation device 1 comprises, placing stem cells C in a petri dish 11, mounting the petri dish 11 on a waveguide 21 via water 13, emitting illumination light L1 into the waveguide 21 and emitting the illumination light L1 to the stem cells C in the petri dish 11 via the water 13, and detecting scattered light L2, the scattered light L2 being the illumination light L1 emitted to the stem cells C that is scattered by the stem cells C and has passed through the waveguide 21. Then, in the light image detected by means of the scattered light L2, a region that is markedly darker than other regions is identified as being in the state tending toward differentiation. ## Claims ### Claim 1 A method for observing stem cells, comprising: irradiating stem cells placed on a transparent member with oblique illumination light; capturing a scattered light image of the stem cells by a camera having a two dimensional pixel structure, in which a plurality of pixels are two-dimensionally arranged; and identifying a stem cell that is in the state tending toward differentiation in the sterm cells based on the scattered light image, wherein the irradiating irradiates the stem cells with the oblique illumination light via the transparent member. ### Claim 2 (depends on 1) The method according to claim 1, wherein the identifying identifies a darker portion in the scattered light image of the stem cell that is in the state tending toward differentiation. ### Claim 3 (depends on 1) The method according to claim 1, wherein the capturing images light that is scattered by the stem cells and passed through the transparent member. ### Claim 4 (depends on 1) The method according to claim 1, further comprising, irradiating the stem cells placed on the transparent member with transmitted illumination light; and capturing a transmitted light image of the stem cells by the camera. ### Claim 5 (depends on 1) The method according to claim 1, further comprising, removing the identified stem cell from the stem cells. ### Claim 6 (depends on 1) The method according to claim 1, wherein the stem cells are iPS cells. ### Claim 7 An apparatus for observing stem cells, comprising: a light source configured to output light; an illumination optics configured to input the output light and irradiate stem cells placed on a transparent member with oblique illumination light; a camera having a two dimensional pixel structure, in which a plurality of pixels are two-dimensionally arranged and configured to capture a scattered light image of the stem cells; and a display configured to display the scattered light image, thereby identifying a stem cell that is in the state tending toward differentiation in the stem cells, wherein the illumination optics irradiates the stem cells with the oblique illumination light via the transparent member. ### Claim 8 (depends on 7) The apparatus according to claim 7, wherein the camera images light that is scattered by the stem cells and passed through the transparent member. ### Claim 9 (depends on 7) The apparatus according to claim 7, wherein the illumination optics comprises a light guide having a surface on which the transparent member can be mounted and another surface inputting the output light. ### Claim 10 (depends on 9) The apparatus according to claim 9, wherein the camera images light that is scattered by the stem cells and passed through the transparent member and the light guide. ### Claim 11 (depends on 7) The apparatus according to claim 7, wherein the stem cells are iPS cells. ### Claim 12 An apparatus for observing stem cells, comprising: a light source configured to output light; an illumination optics configured to input the output light and irradiate stem cells placed on a transparent member with oblique illumination light; a camera having a two dimensional pixel structure, in which a plurality of pixels are two-dimensionally arranged and configured to capture a scattered light image of the stem cells; and a display configured to display the scattered light image, thereby identifying a stem cell that is in the state tending toward differentiation in the stem cells, wherein the illumination optics comprises a light guide having a surface on which the transparent member can be mounted and another surface inputting the output light. ### Claim 13 (depends on 12) The apparatus according to claim 12, wherein the camera images light that is scattered by the stem cells and passed through the transparent member. ### Claim 14 (depends on 12) The apparatus according to claim 12, wherein the stem cells are iPS cells.