US-12618776-B2 - Multi-fluorescence image acquisition system and method, and application

US12618776B2US 12618776 B2US12618776 B2US 12618776B2US-12618776-B2

Abstract

A system for acquiring multi-fluorescence images includes an excitation light source configured to irradiate an excitation beam onto a surface of a test sample, so that the test sample emits multi-fluorescence signals. The multi-fluorescence signals include at least two fluorescence signals. The system also includes an optical detection device configured to acquire the multi-fluorescence signals, an optical lens disposed between the optical detection device and the test sample, and a multi-band filter disposed between the optical detection device and the optical lens and/or between the test sample and the optical lens. The optical lens is configured to converge the multi-fluorescence signals emitted by the test sample to the optical detection device. The multi-band filter has at least two transmission intervals corresponding to at least two wavelength intervals of the at least two fluorescence signals. The optical detection device is configured to generate multi-fluorescence images based on the multi-fluorescence signals.

Inventors

Haohan Xia
Yanfang Chen

Assignees

SHANGHAI RUIYU BIOTECH CO., LTD.

Dates

Publication Date: 20260505
Application Date: 20231128
Priority Date: 20210528

Claims (20)

1 . A system for acquiring multi-fluorescence images, comprising an excitation light source, an optical detection device, an optical lens, and a multi-band filter; wherein the excitation light source is configured to irradiate an excitation beam onto a test sample, so that the test sample emits a first multi-fluorescence signal, the first multi-fluorescence signal including at least two fluorescence signals, and different fluorescence signals corresponding to different wavelength intervals; the optical lens is disposed between the optical detection device and the test sample, and the optical lens is configured to converge the first multi-fluorescence signal to the optical detection device; the multi-band filter is disposed between the optical detection device and the optical lens and/or between the test sample and the optical lens, the multi-band filter has at least two transmission intervals corresponding to at least two wavelength intervals, and the multi-band filter is configured to filter the first multi-fluorescence signal to generate a second multi-fluorescence signal; the optical detection device is configured to acquire the second multi-fluorescence signal and generate a multi-fluorescence image based on the second multi-fluorescence signal; the system further comprises a sample region for placing the test sample, wherein the sample region includes a base; the base is configured to place the test sample; and the base includes a first light-transmitting medium; an excitation light path is configured in a second light-transmitting medium, a refractive index of the second light-transmitting medium is not equal to a refractive index of the first light-transmitting medium, and the excitation light path is a light path through which the excitation beam is incident onto the test sample.
2 . The system of claim 1 , wherein a refractive index of the first light-transmitting medium is within a range of 1.3-1.8.
3 . The system of claim 1 , wherein the excitation light source and the optical detection device are respectively disposed on different sides of the test sample; or the excitation light source and the optical detection device are disposed on a same side of the test sample.
4 . The system of claim 1 , wherein an incident angle of the excitation beam relative to the base is configured such that the excitation beam deflects from the optical lens after being refracted by the first light-transmitting medium.
5 . The system of claim 1 , further comprising a dichroic mirror disposed between the optical lens and the multi-band filter.
6 . The system of claim 5 , wherein the dichroic mirror and the multi-band filter form a first preset angle.
7 . The system of claim 6 , wherein the first preset angle is within a range of 40°-50°.
8 . The system of claim 1 , wherein when the excitation light source is a monochromatic light source, the system further includes an excitation filter provided between the excitation light source and the test sample; and when the excitation light source is a laser light source, the system is not provided with the excitation filter.
9 . The system of claim 1 , wherein the system includes only one excitation light source.
10 . The system of claim 1 , wherein the system includes only one optical detection device.
11 . The system of claim 1 , wherein the optical lens includes an optical element or a lens group with a light converging function.
12 . The system of claim 1 , wherein the multi-band filter is a glass slide wrapped with a multi-band filter film or a glass slide coated with a multi-band filter material.
13 . A method for acquiring multi-fluorescence images, implemented based on a system for acquiring multi-fluorescence images, wherein the system comprises: an excitation light source, an optical detection device, an optical lens, and a multi-band filter; wherein the excitation light source is configured to irradiate an excitation beam onto a test sample, so that the test sample emits a first multi-fluorescence signal, the first multi-fluorescence signal including at least two fluorescence signals, and different fluorescence signals corresponding to different wavelength intervals; the optical lens is disposed between the optical detection device and the test sample, and the optical lens is configured to converge the first multi-fluorescence signal to the optical detection device; the multi-band filter is disposed between the optical detection device and the optical lens and/or between the test sample and the optical lens, the multi-band filter has at least two transmission intervals corresponding to at least two wavelength intervals, and the multi-band filter is configured to filter the first multi-fluorescence signal to generate a second multi-fluorescence signal; the optical detection device is configured to acquire the second multi-fluorescence signal and generate a multi-fluorescence image based on the second multi-fluorescence signal; the method comprises: irradiating the excitation beam to the test sample, the test sample being a measured object labeled with multiple fluorescent dyes; the system further comprises a sample region for placing the test sample, wherein the sample region includes a base; the base is configured to place the test sample; and the base includes a first light-transmitting medium; an excitation light path is configured in a second light-transmitting medium, a refractive index of the second light-transmitting medium is not equal to a refractive index of the first light-transmitting medium, and the excitation light path is a light path through which the excitation beam is incident onto the test sample.
14 . The method of claim 13 , wherein the excitation light source is a laser light source or a monochromatic light source.
15 . The method of claim 13 , wherein wavelength intervals of the excitation beam correspond to excitation peaks of the multiple fluorescent dyes.
16 . The method of claim 13 , wherein transmission intervals of the multi-band filter correspond to emission bands of the multiple fluorescent dyes of the test sample, or the transmission intervals of the multi-band filter correspond to maximum emission peaks of the multiple fluorescent dyes of the test sample.
17 . The method of claim 13 , wherein the transmission intervals of the multi-band filter do not coincide with the wavelength intervals of the excitation beam.
18 . The method of claim 13 , wherein the measured object is a cell sample, a protein sample, an antibody sample, a carbohydrate sample, or a drug sample.
19 . The method of claim 13 , wherein a position of an emission end of the excitation light source relative to the base is adjusted, so that the excitation beam emitted by the excitation light source is vertically incident onto the base.
20 . The method of claim 14 , wherein the test sample includes at least an acridine orange (AO) fluorescent dye and a propidium iodide (PI) fluorescent dye.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of International Application No. PCT/CN2022/095784, filed on May 27, 2022, which claims priority of the Chinese Patent Application No. 202110593167.3, filed on May 28, 2021, the entire content of each of which are hereby incorporated by reference. TECHNICAL FIELD The present disclosure relates to the field of in-vitro diagnostic technology, and in particular, to systems and methods for detecting multi-fluorescence signals and acquiring multi-fluorescence images and application thereof. BACKGROUND In detection or analysis applications at the biological cell level, the status of labeled cells may often be analyzed through fluorescence imaging. For example, in fluorescence imaging technology, samples containing fluorescent labels (e.g., cells labeled with fluorescent dyes, etc.) may emit fluorescence signals under the irradiation of an excitation light source, so that the device for detecting fluorescence signals may collect the fluorescence signals to generate corresponding fluorescence images, thereby analyzing the status of labeled cells based on the fluorescence images. However, in practical application, multi-fluorescence imaging, e.g., the acridine orange (AO)/propidium iodide (PI) viability detection approach, is often required to analyze different properties of fluorescent labels. The device for detecting the fluorescence signals may only acquire a single fluorescence signal in one detection. Multi-fluorescence signals may only be acquired by collecting fluorescence signals for multiple times to form multi-fluorescence images. This approach leads to a complex structure and high cost of the device for detecting the fluorescence signals, resulting in difficulty of application and promotion. Therefore, it is desirable to provide systems for detecting multi-fluorescence signals with a simple structure, low cost and capable of acquiring multi-fluorescence images simultaneously. SUMMARY One of the embodiments of the present disclosure provides a system for acquiring multi-fluorescence images. The system may comprise an excitation light source, an optical detection device, an optical lens, and a multi-band filter. The excitation light source may be configured to irradiate an excitation beam onto a test sample, so that the test sample may emit a first multi-fluorescence signal. The first multi-fluorescence signal may include at least two fluorescence signals, and different fluorescence signals may correspond to different wavelength intervals. The optical lens may be disposed between the optical detection device and the test sample. The optical lens may be configured to converge the first multi-fluorescence signal to the optical detection device. The multi-band filter may be disposed between the optical detection device and the optical lens and/or between the test sample and the optical lens. The multi-band filter may have at least two transmission intervals corresponding to at least two wavelength intervals. The multi-band filter may be configured to filter the first multi-fluorescence signal to generate a second multi-fluorescence signal. The optical detection device may be configured to acquire the second multi-fluorescence signal and generate a multi-fluorescence image based on the second multi-fluorescence signal. In some embodiments, the system for acquiring the multi-fluorescence images may further include a sample region for placing the test sample. In some embodiments, a refractive index of a first light-transmitting medium may be within a range of 1.3-1.8. In some embodiments, the excitation light source and the optical detection device may be respectively disposed on different sides of the test sample; or the excitation light source and the optical detection device may be disposed on a same side of the test sample. In some embodiments, an incident angle of the excitation beam relative to a base may be configured such that the excitation beam deflects from the optical lens after being refracted by the first light-transmitting medium. In some embodiments, an excitation light path may be configured in a second light-transmitting medium, and a refractive index of the second light-transmitting medium may not be equal to a refractive index of the first light-transmitting medium. The excitation light path may be a light path through which the excitation beam is incident onto the test sample. In some embodiments, the system for acquiring the multi-fluorescence images may further include a dichroic mirror disposed between the optical lens and the multi-band filter. In some embodiments, the dichroic mirror and the multi-band filter may form a first preset angle. In some embodiments, the first preset angle may be within a range of 40°-50°. In some embodiments, when the excitation light source is a monochromatic light source, the system for acquiring the multi-fluorescence images may further include an excitation filter provided between t