CN115710547A

CN115710547A - PCR optical path system based on diffraction optical element structure

Info

Publication number: CN115710547A
Application number: CN202110973836.XA
Authority: CN
Inventors: 朱守明; 张鑫
Original assignee: Anhui Wanyi Science and Technology Co Ltd
Current assignee: Anhui Wanyi Science and Technology Co Ltd
Priority date: 2021-08-23
Filing date: 2021-08-23
Publication date: 2023-02-24

Abstract

The invention discloses a PCR light path system based on a diffractive optical element structure, which comprises a laser module, a Dammann grating, a light splitting module, a focusing lens, a 96-hole plate and a fluorescence camera. Laser emitted by the laser module is divided into 96 light spots with consistent intensity through the Dammann grating, the 96 light spots are reflected to the mirror surface of the focusing mirror through the light splitting module, then the exciting light emitted by the light splitting module is collimated and emitted into the 96 pore plate through the focusing mirror, so that a fluorescent sample in the 96 pore plate is excited to emit fluorescence, and the fluorescence is finally collected by the fluorescent camera. The invention uses the laser module as a light source, does not need to excite an optical filter, directly generates 96 beams of laser with uniform intensity by using the Dammann grating, has high excitation light intensity and no edge effect because the light spot position corresponds to the position of a 96-hole plate, does not need scanning, and has quick and rapid detection.

Description

PCR optical path system based on diffraction optical element structure

Technical Field

The invention relates to a PCR system, in particular to a PCR optical path system based on a diffractive optical element structure.

Background

At present, most of the conventional PCR light paths use a tungsten halogen lamp light source or an LED light source, which has a broad spectrum of wavelength and needs an excitation filter to obtain the required narrow-band excitation light. The PCR light path is mostly in a fiber structure or a mode of projecting a large light spot. The optical fiber method is generally to make the optical fiber head face to the reagent tube containing the sample, fix the optical fiber head on the mechanical part, and control the mechanical part to move on the slide rail through the motor to realize the scanning of a plurality of reagent tubes, or control the mechanical part carrying the sample reagent tube to move through the motor to scan, however, as is well known, the PCR system consumes a long time during scanning; the mode of projecting the large light spot is to put the whole reagent tube filled with the sample under the large light spot for fluorescence excitation without scanning, but the mode has higher requirement on the power of a light source and can be realized by a high-power light source, and then the CCD camera is utilized to collect fluorescence.

Disclosure of Invention

In order to solve the above-mentioned deficiencies in the prior art, the present invention provides a PCR optical path system based on a diffractive optical element structure, which can eliminate the scanning method for detecting a fluorescent sample, and the detection is rapid and quick.

The technical scheme adopted by the invention for solving the technical problems is as follows: a PCR optical path system based on a diffractive optical element structure, comprising:

the laser module is used for emitting laser;

the Dammann grating is arranged on a rear-stage light path of the laser module, so that laser emitted by the laser module is divided into 96 beams of light through the Dammann grating;

the light splitting module is arranged on a rear-stage light path of the Dammann grating, so that laser split by the Dammann grating is emitted to a fluorescent sample to be detected through the light splitting module;

the focusing mirror is arranged on a rear-stage light path of the light splitting module;

the 96-hole plate is arranged in the rear-stage optical path of the focusing mirror, and the 96-hole plate is positioned at the focal plane position of the focusing mirror;

and the fluorescence camera is arranged on a post-stage optical path of the 96-well plate, so that a fluorescence signal excited in the 96-well plate is received by the fluorescence camera.

Optionally, the number of the laser modules is at least two, and the laser wavelengths emitted by at least two of the laser modules are respectively adapted to the corresponding fluorescence samples.

Optionally, optical axes of the laser beams emitted by at least two laser modules are coupled to the same optical axis through first dichroic mirrors respectively;

the number of the first dichroic mirrors is one less than that of the laser modules.

Optionally, the light splitting module includes a plurality of second dichroic mirrors and a plurality of emission filters;

the number of the second dichroic mirrors and the number of the emission optical filters are respectively consistent with the number of the laser modules, so that the laser modules correspond to the corresponding second dichroic mirrors and the emission optical filters in the light splitting modules.

Optionally, the hole center-to-center spacing of the 96-well plate is 9mm.

Optionally, the dammann grating is made of one or more of fused silica, polycarbonate, polymethyl methacrylate, and zinc selenide.

By adopting the technical scheme, compared with the prior art, the invention has the following beneficial effects:

1. the invention adopts the laser module as the light source, the light intensity of the laser module is very high, the wavelength width is very narrow, only a few nanometers, and an excitation optical filter is not needed, so that the cost of a PCR instrument is saved;

2. the invention uses the Dammann grating to split the incident exciting light into 96 beams, 96 light spots are obtained, the intensity of the 96 light spots is consistent, the influence of the inconsistent intensity of the exciting light on the fluorescence intensity is avoided, the positions of the light spots correspond to the hole positions of a 96 hole plate, no angle deflection exists, and no edge effect exists;

3. the invention abandons the mode of detecting the fluorescent sample by adopting a scanning mode in the traditional PCR instrument, so that the detection is rapid and quick.

Drawings

FIG. 1 is a schematic diagram of an optical path system of the present invention;

FIG. 2 is a schematic cross-sectional view of a Dammann grating of the present invention;

FIG. 3 is a schematic diagram showing the distribution of 96 wells in the 96-well plate of the present invention.

Detailed Description

The present application will be described in further detail with reference to the following drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the relevant invention and not restrictive of the invention. It should be noted that, for convenience of description, only the portions related to the present invention are shown in the drawings.

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present application will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.

As shown in fig. 1, the present invention discloses a PCR optical path system based on a diffractive optical element structure, which comprises: the device comprises a laser module, a Dammann grating 4, a light splitting module, a focusing mirror 7, a 96-hole plate 8 and a fluorescence camera 9.

And the laser module is used for emitting laser so that the fluorescence sample can be excited to emit fluorescence through the laser with corresponding wavelength. In the present invention, a plurality of laser modules may be provided to enable the plurality of laser modules to emit laser beams with different wavelengths, so as to meet the requirement of multi-channel selection, for example, in an embodiment of the present invention, three laser modules may be provided, which are the first laser module 1, the second laser module 2, and the third laser module 3. When arranging first laser module 1, second laser module 2 and third laser module 3, in order to practice thrift the space in the PCR instrument, make the outgoing optical axis of second laser module 2, third laser module 3 perpendicular with the outgoing optical axis of first laser module 1, wherein, the outgoing optical axis of first laser module 1 is the primary optical axis. Then, set up first dichroic mirror 5 in the emergent light path of second laser module 2, third laser module 3 respectively, the face of two first dichroic mirrors 5 matches with the wavelength of second laser module 2, third laser module 3 respectively to make the laser that second laser module 2 sent be coupled to the primary optical axis under the effect of corresponding first dichroic mirror 5, make it shoot out along the primary optical axis, third laser module 3 is the same, no longer describe repeatedly. In addition, the laser wavelength of first laser module 1, second laser module 2 and third laser module 3 grows gradually, and the laser module of long wavelength distributes in the light path of back level promptly. The laser emitted by the first laser module 1, the second laser module 2 and the third laser module 3 is a point laser which can be emitted in a collimating way, and the bandwidth is narrow and only a few nanometers, so that an excitation filter is not required to be added.

The Dammann grating 4 is arranged on a rear-stage light path of the laser module, so that laser emitted by the laser module is diffracted by the Dammann grating 4 and then is divided into 96 beams of light, the laser forms 96 light spots, and the intensity of the 96 light spots is uniform. The cross section of the dammann grating 4 is shown in fig. 2, and the dammann grating 4 is made of one or more of fused silica, polycarbonate, polymethyl methacrylate and zinc selenide.

And the light splitting module is arranged on a rear-stage light path of the Dammann grating 4, so that the laser split by the Dammann grating 4 is emitted to the fluorescent sample to be detected through the light splitting module. Specifically, the spectral module includes second dichroic mirror 5 and emission filter 6, wherein, every second dichroic mirror 5 corresponds an emission filter 6, that is to say, a second dichroic mirror 5 and an emission filter 6 constitute a spectral module, and the quantity of spectral module is unanimous with the quantity of laser module, make every laser module corresponding with corresponding spectral module, second dichroic mirror 5 and emission filter 6 set up according to the laser module of corresponding wavelength promptly, when switching the testing channel, also need switch corresponding spectral module, consequently, a plurality of spectral module are integrated in a rotary platform, switch the spectral module through rotatory mode.

And the focusing mirror 7 is arranged on a rear-stage light path of the light splitting module and is used for collimating the laser light reflected by the second dichroic mirror 5.

And the 96-hole plate 8 is arranged in the rear-stage optical path of the focusing mirror 7, and the 96-hole plate 8 is positioned at the focal plane position of the focusing mirror 7. The 96 wells of the 96-well plate 8 are each loaded with a fluorescent sample. In the 96-well plate 8, the distance between the centers of two adjacent holes is 9mm, the holes are distributed as shown in fig. 3, the hole sites of the 96-well plate 8 correspond to 96 laser spots respectively, and the 96 laser spots are vertically incident into 96 holes of the 96-well plate 8 respectively.

And a fluorescence camera 9 provided in a subsequent optical path of the 96-well plate 8 so that a fluorescence signal excited in the 96-well plate 8 is collected by the fluorescence camera. In the process of collecting fluorescence, stray light of the fluorescence is filtered by the emission filter 6, so that the fluorescence with corresponding wavelength penetrates through the emission filter 6 and is collected by the fluorescence camera 9.

The foregoing description is only exemplary of the preferred embodiments of the application and is illustrative of the principles of the technology employed. It will be appreciated by a person skilled in the art that the scope of the invention as referred to in the present application is not limited to the embodiments with a specific combination of the above-mentioned features, but also covers other embodiments with any combination of the above-mentioned features or their equivalents without departing from the inventive concept. For example, the above features may be replaced with (but not limited to) features having similar functions disclosed in the present application.

Besides the technical features described in the specification, other technical features are known to those skilled in the art, and are not described in detail herein in order to highlight the innovative features of the present invention.

Claims

1. A PCR optical path system based on a diffraction optical element structure is characterized by comprising:

the laser module is used for emitting laser;

2. The PCR optical path system based on the diffractive optical element structure of claim 1, wherein the number of the laser modules is at least two, and the laser wavelengths emitted by at least two of the laser modules are respectively adapted to the corresponding fluorescence samples.

3. The PCR optical path system based on the diffraction optical element structure as claimed in claim 2, wherein the optical axes of the laser beams emitted by at least two laser modules are respectively coupled into the same optical axis through a first dichroic mirror;

4. The diffractive optical element structure-based PCR optical path system according to claim 3, wherein the light splitting module comprises a plurality of second dichroic mirrors and a plurality of emission filters;

5. The diffraction optical element structure-based PCR optical path system of claim 4, wherein the hole center-to-center spacing of the 96-well plate is 9mm.

6. The PCR optical path system based on the diffractive optical element structure of claim 5, wherein the Dammann grating is made of one or more of fused silica, polycarbonate, polymethyl methacrylate and zinc selenide.