System and method for Hilbert phase imaging

• US 10,256,262 B2
• Filed: 06/10/2014
• Issued: 04/09/2019
• Est. Priority Date: 03/25/2005
• Status: Active Grant

• Current Assignee: Hamamatsu Photonics KK, Massachusetts Institute of Technology
• Original Assignee: Hamamatsu Photonics KK, Massachusetts Institute of Technology
• Inventors: Popescu, Gabriel, Dasari, Ramachandra, Feld, Michael, Ikeda, Takahiro
• Primary Examiner(s): Legasse, Jr., Francis M
• Application Number:
  US14/301,187
• Publication Number:
  US 20140361148A1
• Time in Patent Office: 1,764 Days
• CPC Class Codes: G01N 15/10 Investigating individual particles
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/012 Red blood cells
• CPC Class Codes: G01N 2015/016 White blood cells
• CPC Class Codes: G01N 2015/145 by pattern of light, e.g. fringe pattern
• CPC Class Codes: G01N 2015/1454 using phase shift or interference, e.g. for improving contrast
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 21/453 Holographic interferometry for dimensional measurements G01B9/021 - G01B9/029
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80
• CPC Class Codes: G06V 20/693 Acquisition
• CPC Class Codes: H01L 27/14601 Structural or functional details thereof

Abstract

Hilbert phase microscopy (HPM) as an optical technique for measuring high transverse resolution quantitative phase images associated with optically transparent objects. Due to its single-shot nature, HPM is suitable for investigating rapid phenomena that take place in transparent structures such as biological cells. A preferred embodiment is used for measuring biological systems including measurements on red blood cells, while its ability to quantify dynamic processes on the millisecond scale, for example, can be illustrated with measurements on evaporating micron-size water droplets.

Claims

1. A quantitative phase imaging device comprising:

a light source that generates light transmitted along a first optical path and a second optical path, the light transmitted on the first optical path illuminating a full field of view of a sample;

an imaging device positioned relative to the first optical path and the second optical path such that light from the sample positioned relative to the first optical path is detected by the imaging device and light from the second optical path is tilted at an angle relative to the light from the sample that is incident on the imaging device to provide an interference pattern, the imaging device generating image data of the sample;

a processor that processes the image data with a Hilbert transform to provide a full field quantitative phase image of the sample.

2. The device of claim 1 further comprising a rotating mirror.

3. The device of claim 1 wherein the imaging device collects at least 10 frames per second.

4. The device of claim 1 wherein the processor performs the Hilbert transform on the image data.

5. The device of claim 1 wherein the device comprises an interferometer.

6. The device of claim 1 wherein the device comprises a microscope.

11. The device of claim 1 further comprising performing a Fourier transform on the image data.

12. The device of claim 1 wherein the device comprises an inverted microscope.

13. The device of claim 1 further comprising a lens to receive light from the sample, a combiner to combine light from the lens that is tilted at an angle with light from the second optical path.

14. The device of claim 13 further comprising a lens between the combiner and the imaging device.

7. A method of imaging an object comprising:

splitting light from a light source along a first optical path including an object to be imaged and a second reference optical path;

combining light from the object along the first optical path and the second reference optical path with a combiner, the first optical path being tilted at an angle relative to the second optical path to form a fringe structure at a two-dimensional imaging device having a plurality of pixels;

detecting a two dimensional image of the fringe structure with the two-dimensional imaging device; and

processing the two dimensional image with a data processor configured to apply a Hilbert transform to obtain quantitative phase image data.

8. The method of claim 7 further comprising using a fiber optic device to couple light from the light source to the object.

9. The method of claim 7 wherein the second reference optical path couples a reference field light beam to the imaging device.

10. The method of claim 9 wherein the reference field beam is tilted at the angle with respect to a sample field beam transmitted through, or reflected by, the object.

15. The method of claim 7 further comprising imaging a biological sample.

16. The method of claim 7 further comprising generating a plurality of images in sequence to image a dynamic process.

17. A method of imaging a dynamic biological process comprising:

illuminating a biological material with light from a light source;

detecting light from the biological material with a two dimensional imaging detector having a plurality of pixels to generate a plurality of images, the light from the biological material that is incident upon the imaging detector being combined with reference light that is tilted at an angle relative to the light from the biological material that is incident upon the detector form a fringe pattern; and

processing the images with a data processor programmed to perform a Hilbert transform of each image to generate quantitative phase images of the biological material.

18. The method of claim 17 further comprising coupling the illuminating light to the biological material with a fiber optic device to generate the quantitative phase images.

19. The method of claim 17 wherein the light source and imaging detector comprises a microscope, the microscope displaying images on a display.

20. The method of claim 17 further comprising simultaneously illuminating a full field view of the biological material to generate at least 10 image frames per second of full field images of the biological material.