System and method for high numeric aperture imaging systems

• US 7,719,598 B2
• Filed: 01/24/2006
• Issued: 05/18/2010
• Est. Priority Date: 10/12/2000
• Status: Expired due to Fees

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Amnis Corporation (DiaSorin S.p.A.)
• Inventors: Basiji, David A., Perry, David J., Ortyn, William E.
• Primary Examiner(s): HO, TUAN V
• Application Number:
  US11/338,477
• Publication Number:
  US 20060119731A1
• Time in Patent Office: 1,575 Days
• Field of Search: 348335-339, 348/345, 396/111
• US Class Current: 348/335
• 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. An imaging system comprising:

a first subsystem configured to obtain a first image using light propagating along a first optical axis just before the light used to obtain the first image intersects a first focal plane, the first focal plane lying generally perpendicular to the first optical axis; and

a second subsystem configured to obtain a second image, simultaneously with the obtaining of the first image by the first subsystem, using light in the second subsystem that is also propagating along the first optical axis just before the light used to obtain the second image intersects a second focal plane, the second focal plane lying generally parallel to the first focal plane.

2. The imaging system of claim 1, wherein a depth of field of the first subsystem overlaps a depth of field of the second subsystem.

3. The imaging system of claim 1, comprising:

an element configured to collect light from a target object;

an element configured to split the collected light into at least two optical paths; and

wherein the first and second subsystems are configured to obtain the respective first and second images by focusing the light of a respective one of the at least two optical paths according to a position of the respective focal plane.

4. The imaging system of claim 1, 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.

5. The imaging system of claim 1, further comprising:

third and fourth subsystems configured to obtain third and fourth images, respectively, 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.

6. The imaging system of claim 1, wherein the first subsystem is configured to obtain the first image by collection of at least one of reflected light, transmitted light, scattered light, and emitted light from a target object.

7. The imaging system of claim 6, wherein the second subsystem is configured to obtain the second image by collection of at least one of reflected light, transmitted light, scattered light, and emitted light from the target object.

8. The imaging system of claim 6, wherein emitted light comprises fluorescent light.

9. The imaging system of claim 1, wherein the first and second subsystems are each configured to obtain an image in at least one of a plurality of segments of spectral bands.

10. The imaging system of claim 1, further comprising:

a third subsystem configured to obtain a third image, simultaneously with the obtaining of the first image by the first subsystem, using light propagating along a second optical axis just before the light used to obtain the third image intersects a third focal plane, the second optical axis lying substantially perpendicular to the first optical axis, and the third focal plane lying substantially perpendicular to the first focal plane.

11. The imaging system of claim 10, 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 a target object and an additional target object, as detected by the third subsystem.

12. The imaging system of claim 10, further comprising:

a fourth subsystem configured to obtain a fourth image, simultaneously with the obtaining of the first image by the first subsystem, using light propagating along the second optical axis and intersecting a fourth focal plane, parallel to the third focal plane.

13. The imaging system of claim 1, further comprising:

an image detector, the first and second subsystems being controllable to focus the respective first and second images on the image detector;

a motor driver configured to change the focus of the first and second subsystems in response to focus error signals; and

a host computer configured to control image acquisition by the first and second subsystems by transmitting focus error signals to the motor driver.

14. The imaging system of claim 13, wherein the host computer is configured to compare a frequency content of the first and second images, and to transmit a focus error signal to the motor driver if the frequency content of the first and second images is not balanced.

15. A method of obtaining an image, comprising the steps of:

obtaining a first image using light propagating along an optical axis just before the light used for obtaining the first image intersects a first focal plane that is generally perpendicular to the optical axis; and

obtaining, simultaneously with obtaining the first image, a second image using light also propagating along the optical axis just before the light used to obtain the second image intersects a second focal plane, different from, and generally parallel to, the first focal plane.

16. The method of claim 15, further comprising the steps of:

collecting light from a target object;

splitting the light that was collected into a plurality of optical paths;

focusing a first one of the plurality of optical paths on the first focal plane to obtain the first image; and

focusing a second one of the plurality of optical paths on the second focal plane to obtain the second image.

17. The method of claim 15, wherein:

the step of obtaining the first image comprises the step of obtaining the first image at a first depth of field; and

the step of obtaining the second image comprises the step of obtaining the second image at a second depth of field, the first and second depths of field overlapping.

18. The method of claim 15, comprising obtaining, simultaneously with obtaining the first and second images, a third image of the target object along the optical axis at a third focal plane, different from, and parallel with, the first and second focal planes.

19. The method of claim 15, further comprising receiving the first and second obtained images at axially disparate positions.

20. The method of claim 15, further comprising the steps of:

comparing elements of the first and second images; and

adjusting positions of the first and second focal planes if the elements of the first and second images are not balanced.

21. The method of claim 20, wherein the step of adjusting the positions of the first and second focal planes comprises the step of producing a focus error signal determined from a ratio of an imbalance of the elements of the first and second images.

22. The method of claim 20, wherein the elements of the first and second images are spectral segments of the respective first and second images, and wherein the step of comparing the elements comprises the step of comparing a frequency content of the respective first and second images.

23. An imaging system comprising:

a first subsystem configured to obtain a first image at a first depth of field using light propagating along a first optical axis so as to intersect a first focal plane, the first focal plane lying generally perpendicular to the first optical axis; and

a second subsystem configured to obtain a second image at a second depth of field, simultaneously with the obtaining of the first image by the first subsystem, using light in the second subsystem that also propagates along the first optical axis to intersect a second focal plane, the second focal plane lying generally parallel to the first focal plane, wherein the first depth of field and the second depth of field are different, but overlap.

24. A method of obtaining an image, comprising the steps of:

obtaining a first image at a first depth of field, using light propagating along an optical axis, intersecting a first focal plane; and

simultaneously with the step of obtaining the first image, obtaining a second image at a second depth of field, using light also propagating along the optical axis, intersecting a second focal plane that is different from and generally parallel to the first focal plane, wherein the first depth of field and the second depth of field are different, but overlap.