# US 10,481,094 B2 · Total internal reflection fluorescence imaging system and sequencing device > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,481,094 B2 - Document ID: US10481094B2 - Status: Granted - Kind: B2 - Application number: 15/752,417 - Filed: 2016-09-06 - Published: 2019-01-10 - Granted: 2019-11-19 - Earliest claimed: 2015-09-07 - Assignees: Genemind Biosciences Company Limited - Applicants: GeneMind Biosciences Company Limited - Inventors: Liangjin Ge; Luyang Zhao; Ping Wu; Qin Yan - Web page: https://patentgrep.com/patent/US10481094B2 ## CPC - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01N 21/648: {using evanescent coupling or surface plasmon coupling for the excitation of fluorescence} - G02B 21/0076: {arrangements using fluorescence or luminescence} - G02B 21/06: Means for illuminating specimens - G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} - G02B 21/18: Arrangements with more than one light path, e.g. for comparing two specimens - G02B 21/245: {using auxiliary sources, detectors} - G02B 27/141: {using dichroic mirrors} - G01N 2021/6419: {Excitation at two or more wavelengths} - 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 The present disclosure discloses a total internal reflection fluorescence imaging system and a sequencing device. The total internal reflection fluorescence imaging system includes a first imaging system and a second imaging system. The first imaging system includes N separate laser light paths. The second imaging system determines the changes of the height of the sample by means of the differences of the positions at which the second receiving device acquires the second images, and performs automatic compensation according to the changes of the height of the sample, so as to ensure that the sample is always at a focused position. The embodiment can perform light path adjustment of the N separate laser light paths respectively by means of the second imaging system, so as to realize that the N separate laser light paths have the same penetration depth. Therefore no further light path adjustment is required when switching between different light sources, thus saving the time consuming for light path adjustment. ## Claims ### Claim 1 A total internal reflection fluorescence imaging system, comprising: a first imaging system, wherein the first imaging system comprises N separate laser light paths, an objective lens, a first receiving device, N first light sources, a reflecting mirror, and N first dichroic mirrors, N is an integer greater than one, each laser light path of the N separate laser light paths is configured to generate light to enter the objective lens, the light passing through the objective lens illuminates a sample to excite the sample to emit fluorescence, the fluorescence passes through the objective lens and enters the first receiving device to form a first image, wherein each of the N first light sources corresponds to one of the N first dichroic mirrors, wherein the N separate laser light paths are composed by the N first light sources, the reflecting mirror, and the N first dichroic mirrors, and wherein light generated in each of the laser light paths is emitted from the first light source, the light emitted from the first light source is reflected by the reflecting mirror to enter the first dichroic mirror, the first dichroic mirror reflects the light into the objective lens; and a second imaging system, wherein the second imaging system comprises a light emitting source, the objective lens, and a second receiving device, the light emitting source is configured to emit light to enter the objective lens, the light passing through the objective lens illuminates the sample and is reflected, the reflected light passes through the objective lens and enters the second receiving device to form a second image. ### Claim 2 (depends on 1) An imaging system according to claim 1, wherein the second imaging system further comprises a compensation lens and a second dichroic mirror, the light emitting source is a second light source, light emitted from the second light source enters the second dichroic mirror through the compensation lens, the second dichroic mirror reflects the light into the objective lens; wherein the objective lens, the first dichroic mirror, the second dichroic mirror and the first receiving device are configured along a single linear optical axis. ### Claim 3 (depends on 2) An imaging system according to claim 2, wherein the first light source comprises a laser, a diverging lens, a collimating lens, a first bandpass filter, a field stop, and a third dichroic mirror, wherein all of optical members constituting the first light source are configured along a single linear optical axis. ### Claim 4 (depends on 3) An imaging system according to claim 3, wherein the lasers comprised in the N first light sources are lasers of different operating wavelengths. ### Claim 5 (depends on 4) An imaging system according to claim 4, wherein optical members comprised in the N first light sources are corresponding to lasers of different operating wavelengths comprised in the N first light sources. ### Claim 6 (depends on 1) An imaging system according to claim 1, wherein the first receiving device comprises a tube lens, a second bandpass filter, and an electron multiplying camera, wherein all of optical members constituting the first receiving device are configured along a single linear optical axis. ### Claim 7 (depends on 6) An imaging system according to claim 6, wherein the first light source comprises a third dichroic mirror and wherein the first dichroic mirror and the third dichroic mirror form an angle of 45 degree to the horizontal direction. ### Claim 8 (depends on 2) An imaging system according to claim 2, wherein the second dichroic mirror forms an angle of 135 degree to the horizontal direction. ### Claim 9 (depends on 1) An imaging system according to claim 1, wherein the objective lens is a total internal reflection objective. ### Claim 10 (depends on 1) An imaging system according to claim 1, wherein the light emitting source is an LED light source. ### Claim 11 (depends on 1) A sequencing device, comprising an imaging system according to claim 1. ### Claim 12 (depends on 2) A sequencing device, comprising an imaging system according to claim 2.