Miniaturized microscope for phase contrast and multicolor fluorescence imaging

• US 10,634,890 B1
• Filed: 10/26/2018
• Issued: 04/28/2020
• Est. Priority Date: 10/26/2018
• Status: Active Grant

• Current Assignee: General Electric Company
• Original Assignee: General Electric Company
• Inventors: Kasten, Ansas Matthias, Challener, William Albert, Karp, Jason Harris, Mandal, Sudeep
• Primary Examiner(s): Taningco, Marcus H
• Application Number:
  US16/171,707
• Publication Number:
  US 20200132975A1
• Time in Patent Office: 550 Days
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6478 Special lenses
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/0221 Portable; cableless; compact; hand-held
• CPC Class Codes: G02B 21/0008 Microscopes having a simple construction, e.g. portable microscopes
• CPC Class Codes: G02B 21/0032 Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06
• CPC Class Codes: G02B 21/0044 moving apertures, e.g. Nipkow disks, rotating lens arrays
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/086 for transillumination only
• CPC Class Codes: G02B 21/18 Arrangements with more than one light path, e.g. for comparing two specimens
• CPC Class Codes: G02B 21/241 Devices for focusing focusing in general G02B7/28
• CPC Class Codes: G02B 21/34 Microscope slides, e.g. mounting specimens on microscope slides
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
• CPC Class Codes: G02B 27/32 Fiducial marks and measuring scales within the optical system

Abstract

A microscope is provided. The microscope includes an illumination source configured to provide illumination beams to image a portion of a biological sample. The microscope also includes an optical unit configured to enable both phase contrast imaging and multicolor fluorescence imaging of the portion of the biological sample utilizing parallel point scanning. The microscope further includes a detector configured to simultaneously acquire multiple point images at different locations of the portion of the biological sample.

Claims

1. A microscope, comprising:

an illumination source configured to provide illumination beams to image a portion of a biological sample;

an optical unit configured to enable both phase contrast imaging and multicolor fluorescence imaging of the portion of the biological sample utilizing parallel point scanning; and

a detector configured to simultaneously acquire a plurality of point images at different locations of the portion of the biological sample.

2. The microscope of claim 1, wherein the microscope is portable and configured to be handheld.

3. The microscope of claim 1, wherein the optical unit has an overall thickness of less than 5 millimeters.

4. The microscope of claim 1, wherein the optical unit comprises a first microlens array and an edge long-pass filter disposed on a backside of the first microlens array, wherein the edge long-pass filter is configured to form a respective condenser annulus associated with a respective lens element of the first microlens array, and each respective condenser annulus is configured to block a center portion of an illumination beam at a first wavelength on the respective lens element and to enable the illumination beam at the first wavelength incident upon the respective lens element through the respective condenser annulus to be focused on the portion of biological sample, and wherein the edge long-pass filter is transparent for illumination beams at one or more wavelengths different from the first wavelength.

5. The microscope of claim 4, wherein the optical unit comprises a second microlens array focally aligned with the first microlens array, wherein the second microlens array is configured to collimate a diffracted and phase altered transmitted beam at the first wavelength from the biological sample, and the second microlens array is configured to collect fluorescent light emitted by the biological sample at the one or more wavelengths.

6. The microscope of claim 5, comprising a microarray phase plate configured to retard by 90 degrees the collimated phase altered transmitted beam and to enable undiffracted light at the first wavelength that passed through biological sample to pass through.

7. The microscope of claim 6, comprising an absorption filter and an interference notch filter located downstream of the first and second microlens array and the microarray phase plate, wherein the absorption filter and the interference notch filter are transparent to light at the first wavelength and filter the emitted fluorescent light at the one or more wavelengths.

8. The microscope of claim 7, comprising a third microlens array and a fourth microlens array focally aligned with the first and second microlens array and located downstream of the absorption filter and the interference notch filter, and a pinhole array disposed between the third and fourth microlens array, and wherein the third microlens array, the pinhole array, and the fourth microlens array are configured to spatially filter the phase altered transmitted beam and the undiffracted light at the first wavelength and to spatially filter the emitted fluorescent light at the one or more wavelengths for detection by the detector.

9. The microscope of claim 7, comprising a dichroic mirror disposed between the second microlens array and the microarray phase plate that is reflective at the second and third wavelengths.

10. The microscope of claim 1, comprising a piezo-driven scanner configured to move the biological sample in multiple directions, wherein the piezo-driven scanner is configured to step the biological sample one lens element pitch in both a first direction and a second direction.

11. The microscope of claim 1, comprising a processor configured to generate a phase contrast image and a multicolor fluorescence image of the portion of the biological sample based on the plurality of point images at different locations.

12. The microscope of claim 1, wherein the microscope is configured to be utilized in a trans-illumination configuration, a total internal reflection-illumination configuration, and an epi-illumination configuration.

13. The microscope of claim 1, comprising a waveguide structure formed by a pair of glass slides surrounding a high refractive index fluid, and a prism configured to provide the illumination beams at the one or more wavelengths to the waveguide structure for illuminating the biological sample.

14. A microscope, comprising:

an optical unit configured to enable both phase contrast imaging and multicolor fluorescence imaging of a portion of a biological sample utilizing parallel point scanning, wherein the optical unit comprises;

a first microlens array;

an edge long-pass filter disposed on a backside of the first microlens array, wherein the edge long-pass filter is configured to form a respective condenser annulus associated with a respective lens element of the first microlens array, wherein the edge long-pass filter is configured to enable a portion of a beam at a first wavelength to pass through to the first microlens array, and the edge-long filter is transparent to beams at a second and a third wavelength different from the first wavelength;

a second microlens array, wherein the second microlens array is configured to collimate a diffracted transmitted beam at the first wavelength from the biological sample, and the second microlens array is configured to collect fluorescent beams emitted by the biological sample at the second wavelength and the third wavelength;

an absorption filter and an interference notch filter located downstream of the first and second microlens array, wherein the absorption filter and the interference notch filter are configured to be transparent to the beam at the first wavelength and block excitation beams of different wavelengths while only allowing emitted fluorescent beams at the second and third wavelengths to pass; and

a third microlens array and a fourth microlens array located downstream of the absorption filter and the interference notch filter, and a pinhole array disposed between the third and fourth microlens array, and wherein the third microlens array, the pinhole array, and the fourth microlens array are configured to spatially filter the transmitted beam at the first wavelength and the undiffracted beam at the first wavelength that has passed through the biological sample and to spatially filter the emitted fluorescent beams at the second and third wavelengths;

wherein the first, second, third, and fourth microlens arrays are focally aligned.

15. The microscope of claim 14, wherein each respective condenser annulus is configured to block a center portion of the beam at the first wavelength on the respective lens element and to enable the beam at the first wavelength incident upon the respective lens element through the respective condenser annulus to be focused on the portion of biological sample.

16. The microscope of claim 14, wherein the optical unit has an overall thickness of less than 5 millimeters.

17. The microscope of claim 14, wherein the optical unit comprises a microarray phase plate configured to retard by 90 degrees the collimated phase altered transmitted beam and to enable the undiffracted beam at the first wavelength that passed through biological sample to pass through.

18. A method of manufacturing an optical unit for a microscope configured to enable both phase contrast imaging and multicolor fluorescence imaging of a portion of a biological sample utilizing parallel point scanning, comprising:

providing a first microlens array;

disposing an edge long-pass filter on a backside of the first microlens array, wherein the edge long-pass filter is configured to form a respective condenser annulus associated with a respective lens element of the first microlens array, wherein the edge long-pass filter is configured to enable a portion of a beam at a first wavelength to pass through to the first microlens array, and the edge-long filter is transparent to beams at a second and a third wavelength different from the first wavelength;

optically aligning a second microlens array with the first microlens array, wherein the second microlens array is configured to collimate a diffracted transmitted beam at the first wavelength from the biological sample, and the second microlens array is configured to collect fluorescent beams emitted by the biological sample at the second wavelength and the third wavelength;

disposing an absorption filter and an interference notch filter downstream of the first and second microlens array, wherein the absorption filter and the interference notch filter are configured to be transparent to the beam at the first wavelength and block excitation beams of different wavelengths while only allowing emitted fluorescent beams at the second and third wavelengths to pass; and

optically aligning a third microlens array, a fourth microlens array, and a pinhole array disposed between the third and fourth microlens array with the first and second microlens arrays downstream of the absorption filter and the interference notch filter, wherein the third microlens array, the pinhole array, and the fourth microlens array are configured to spatially filter the transmitted beam at the first wavelength and the undiffracted beam at the first wavelength that has passed through the biological sample and to spatially filter the emitted fluorescent beams at the second and third wavelengths.

19. The method of claim 18, comprising disposing a microarray phase plate between the second microlens array and the absorption filter, wherein the microarray phase plate is configured to retard by 90 degrees the collimated phase altered transmitted beam and to enable the undiffracted beam at the first wavelength that passed through biological sample to pass through.

20. The method of claim 18, wherein the optical unit has an overall thickness of less than 5 millimeters.