System and method for high numeric aperture imaging systems

• US 20060119731A1
• Filed: 01/24/2006
• Published: 06/08/2006
• Est. Priority Date: 10/12/2000
• Status: Active Grant

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Amnis Corporation (DiaSorin S.p.A.)
• Inventors: Basiji, David A., Perry, David J., Ortyn, William E.
• Granted Patent: US 7,719,598 B2
• Time in Patent Office: Days
• US Class Current: 348/345
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: C07K 1/047 Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2563/179 the label being a nucleic acid
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/0294 Particle shape
• CPC Class Codes: G01N 2015/1443 Auxiliary imaging
• CPC Class Codes: G01N 2015/1452 Adjustment of focus; Alignment
• CPC Class Codes: G01N 2015/1472 with colour
• CPC Class Codes: G01N 2015/1479 Using diffuse illumination or excitation
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/052 Tubular type; cavity type; multireflective
• CPC Class Codes: G01N 2021/058 Flat flow cell
• CPC Class Codes: G01N 21/031 Multipass arrangements
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G02B 21/00 Microscopes
• CPC Class Codes: G02B 27/0012 : Optical design, e.g. procedures, algorithms, optimisation routines
• CPC Class Codes: G02B 27/1086 : operating by diffraction only
• CPC Class Codes: G02B 27/126 : The splitting element being a prism or prismatic array, including systems based on total internal reflection
• CPC Class Codes: G02B 27/144 : using partially transparent surfaces without spectral selectivity G02B27/147 takes precedence
• CPC Class Codes: G02B 27/145 : having sequential partially reflecting surfaces
• CPC Class Codes: G02B 7/28 : Systems for automatic generation of focusing signals
• CPC Class Codes: H04N 23/673 : based on contrast or high frequency components of image signals, e.g. hill climbing method
• CPC Class Codes: H04N 23/959 : by adjusting depth of field during image capture, e.g. maximising or setting range based on scene characteristics

Abstract

A system and method for high numeric aperture imaging systems includes a splitter, a defocusing system, and a combiner. The splitter reflects a portion of collected light and transmits another portion of the collected light. The defocusing system is configured to modify optical power of either the transmitted portion or reflected portion of the collected light. The combiner is oriented with respect to a mechanical angle. The combiner recombines portions of the transmitted portion and the reflected portion such that the transmitted portion and reflected portion are subsequently transmitted being separated by an optical separation angle based upon the mechanical angle of orientation of the combiner. Various other implementations are used to maintain focus with regards to the imaging systems involved.

Claims

1. For imaging a target object, an imaging system comprising:

a first imaging sub-system comprising a light collection system positioned with respect to the target object in a first orientation, the first imaging sub-system configured to transmit a first image of the target object;

a first detector positioned to receive the first image of the target object at a first location on the first detector when the target object is in a first target location and to receive the first image of the target object at a second location on the first detector when the target object is in a second location different than the first location of the target object;

a second imaging sub-system comprising a light collection system positioned with respect to the target object in a second orientation different than the first orientation, the second imaging sub-system configured to transmit an second image of the target object at a best focus distance from the second imaging sub-system;

a second detector positioned to receive the second image of the target object; and

a processor communicatively linked to the first detector and configured to determine a distance between the first location on the first detector and the second location on the first detector, the processor communicatively linked to the second imaging sub-system and configured to transmit instructions to the second imaging sub-system based upon the determined distance between the first location on the first detector and the second location on the first detector, the second imaging sub-system configured to change the best focus distance from the second imaging sub-system based upon the instructions received from the processor to correct focus.

2. The system of claim 1 wherein the first orientation is along an x-axis and the second orientation is along a y-axis, the x-axis and y-axis being substantially perpendicular to one another.

3. For imaging a target object in a flow stream, an imaging system comprising:

a first imaging sub-system comprising a light collection system positioned with respect to the target object in a first orientation, the first imaging sub-system configured to transmit a first image of the target object;

a first detector positioned to receive the first image of the target object at a first location on the first detector when the target object is in a first target location and to receive the first image of the target object at a second location on the first detector when the target object is in a second location;

a second imaging sub-system comprising a light collection system positioned with respect to the target object in a second orientation, the second imaging sub-system configured to transmit an second image of the target object;

a second detector positioned a distance along an optical axis from the second imaging sub-system to receive light from the second imaging sub-system; and

a processor communicatively linked to the first detector, the processor configured to determine a distance between the first location on the first detector and the second location on the first detector, the processor communicatively linked to the second imaging sub-system, the processor configured to transmit instructions to the second imaging sub-system based upon the determined distance between the first location on the first detector and the second location on the first detector, the second imaging sub-system configured to change the distance that the second detector is positioned along the optical axis from the second imaging sub-system based upon the instructions received from the processor.

4. The system of claim 3 wherein the first imaging sub-system has a magnification and the instructions are based on the determined distance multiplied by the square of the magnification of the second imaging sub-system.

5. A method for maintaining focus in an imaging system comprising:

segmenting objects of interest;

analyzing frequency content of one or more imaged objects in two focus areas associated with received unaltered and defocused light;

if frequency content is not balanced, calculating a focal error signal;

if frequency content is not balanced, determining focal shift; and

adjusting refocusing optics based upon the determined focal shift and the focal error signal.

6. The method of claim 5 wherein the frequency content of the one or more imaged objects in the two focus areas is balanced if the frequency content in the two focus areas is equal.

7. The method of claim 5 wherein the unaltered and defocused light is received on one detector.

8. An imaging system comprising:

a first subsystem configured to obtain a first image of a target object along a first viewing axis in a first focal plane, the first focal plane lying perpendicular to the first optical axis; and

a second subsystem configured to obtain a second image of the target object, simultaneously with the obtaining of the first image by the first subsystem, along the first optical axis in a second focal plane, the second focal plane lying parallel to the first focal plane.

9. The imaging system of claim 8 wherein a depth of field of the first subsystem overlaps a depth of field of the second subsystem.

10. The imaging system of claim 8 wherein the first and second subsystems are configured to divide between them light emanating from the target object into first and second optical paths, respectively, and to obtain the respective first and second images by focusing the light of the respective optical path according to a position of the respective focal plane.

11. The imaging system of claim 8, further comprising an image detector, and wherein the first and second subsystems are configured to focus the respective first and second images on the image detector.

12. The imaging system of claim 8, further comprising:

third and fourth subsystems configured to obtain third and fourth images, respectively, of the target object, simultaneously with the obtaining of the first image by the first subsystem, along the first optical axis in respective third and fourth focal planes, the third and fourth focal planes each lying parallel to the first focal plane.

13. The imaging system of claim 8 wherein the first subsystem is configured obtain the first image by collection of at least one of reflected, transmitted, scattered, and emitted light from the target object.

14. The imaging system of claim 13 wherein the second subsystem is configured obtain the second image by collection of at least one of reflected, transmitted, scattered, and emitted light from the target object.

15. The imaging system of claim 13 wherein emitted light comprises fluorescent light.

16. The imaging system of claim 8 wherein the first and second subsystems are each configured to obtain an image from each of a plurality of segments of spectral bands.

17. The imaging system of claim 8, further comprising:

third and fourth subsystems configured to obtain third and fourth images, respectively, of the target object, simultaneously with the obtaining of the first image by the first subsystem, along a second optical axis in respective third and fourth focal planes, the second optical axis lying substantially perpendicular to the first optical axis, and the third and fourth focal planes each lying substantially perpendicular to the first focal plane.

18. The imaging system of claim 17, further comprising a processor configured to adjust a position of the first focal plane in response to a separation along the first optical axis of the target object and an additional target object, as detected by the third subsystem.