# US 8,542,274 B2 · Wide field microscopic imaging system and method > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,542,274 B2 - Document ID: US8542274B2 - Status: Granted - Kind: B2 - Application number: 12/906,411 - Filed: 2010-10-18 - Published: 2012-04-19 - Granted: 2013-09-24 - Earliest claimed: 2010-10-18 - Assignees: Olympus America Inc. - Applicants: Yoshihiro Kawano; Lorne D. Davies - Inventors: Yoshihiro Kawano; Lorne D. Davies - Web page: https://patentgrep.com/patent/US8542274B2 ## CPC - G02B 21/365: {Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take precedence)} - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS ## Abstract A wide field microscopic image acquisition apparatus and method are disclosed. The apparatus is configured to acquire images of a specimen on a microscope slide and includes first and second illuminators each having unique illumination characteristics. The apparatus includes a microscope imaging system with an imaging device, an objective lens and a stage configured to digitally acquire a plurality of images of the specimen using the first and second illuminators. A controller is configured to automatically control the microscope imaging system and acquire the plurality of images of the specimen using the first and second illuminators. The first and second illuminators can be bright field, dark field or fluorescent illuminators. ## Claims ### Claim 1 A wide field microscopic image acquisition apparatus for acquiring images of a specimen on a microscope slide, the apparatus comprising: first and second illuminators each having unique illumination characteristics; a microscope imaging system including an imaging device, an objective lens and a stage configured to digitally acquire a plurality of images of the specimen using the first and second illuminators; and a controller configured to automatically control the microscope imaging system and scan the specimen on the microscope slide to acquire the plurality of images from a scan area of the specimen using the first and second illuminators; wherein the microscope slide defines a first side of the microscope slide facing to the objective lens and a second side of the slide perpendicular to the first side; the first illuminator comprises a dark field illuminator configured to direct light into the second side of the microscope slide; the second illuminator comprises a fluorescent illuminator; and the controller is configured to acquire a first set of images of the scan area using the dark field illuminator, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using the fluorescent illuminator based on the stored dark field focus map. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein the controller is configured to acquire images of the specimen using the first illuminator and the second illuminator prior to moving the specimen in an X-Y direction with respect to the objective lens. ### Claim 3 (depends on 1) The apparatus of claim 1, further comprising a condenser, wherein the imaging device and objective lens have a light path and the controller is configured to adjust the condenser to minimize light transmitted along the light path. ### Claim 4 (depends on 3) The apparatus of claim 3, wherein the condenser has a pivot and the controller is configured to move the condenser via the pivot to minimize light transmitted along the light path. ### Claim 5 (depends on 1) The apparatus of claim 1, wherein the controller is configured to digitally acquire a plurality of Z images of the specimen. ### Claim 6 (depends on 1) The apparatus of claim 1, wherein the controller is configured to acquire a plurality of images of the specimen at multiple magnifications. ### Claim 7 (depends on 1) The apparatus of claim 1, wherein the controller is configured to save a data structure of images of the specimen. ### Claim 8 (depends on 1) The apparatus of claim 1, wherein the first illuminator comprises a first light source having a first spectral output and a second light source having a second spectral output, the first and second light sources directing light into the second side of the microscope slide. ### Claim 9 (depends on 1) The apparatus of claim 1, wherein the first and second light sources are configured to direct light into the same side of the microscope slide. ### Claim 10 (depends on 1) The apparatus of claim 1, wherein the first illuminator comprises a light guide configured to collimate light originating from the dark field illuminator. ### Claim 11 (depends on 10) The apparatus of claim 10, wherein the first illuminator comprises a plurality of individual light sources arranged in array and the light guide is formed with upper and lower portions, configured to collimate light originating from the individual light sources. ### Claim 12 (depends on 1) The apparatus of claim 1, wherein the first illuminator comprises a plurality of individual light sources arranged in array, wherein the individual light sources are configured to be selectively energized. ### Claim 13 (depends on 1) The apparatus of claim 1, wherein the first illuminator comprises a plurality mounting units configured to hold a plurality of unpackaged individual light sources. ### Claim 14 A method of acquiring wide field microscopic images of a specimen on a microscope slide, the method comprising: providing first and second illuminators each having unique illumination characteristics; providing a microscope imaging system including an imaging device, an objective lens and a stage configured to digitally acquire a plurality of images from a scan area of the specimen using the first and second illuminators; and providing a controller configured to automatically control the microscope imaging system and scan the specimen on the microscope slide to acquire the plurality of images from the scan area using the first and second illuminators; wherein the microscope slide defines a first side of the microscope slide facing to the objective lens and a second side of the slide perpendicular to the first side; the first illuminator comprises a dark field illuminator that directs light into the second side of the microscope slide; the second illuminator comprises a fluorescent illuminator; and the controller is configured to acquire a first set of images of the scan area using the dark field illuminator, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using the fluorescent illuminator based on the stored dark field focus map. ### Claim 15 (depends on 14) The method of claim 14, wherein the controller is configured to acquire a plurality of X-Y images of the specimen using the first illuminator and the second illuminator prior to switching to the second illuminator. ### Claim 16 (depends on 14) The method of claim 14, wherein the controller is configured to acquire images of the specimen using the first illuminator prior to moving the specimen in an X-Y direction with respect to the objective lens. ### Claim 17 (depends on 14) The method of claim 14, further comprising providing a condenser, wherein the imaging device and objective lens have a light path and the controller is configured to adjust the condenser to minimize light transmitted along the light path. ### Claim 18 (depends on 17) The method of claim 17, wherein the condenser has a pivot and the controller is configured to move the condenser via the pivot to minimize light transmitted along the light path. ### Claim 19 (depends on 14) The method of claim 14, wherein the controller is configured to acquire a plurality of images of the specimen at multiple magnifications. ### Claim 20 (depends on 14) The method of claim 14, wherein the controller is configured to save a data structure of images of the specimen. ### Claim 21 (depends on 14) The method of claim 14, wherein the first illuminator comprises a first light source having a first spectral output and a second light source having a second spectral output, the first and second light sources directing light into the second side of the microscope slide. ### Claim 22 (depends on 21) The method of claim 21, wherein the first and second light sources are configured to direct light into the same side of the microscope slide. ### Claim 23 (depends on 14) The method of claim 14, wherein the first illuminator comprises a light guide configured to collimate light originating from the dark field illuminator. ### Claim 24 (depends on 23) The method of claim 23, wherein the first illuminator comprises a plurality of individual light sources arranged in array and the light guide is formed with upper and lower portions, configured to collimate light originating from the individual light sources. ### Claim 25 (depends on 14) The method of claim 14, wherein the first illuminator comprises a plurality of individual light sources arranged in array, wherein the individual light sources are configured to be selectively energized. ### Claim 26 (depends on 14) The method of claim 14, wherein the first illuminator comprises a plurality mounting units configured to hold a plurality of unpackaged individual light sources. ### Claim 27 A wide field microscopic image acquisition apparatus for acquiring images of a specimen on a microscope slide, the apparatus comprising: an illuminator configured to direct light into a side of the microscope slide and having a dark field light source with a first spectral output and a second light source with a second spectral output; a microscope imaging system including an imaging device and an objective lens and a stage configured to digitally acquire a plurality of images of the specimen; and a controller configured to automatically control the microscope imaging system and acquire the plurality of images of the specimen the controller being configured to acquire a first set of images of the scan area using the dark field light source, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using the second light source based on the stored dark field focus map; wherein the microscope slide defines a first side of the microscope slide facing to the objective lens and a second side of the slide perpendicular to the first side; and the dark field and second light sources direct light into the second side of the microscope slide, the controller being configured to acquire a first set of images of the scan area using the dark field light source, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using the second light source based on the stored dark field focus map; wherein the microscope slide defines a first side of the microscope slide facing to the objective lens and a second side of the slide perpendicular to the first side; and the dark field and second light sources direct light into the second side of the microscope slide. ### Claim 28 A wide field microscopic image acquisition apparatus for acquiring images of a specimen on a microscope slide, the apparatus comprising: an illuminator with a plurality of light sources configured to direct light into a side of the microscope slide; a light guide configured to collimate light originating from the plurality of light sources; a microscope imaging system including an imaging device and an objective lens and a stage configured to digitally acquire a plurality of images of the specimen; and a controller configured to automatically control the microscope imaging system and acquire the plurality of images of the specimen a controller configured to automatically control the microscope imaging system and acquire the plurality of images of the specimen, the controller being configured to acquire a first set of images of the scan area using a dark field light source, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using a second light source based on the stored dark field focus map, the controller being configured to acquire a first set of images of the scan area using a dark field light source, generate a dark field focus map defining a focal depth for each of the images within the first set of images, store the dark field focus map and acquire a second set images of the scan area using a second light source based on the stored dark field focus map; wherein the microscope slide defines a first side of the microscope slide facing to the objective lens and a second side of the slide perpendicular to the first side; and a plurality of light sources direct light into the second side of the microscope slide.