Techniques for scanned illumination

• US 9,488,823 B2
• Filed: 06/03/2013
• Issued: 11/08/2016
• Est. Priority Date: 06/07/2012
• Status: Active Grant

• Current Assignee: Complete Genomics Incorporated (BGI Genomics Co., Ltd.)
• Original Assignee: Complete Genomics Incorporated (BGI Genomics Co., Ltd.)
• Inventors: Staker, Bryan P., Uhrich, Craig E.
• Primary Examiner(s): Anyikire, Chikaodili E
• Application Number:
  US13/908,964
• Publication Number:
  US 20140152793A1
• Time in Patent Office: 1,254 Days
• US Class Current: 1/1
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G02B 21/361 Optical details, e.g. image relay to the camera or image sensor G02B21/364 takes precedence; illumination details G02B21/06 and subgroups
• CPC Class Codes: G02B 21/365 Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence
• CPC Class Codes: G06T 1/0007 Image acquisition
• CPC Class Codes: G06T 2207/10056 Microscopic image
• CPC Class Codes: G06T 2207/10064 Fluorescence image
• CPC Class Codes: G06T 2207/10152 Varying illumination
• CPC Class Codes: G06T 2207/30004 Biomedical image processing
• CPC Class Codes: G06T 2207/30072 Microarray; Biochip, DNA array; Well plate
• CPC Class Codes: H04N 23/56 provided with illuminating means
• CPC Class Codes: H04N 25/441 by reading contiguous pixels from selected rows or columns of the array, e.g. interlaced scanning
• CPC Class Codes: H04N 25/531 by controlling rolling shutters in CMOS SSIS
• CPC Class Codes: H04N 25/75 Circuitry for providing, modifying or processing image signals from the pixel array
• CPC Class Codes: H04N 25/767 Horizontal readout lines, multiplexers or registers
• CPC Class Codes: H04N 25/78 Readout circuits for addressed sensors, e.g. output amplifiers or A/D converters
• CPC Class Codes: H04N 3/08 having a moving reflector
• CPC Class Codes: H04N 7/18 Closed-circuit television [CCTV] systems, i.e. systems in which the video signal is not broadcast

Abstract

Imaging systems are provided for high speed, high resolution imaging of biochemical materials. In an example embodiment, an imaging system comprises an objective lens component, a line generator, a digital camera, a positioning stage, and a scan mirror. The line generator generates a line of light that is scanned across a portion of a substrate that is mounted on the positioning stage. The positioning stage moves the substrate in a particular direction that is substantially normal to an optical axis of the objective lens component. The camera collects an image of the portion of the substrate through the objective lens component. The scan mirror moves in coordination with the positioning stage, while the line of light is being scanned across the portion of the substrate and the substrate is being moved in the particular direction, in order to keep the image still with respect to the camera while the image is being collected by the camera.

Claims

1. A method for operating an imaging system, the method comprising:

collecting a two-dimensional image of a portion of a substrate on a positioning stage via a digital camera in the imaging system by moving the positioning stage in a first path of travel under an objective lens component of the imaging system that is normal to the optical axis of the objective lens component, wherein the image spans multiple rows of pixels of a sensor element in the camera;

while the camera is collecting the image of the portion of the substrate, performing steps comprising;

scanning a line of light across the portion of the substrate by changing an angle of a scan mirror so that the image of the portion of the substrate that is acquired by the objective lens component is kept still with respect to the digital camera while the substrate is moving in a continuous motion, thereby exposing first one or more rows of pixels spanned by the image while keeping in the dark second one or more rows of pixels spanned by the image; and

reading out the first one or more rows of pixels while continuing to scan the line of light across the portion of the substrate thereby exposing the second one or more rows of pixels.

2. The method of claim 1 wherein changing the angle of the scan mirror is performed in coordination with scanning the line of light across the portion of the substrate.

3. The method of claim 1, wherein:

changing the angle of the scan mirror includes a first time interval during which the line of light is scanned across the portion of the substrate and a second time interval during which the scan mirror is returned to an initial position; and

the method further comprises ceasing to scan the line of light across the portion of the substrate during the second time interval.

4. The method of Claim 1, wherein reading out the first one or more rows of pixels is performed while the substrate is being moved in a direction that is normal to the optical axis of the objective lens component.

5. The method of claim 1, wherein scanning the line of light across the portion of the substrate comprises changing an angle of an illumination mirror to reflect light from a line generator onto the portion of the substrate.

6. The method of claim 5, wherein changing the angle of the illumination mirror is effected by a servo mechanism configured to tilt the illumination mirror.

7. The method of claim 5, further comprising:

moving the substrate under an objective lens component of the imaging system;

changing an angle of a scan mirror so that the image of the portion of the substrate that is acquired by the objective lens component is kept still with respect to the camera while the substrate is moving;

wherein changing the angle of the scan mirror is performed in coordination with changing the angle of the illumination mirror.

8. The method of claim 1, further comprising keeping the first one or more rows of pixels dark between the steps of exposing the first one or more rows of pixels and reading out the first one or more rows of pixels.

9. The method of claim 1, wherein the camera comprises a split-readout sensor having at least a first section and a second section, and the steps of scanning the line of light and reading out are performed independently in each section of the split-readout sensor.

10. The method of claim 9, wherein the steps of scanning and reading out in the first section are performed in parallel to the steps of scanning and reading out in the second section.

11. The method of claim 9, wherein the steps of scanning and reading out in the first section are performed in anti-parallel to the steps of scanning and reading out in the second section.

12. The method of claim 1, wherein the line of light has a wavelength appropriate for fluorescence excitation while pixels in the camera are exposed by light having a wavelength corresponding to fluorescence emission.

13. The method of claim 1, further comprising:

while the camera is collecting the image of the portion of the substrate, reading out the second one or more rows of pixels while continuing to scan the line of light across the substrate thereby exposing third one or more rows of pixels spanned by the image, wherein the third one or more rows of pixels are different than the second one or more rows of pixels.

14. The method of claim 13, wherein the third one or more rows of pixels are different than the first one or more rows of pixels.

15. The method of claim 1, wherein the camera is one of a CMOS camera and a non-CMOS camera operating in full-frame mode.

16. A method for operating an imaging system, the method comprising:

collecting an image of a portion of a substrate via a digital camera in the imaging system, wherein the image spans multiple rows of pixels of a sensor element in the camera;

while the camera is collecting the image of the portion of the substrate, performing steps comprising;

scanning a line of light across the portion of the substrate thereby exposing first one or more rows of pixels spanned by the image while keeping in the dark second one or more rows of pixels spanned by the image;

moving the substrate under an objective lens component of the imaging system;

changing an angle of a scan mirror so that the image of the portion of the substrate that is acquired by the objective lens component is kept still with respect to the camera while the substrate is moving; and

reading out the first one or more rows of pixels while continuing to scan the line of light across the portion of the substrate, thereby exposing the second one or more rows of pixels;

wherein changing the angle of the scan mirror includes a first time interval during which the line of light is scanned across the portion of the substrate and a second time interval during which the scan mirror is returned to an initial position;

further including ceasing to scan the line of light across the portion of the substrate during the second time interval; and

wherein the first time interval is greater than the second time interval.

17. The method of claim 16, wherein changing the angle of the scan mirror is performed in coordination with scanning the line of light across the portion of the substrate.

18. The method of claim 16, wherein reading out the first one or more rows of pixels is performed while the substrate is being moved in a direction that is normal to the optical axis of the objective lens component.

19. The method of claim 16, wherein scanning the line of light across the portion of the substrate comprises changing an angle of an illumination mirror to reflect light from a line generator onto the portion of the substrate.

20. The method of claim 16, further comprising keeping the first one or more rows of pixels dark between the steps of exposing the first one or more rows of pixels and reading out the first one or more rows of pixels.

21. The method of claim 16, wherein the camera comprises a split-readout sensor having at least a first section and a second section, and the steps of scanning the line of light and reading out are performed independently in each section of the split-readout sensor.

22. The method of claim 16, wherein the line of light has a wavelength appropriate for fluorescence excitation while pixels in the camera are exposed by light having a wavelength corresponding to fluorescence emission.

23. The method of claim 16, wherein the camera is one of a CMOS camera and a non-CMOS camera operating in full-frame mode.

24. An imaging system comprising:

an objective lens component;

a line generator that is configured to generate a line of light that is scanned across a portion of a substrate;

a positioning stage that is configured to move the substrate in a first direction path of travel that is substantially normal to an optical axis of the objective lens component;

a digital camera that is configured to acquire a two-dimensional image of the portion of the substrate through the objective lens component; and

a scan mirror that is configured to move in coordination with the positioning stage, while the line of light is being scanned across the portion of the substrate and the substrate is being moved in the first direction path of travel in a continuous motion, in order to maintain the image still with respect to the camera while the image is being acquired by the camera; and

an illumination mirror and a computer logic, wherein the image spans multiple rows of pixels in the camera, and wherein while the camera is acquiring the image of the portion of the substrate;

the illumination mirror is configured and operative to scan the line of light across the portion of the substrate to expose first one or more rows of pixels spanned by the image while keeping in the dark second one or more rows of pixels spanned by the image; and

the computer logic is configured and operative to read out the first one or more rows of pixels while the illumination mirror continues to scan the line of light across the portion of the substrate to expose the second one or more rows of pixels.

25. The imaging system of claim 24, further comprising an illumination mirror configured to scan the line of light from the line generator onto the portion of the substrate, wherein the scan mirror is configured to move in coordination with the illumination mirror while the line of light is being scanned across the portion of the substrate and the substrate is being moved in the particular direction.

26. The imaging system of claim 25, further comprising:

a first servo mechanism configured to change an angle of the illumination mirror to scan the line of light across the portion of the substrate; and

a second servo mechanism configured to change angle of the scan mirror in coordination with changes to the angle of the illumination mirror.

27. The imaging system of claim 26, wherein:

the second servo mechanism is configured to change angle of the scan mirror during a first time interval in which the line of light is scanned across the portion of the substrate;

the second servo mechanism is configured to return the scan mirror to an initial position during a second time interval in which the line of light is not scanned across the portion of the substrate, wherein the second time interval is shorter than the first time interval.

28. The imaging system of claim 24, further comprising a dichroic mirror that is configured at least to:

(a) reflect the line of light from the illumination mirror onto the scan mirror in order to illuminate the portion of the substrate; and

(b) pass to the camera light acquired by the objective lens component and is reflected by the scan mirror.

29. The imaging system of claim 24, wherein the camera comprises a split-readout sensor having at least two sections that can be exposed and read out independently of each other.

30. The imaging system of claim 24, wherein the camera is one of a CMOS camera and a non-CMOS camera operating in full-frame mode.