Method for multi-color fluorescence imaging under single exposure, imaging method and imaging system

Patent metadata

Field	Value
Publication number	US-10823945-B2
Application number	US-201715615180-A
Country	US
Kind code	B2
Filing date	Jun 6, 2017
Priority date	Jan 10, 2017
Publication date	Nov 3, 2020
Grant date	Nov 3, 2020

Abstract

The present disclosure relates to a method for multi-color fluorescence imaging under a single exposure, an imaging method and system. The imaging system includes: a fluorescence microscope, configured to obtain a real image of the sample; a spatial mask, disposed behind the fluorescence microscope, and configured to perform mask modulation on the real image of the sample; a 4f system, disposed behind the spatial mask, in which the real image of the sample passes through the spatial mask to the 4f system; an optical granting, disposed on a Fourier plane in middle of the 4f system, and configured to split the real image of the sample to obtain a split real image; and an image sensor, configured to obtain the split real image to obtain an image of the sample. The present disclosure advantages of improving imaging rate in multi-spectrum fluorescence microscopy.

First claim

What is claimed is:

1. An imaging method, applied in an imaging system, wherein the system comprises: a fluorescence microscope, wherein the fluorescence microscope comprises a laser source and an objective lens, a sample is disposed on a focal plane of the objective lens, the laser source is configured to emit lasers to illuminate the sample, and the objective lens is configured to magnify the sample illuminated by the lasers to an image plane of the objective lens, to obtain a real image of the sample; a spatial mask, disposed on the image plane of the objective lens, and configured to perform mask modulation on the real image of the sample; a 4f system, disposed behind the spatial mask, wherein a beam of the real image of the sample passes through the spatial mask to the 4f system; an optical grating, disposed on a Fourier plane in middle of the 4f system, and configured to split the beam of the real image of the sample to obtain split beams of the real image; and an image sensor, configured to obtain the split beams of the real image to obtain an image of the sample, and the method comprises:
obtaining an image of the sample with the imaging system;
obtaining at least one sparse representation coefficient of the image under a preset multi-spectrum over-complete dictionary, wherein the multi-spectrum over-complete dictionary comprises a plurality of over-complete dictionaries corresponding to a plurality of spectrum bands respectively;
performing a spectrum separation on the image according to the at least one sparse representation coefficient and at least one spectrum band corresponding to the at least one sparse representation coefficient, to obtain a multi-spectrum image of the sample.

2. The method according to claim 1 , wherein obtaining at least one sparse representation coefficient of the image under a preset multi-spectrum over-complete dictionary comprises:
solving the image with K-SVD algorithm according to the preset multi-spectrum over-complete dictionary, to obtain the at least one sparse representation coefficient.

3. The method according to claim 1 , further comprising: establishing the preset multi-spectrum over-complete dictionary,
wherein establishing the preset multi-spectrum over-complete dictionary comprises:
labeling samples of different structures with different fluorescence, to obtain a plurality of multi-spectrum images;
performing learning and training on the plurality of multi-spectrum images to obtain a plurality of original over-complete dictionaries, wherein each of the plurality of original over-complete dictionaries corresponds to a spectrum band;
obtaining a point spread function of the optical grating;
performing convolution on the plurality of original over-complete dictionaries and the point spread function, to obtain the preset multi-spectrum over-complete dictionary.

4. The method according to claim 1 , wherein the sample is a fluorescence sample, the method further comprising:
magnifying the fluorescence sample by the objective lens, to obtain the real image on the focal plane of the objective lens;
reflecting a beam of the real image by a reflector to the 4f system, wherein the reflector is disposed on the focal plane of the objective lens, and the focal plane of the objective lens is a front focal plane of the 4f system;
splitting the beam of the real image by both the 4f system and the optical grating into the split beams of the real image, wherein the optical grating is disposed on a focal plane in middle of the 4f system; and
obtaining the split beams by a collecting device to obtain a multi-color fluorescence image.

5. The method according to claim 4 , further comprising:
deflecting, by the optical grating, the beam having different wavelengths with different angles respectively, to realize splitting.

6. The method according to claim 4 , wherein the collecting device realizes spectrum separation by a convolution algorithm.

Assignees

• Univ Tsinghua

Inventors

• Dai Qionghai
• Chen Xingye
• Wu Jiamin

Classifications

• G02B21/16

  adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} · CPC title

• G02B21/0032 Primary

  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 title

• H04N13/257

  Colour aspects · CPC title

• G02B21/0064

  multi-spectral or wavelength-selective arrangements, e.g. wavelength fan-out, chromatic profiling (G02B21/0076 takes precedence) · CPC title