IMAGING SYSTEM FOR MULTIPLEX DETECTION OF BIOMOLECULES

Abstract

An imaging system for multiplex detection of biomolecules, including at least a first optical path and a second optical path; the first optical path is from an excitation light source to an objective lens, and a filter set is provided between the excitation light source and the objective lens and is configured to guide excitation light from the excitation light source to the objective lens; the second optical path is from the objective lens to an image acquisition device, a reflector is provided between the objective lens and the image acquisition device and is configured to project the fluorescence received by the objective lens to the image acquisition device; the excitation light source includes at least a first excitation light source used for excitation of fluorescent signals and a second excitation light source used for excitation of light-initiated chemiluminescence signals The system simultaneously realizes fluorescence and light-initiated chemiluminescence imaging.

Description

Claims

1. An imaging system for multiplex detection of biomolecules, wherein the imaging system simultaneously realizes fluorescence imaging and light-initiated chemiluminescence imaging.

2. An imaging system for multiplex detection of biomolecules, wherein the imaging system comprises at least a first optical path and a second optical path;the first optical path is from an excitation light source (1) to an objective lens (15), a filter set (13) is provided between the excitation light source and the objective lens, and the filter set (13) is configured to guide excitation light from the excitation light source to the objective lens;the second optical path is from the objective lens (15) to an image acquisition device (2), a reflector (16) is provided between the objective lens (15) and the image acquisition device (2), and the reflector (16) is configured to project fluorescence received by the objective lens (15) to the image acquisition device (2);the excitation light source (1) comprises at least a first excitation light source (11) and a second excitation light source (12), wherein the first excitation light source (11) is used for excitation of fluorescent signals, and the second excitation light source (12) is used for excitation of light-initiated chemiluminescence signals.

3. The imaging system according to claim 2, wherein the imaging system further comprises a workstation (3) and a light source switch (17), the workstation is connected in communication with the image acquisition device (2), and the light source switch (17) is connected in communication with the excitation light source (1) and the workstation (3).

4. The imaging system according to claim 3, wherein the workstation comprises a control center (31) and a trigger control box (32); wherein the trigger control box (32) is connected in communication with the excitation light source (1), the light source switch (17) and the image acquisition device (2); wherein the control center (31) is connected in communication with the image acquisition device (2) and the trigger control box (32).

5. The imaging system according to claim 2, wherein the imaging system comprises an ordinary fluorescence imaging mode and a time-resolved fluorescence mode; in the ordinary fluorescence imaging mode, light emission of the excitation light source is synchronized with exposure of the image acquisition device; in the time-resolved fluorescence mode, light emission of the excitation light source is not synchronized with exposure of the image acquisition device.

6. The imaging system according to claim 2, wherein the imaging system further comprises one or more of the following features: a. the first excitation light source (11) or the second excitation light source (12) is selected from one or more of a laser, a xenon lamp, a mercury lamp, a halogen lamp, and a light emitting diode;b. the image acquisition device (2) is selected from one or more of charge-coupled device (CCD), electron-multiplying charge-coupled device (EMCCD), complementary metal-oxide-semiconductor image sensor (CMOS), and scientific complementary metal-oxide-semiconductor image sensor (sCMOS);c. the filter set (13) comprises at least a first filter subset, a second filter subset and a third filter subset, and each filter subset comprises an excitation filter, an emission filter and a dichroic mirror;d. the filter set (13) is detachable.

7. The imaging system according to claim 6, wherein the imaging system further comprises one or more of the following features: (1) the first filter subset comprises: excitation filter 488/15 nm, dichroic mirror 495 nm longpass, emission filter 535/23 nm;(2) the second filter subset comprises: excitation filter 488/15 nm, dichroic mirror 495 nm longpass, emission filter 600/40 nm;(3) the third filter subset comprises: excitation filter 680/13 nm, dichroic mirror 653 nm shortpass, emission filter 615/20 nm.

8. An use of the imaging system according to claim 2 in the filed of multiplex detection of biomolecules.

9. An imaging method for multiplex detection of biomolecules, wherein the imaging method uses the the imaging system according to claim 2, wherein the imaging method at least comprises: 1) turning on the first excitation light source of the imaging system, so that light emitted by the first excitation light source passes through the filter set and then the objective lens to reach a sample for imaging, thereby obtaining a fluorescence image of the sample;2) switching to the second excitation light source, so that light emitted by the second excitation light source passes through the filter set and then the objective lens to reach the sample for imaging, thereby obtaining a light-initiated chemiluminescence image of the sample.

10. The imaging method according to claim 9, wherein the imaging method further comprises: changing parameters of the filter set and repeating the operations in 1) to obtain multiple fluorescence images of the sample.

11. The imaging method according to claim 9, wherein after switching to the second excitation light source in operation 2), a time-resolved fluorescence mode is turned on to obtain the light-initiated chemiluminescence image of the sample.

12. An image analysis method for multiplex detection of biomolecules, comprising: S1: matching a fluorescence image of a detection solution with a light-initiated chemiluminescence image of the detection solution according to light spot positions, to obtain a fluorescence signal and a light-initiated chemiluminescence signal corresponding to each light spot, wherein the fluorescence image and the light-initiated chemiluminescence image are obtained by the imaging method according to claim 9;S2: performing cluster classification on the fluorescence signals, and obtaining an average light-initiated chemiluminescence intensity corresponding to each fluorescence classification;S3: determining the content of each molecule to be tested in the detection solution according to the different classifications of the fluorescent signals and the average light-initiated chemiluminescence intensity corresponding to each fluorescence classification.

13. A computer-readable storage medium having a computer program stored thereon, wherein the program implements the operations of the image analysis method according to claim 12 when executed by a processor.