CN217265753U

CN217265753U - Real-time fluorescent quantitative PCR detection system for multi-flux samples

Info

Publication number: CN217265753U
Application number: CN202221038019.1U
Authority: CN
Inventors: 吉俊利; 周志图
Original assignee: Bioteke Corp
Current assignee: Bioteke Corp
Priority date: 2022-04-29
Filing date: 2022-04-29
Publication date: 2022-08-23
Anticipated expiration: 2032-04-29

Abstract

The utility model discloses a real-time fluorescence quantitative PCR detecting system of many flux samples, include: the excitation light path comprises an excitation light source, an optical fiber, a light guide rod and an anti-distortion lens group; the light emitted by the excitation light source enters the light guide rod after passing through the optical fiber and finally enters the distortion elimination lens group; and a detection optical path comprising a sample carrier on an optical path of the excitation optical path, the sample carrier being located downstream of the anamorphic lens group, and an imaging device disposed toward the sample carrier to capture an image of the sample on the sample carrier. The utility model discloses technical scheme can improve the precision that multi-flux sample PCR detected.

Description

Real-time fluorescent quantitative PCR detection system for multi-flux samples

Technical Field

The utility model relates to a PCR detects technical field, in particular to real-time fluorescence quantitative PCR detecting system of many flux samples.

Background

The global prevalence of new crown epidemics makes nucleic acid detection a detection project that everyone participates in. After collecting a body fluid sample of a subject, a tester amplifies a specific DNA (deoxyribose nucleic Acid) sequence by PCR (Polymerase Chain Reaction). If the DNA sequence is present in the sample, a specific fluorescent probe can be injected into the sample, and the fluorescent probe can only bind to the DNA sequence. After the fluorescent probe is combined with a specific DNA sequence, the fluorescent marker can fall off from the probe. Since the detached fluorescent label emits fluorescence when excited by a specific light, detection of fluorescence indicates that the sample contains the DNA sequence.

By adopting the PCR detection of the multi-flux samples, multi-component samples can be detected at one time, and the detection efficiency is improved. But the efficiency is improved and the detection accuracy is reduced.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a real-time fluorescence quantitative PCR detecting system of many flux samples aims at improving the precision that the PCR of many flux samples detected.

In order to achieve the above object, the present invention provides a real-time fluorescence quantitative PCR detection system of multi-flux sample, comprising:

the excitation light path comprises an excitation light source, an optical fiber, a light guide rod and an anti-distortion lens group; the light emitted by the excitation light source enters the light guide rod after passing through the optical fiber and finally enters the distortion elimination lens group; and

a detection optical path comprising a sample carrier on an optical path of the excitation optical path, the sample carrier being located downstream of the anamorphic lens group, and an imaging device disposed toward the sample carrier to capture a sample image on the sample carrier.

Optionally, the excitation light sources are laser light sources and are arranged in multiple groups, each group has one or more laser light sources, the laser light sources in the same group have the same wavelength, and the laser light sources in different groups have different wavelengths.

Optionally, the optical fiber is a Y-shaped optical fiber, and each laser light source is coupled into an incident end of one Y-shaped optical fiber; the power supplies of different groups of the laser light sources are mutually independent and can be independently switched on and off.

Optionally, the sample carrier further includes a plurality of dichroic mirrors provided in one-to-one correspondence with the plurality of groups of laser light sources, and a dichroic mirror selected to correspond to an active group of laser light sources is provided between the anamorphic lens group and the sample carrier, the dichroic mirror being configured to reflect laser light emitted from the anamorphic lens group toward the sample carrier.

Optionally, the imaging device is located on a side of the dichroic mirror remote from the sample carrier, the imaging device being able to capture an image of the sample carrier through the dichroic mirror.

Optionally, the exit of the light guide rod has the same shape as the edge of the sample carrier.

Optionally, the distortion elimination lens group is composed of two plano-convex lenses and a biconvex lens, which are coaxially disposed, the biconvex lens is located between the two plano-convex lenses, and convex surfaces of the two plano-convex lenses are respectively opposite to two sides of the biconvex lens.

Optionally, the detection optical path further includes a plurality of optical filters disposed at an incident end of the imaging device, one optical filter corresponds to one group of the laser light sources, the optical filter can filter laser light emitted by the corresponding laser light source, and the optical filter can transmit fluorescence emitted by the sample excited by the corresponding laser light source.

Optionally, the excitation light path further includes a rod mirror disposed between the laser light source and the optical fiber.

Optionally, the rod mirror includes a first rod mirror and a second rod mirror, and the first rod mirror and the second rod mirror are both coaxially disposed with the excitation light source; the length direction of the first rod mirror is vertical to the fast axis of the laser; the length direction of the second rod mirror is perpendicular to the slow axis of the laser.

The utility model discloses technical scheme is through adopting the distortion elimination battery of lens for the light beam of exciting light intensity is more even in the cross direction, makes the illumination intensity that each sample received more close, so contrast every sample the fluorescence intensity's that sends difference after, more can reflect the probe and receive the combination condition of the DNA sequence that detects, improves the detection precision from this.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the structures shown in the drawings without creative efforts.

FIG. 1 is a schematic structural diagram of an embodiment of the real-time fluorescence quantitative PCR detection system for multi-flux samples according to the present invention;

FIG. 2 is an enlarged view of a portion of FIG. 1 at A;

FIG. 3 is a schematic diagram of a Y-shaped optical fiber according to the embodiment of FIG. 1;

FIG. 4 is a simulated view of the rod mirror to laser coupling of the embodiment of FIG. 1;

FIG. 5 is a simulated view of the effect of the light guide rod and the anamorphic lens set on the laser path of the embodiment of FIG. 1;

FIG. 6 is a graph showing the light intensity distribution of the embodiment of FIG. 1 at the light spot and at two positions of the light spot on the sample carrier.

The reference numbers illustrate:

reference numerals	Name (R)	Reference numerals	Name (R)
				10	Light guide rod	60	Y-shaped optical fiber
20	Distortion eliminating lens group	61	Y-shaped optical fiber incident end
				21	Incident plano-convex lens	62	Y-shaped optical fiber emergent end
22	Emergent plano-convex lens	70	Dichroic mirror
				23	Biconvex lens	80	Optical filter
30	Sample carrier	91	First rod mirror
				40	Image forming apparatus	92	Second rod mirror
50	Laser light source

The objects, features and advantages of the present invention will be further described with reference to the accompanying drawings.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

It should be noted that all the directional indicators (such as upper, lower, left, right, front and rear … …) in the embodiment of the present invention are only used to explain the relative position relationship between the components, the motion situation, etc. in a specific posture (as shown in the drawings), and if the specific posture is changed, the directional indicator is changed accordingly.

In addition, the descriptions related to "first", "second", etc. in the present invention are for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicit ly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the feature. In addition, the technical solutions in the embodiments may be combined with each other, but it must be based on the realization of those skilled in the art, and when the technical solutions are contradictory or cannot be realized, the combination of the technical solutions should not be considered to exist, and is not within the protection scope of the present invention.

The utility model provides a real-time fluorescence quantitative PCR detecting system of many flux samples.

Referring to fig. 1 to 3, in an embodiment of the present invention, the multi-throughput sample real-time fluorescence quantitative PCR detection system includes:

the excitation light path comprises an excitation light source, an optical fiber, a light guide rod 10 and an anti-distortion lens group 20; the light emitted by the excitation light source enters the light guide rod 10 after passing through the optical fiber and finally enters the distortion elimination lens group 20; and

a detection optical path comprising the sample carrier 30 and an imaging device 40, the sample carrier 30 being in the optical path of the excitation optical path, the sample carrier 30 being downstream of the anamorphic lens group 20, the imaging device 40 being disposed towards the sample carrier 30 to capture an image of the sample on the sample carrier 30.

The excitation light path is used for providing excitation light for the sample, and the detection light path is used for recording the intensity and the time change of fluorescence emitted by the sample. Wherein the sample carrier 30 can carry a plurality of samples, such that when the excitation light generated by the excitation light path is irradiated onto the sample carrier 30, the plurality of samples can be excited to emit fluorescence simultaneously; meanwhile, the imaging device 40 can capture the luminescence of all samples at the same time, so that the purpose of multi-flux sample detection can be achieved. Three factors can influence the luminous intensity of the sample, namely the measurement of the fluorescent probe injected into the sample, the concentration of the target DNA sequence in the sample and the intensity of the exciting light. In order to quantitatively express the concentration of the target DNA in the sample by the intensity of fluorescence emitted from the sample, a controlled variable method is used, that is, the amount of fluorescence probe injected into the sample is the same in each sample; the intensity of the excitation light impinging on each sample was the same. With this arrangement, the intensity of the fluorescence emitted from the sample is a function of the concentration of the target DNA sequence in the sample, and quantitative measurement of the target DNA concentration can be achieved.

In order to make the intensity of shining the exciting light on every sample the same, the utility model discloses an in this embodiment, be provided with a light guide rod 10, after the line beam followed optic fibre outgoing, through this light guide rod 10, can turn into the light beam that cross sectional shape is the same with light guide rod 10 exit end cross sectional shape, and because the light beam is multiple reflection in light guide rod 10, known from the fresnel equation, even the light beam of incidenting is coherent light, the light beam of outgoing also can turn into incoherent light. This may make the properties of the excitation light more uniform.

In order to make the intensity of the exciting light irradiated on each sample the same, the embodiment of the utility model is also provided with a distortion elimination lens group 20, the distortion elimination lens group 20 can locally shape the light beam emitted from the light guide rod 10, and the divergence angle at the position with larger light intensity on the cross section of the light beam is increased; the divergence angle at which the light intensity is weaker in the cross section of the light beam is reduced, so that the light beam exiting the anamorphic lens group 20 is more uniform in cross section. Through the arrangement, the intensity of the exciting light irradiated on each sample is more consistent, and the detection precision is improved.

Referring to fig. 1, the excitation light sources are optionally laser light sources 50, and are arranged in multiple groups, each group having one or more laser light sources 50, the laser light sources 50 in the same group have the same wavelength, and the laser light sources 50 in different groups have different wavelengths. Only light of a particular wavelength can excite a particular fluorescent label to fluoresce. The excitation light source uses a light source with a wide wavelength, and in order to prevent light with a wavelength other than a specific wavelength from affecting the observation effect, an optical filter 80 may be added to filter out light with a desired wavelength. Since light outside a specific wavelength also needs to consume energy to be generated, the required power of the excitation light source needs to be sufficiently large. Since laser monochromaticity is good, the laser light source 50 can also be selected as an excitation light source, and the light filter 80 is not needed at this time, and a light source with lower power can be used. The excitation light source frequencies required by different fluorescent markers are different, and if the excitation light source with a wide wavelength range is used, different optical filters 80 are used for different fluorescent markers; if the laser is used as the excitation light source, the light sources are arranged in groups aiming at different fluorescent markers, and the laser wavelength of each group is different and is respectively aiming at different fluorescent markers.

Referring to fig. 3, optionally, the optical fibers are Y-shaped optical fibers 60, and each laser light source 50 is coupled into the incident end of one Y-shaped optical fiber 60; the power supplies of different groups of laser light sources 50 are independent of each other and can be opened and closed independently. After the arrangement, the Y-shaped optical fiber 60 has only one Y-shaped optical fiber emitting end 62, and in order to obtain the emitting light with a single wavelength, only the power supplies of the laser light sources 50 need to be managed in groups, and when the laser light with a certain wavelength is needed, the power supplies of the corresponding group of laser light sources 50 are turned on, and the power supplies of the other laser light sources 50 are turned off.

Referring to fig. 1, further, the present embodiment further includes a plurality of dichroic mirrors 70 disposed in one-to-one correspondence with the plurality of groups of laser light sources 50, the dichroic mirror 70 corresponding to the group of laser light sources 50 in use being selected to be disposed between the anamorphic lens group 20 and the sample carrier 30, the dichroic mirror 70 being configured to reflect the laser light emitted from the anamorphic lens group 20 toward the sample carrier 30. The dichroic mirror 70 has optical characteristics of having a high reflectance for light of a specific frequency band and a high transmittance for light energy of the remaining frequency bands. The dichroic mirrors 70 are arranged in one-to-one correspondence with the plurality of groups of laser light sources 50, and each dichroic mirror 70 is arranged to have a high reflectivity for light emitted by the corresponding laser light source 50; the transmittance of the fluorescence emitted by the fluorescent probe excited by the corresponding laser light source 50 is high. When the pattern of the sample carrier 30 is imaged through the dichroic mirror 70 in this way, the interference of the laser light is reduced. Without the dichroic mirror 70, the laser light would need to be incident perpendicularly on the sample carrier 30 in order to achieve a uniform intensity of light that would impinge on each sample because the laser light would have a certain divergence angle before impinging on the sample carrier 30, and without the dichroic mirror 70, the excitation light path would encroach on the space perpendicular to the sample carrier 30, and the sample carrier 30 would not be imaged in a direction perpendicular to the sample carrier 30. The dichroic mirror 70 changes the orientation of the excitation light path such that the sample carrier 30 can be photographed in a direction perpendicular to the sample carrier 30.

Referring to fig. 1, optionally, the imaging device 40 is located on a side of the dichroic mirror 70 remote from the sample carrier 30, the imaging device 40 being able to capture an image of the sample carrier 30 through the dichroic mirror 70. The imaging device 40, as described above, may be positioned perpendicular to the sample carrier 30 and may be used to photograph the sample carrier 30, which may be advantageous to restore the full appearance of the sample in the sample carrier 30. In addition, the dichroic mirror 70 filters out the reflected light of the laser light on the sample carrier 30, which is more advantageous for the imaging device 40 to shoot.

Referring to fig. 1, optionally, the shape of the exit of the light guide wand 10 is the same as the shape of the edge of the sample carrier 30. The light guide rod 10 can expand the incident light beam into emergent light having the same cross-sectional shape as the exit. If the exit of the light guide rod 10 has a different shape from the edge of the sample holder 30, the divergent angle of the outgoing light from the light guide rod 10 needs to be increased and/or the distance between the sample holder 30 and the dichroic mirror 70 needs to be increased by the anamorphic lens group 20 so that the excitation light can cover all the samples on the sample holder 30. If the exit of the light guide wand 10 has the same shape as the edge of the sample carrier 30, the excitation light can be minimally radiated away from the sample site while covering all of the sample.

Referring to fig. 1, 5 and 6, the anamorphic lens group 20 is alternatively composed of two plano-convex lenses and a biconvex lens 23 coaxially disposed, with the biconvex lens 23 being located between the two plano-convex lenses, the convex surfaces of the two plano-convex lenses being respectively opposite to both sides of the biconvex lens 23. The plano-convex lens on the incident light side of the distortion elimination lens group 20 is an incident plano-convex lens 21; the plano-convex lens on the outgoing light side is an outgoing plano-convex lens 22. The incident plano-convex lens 21 functions to focus the outgoing light from the light guide rod 10, and the lenticular lens 23 and the outgoing plano-convex lens 22 function to remove distortion generated by the incident plano-convex lens 21. The effect of the above arrangement on the light path is simulated as shown in figure 5. At this time, simulation of the excitation light intensity distribution at the sample carrier 30, which shows the laser intensity uniformity of 93% or more, is shown in fig. 6. It should be noted that, in fig. 6, the part in the left gray-scale diagram is the light spot shape of the present invention, the upper curve on the right side is the light intensity distribution curve of the light spot on the central line (i), and the lower curve on the right side is the light intensity distribution curve of the light spot on the diagonal (ii).

Referring to fig. 1, further, the embodiment of the present invention further includes a plurality of optical filters 80 disposed at the incident end of the imaging device 40, where one optical filter 80 corresponds to one group of the laser light sources 50, the optical filter 80 can filter the laser light emitted from the corresponding laser light source 50, and the optical filter 80 can transmit the fluorescence emitted from the corresponding laser light source 50. This further filters out the reflected light of the laser light source 50 at the sample carrier 30 that escapes the dichroic mirror 70. In addition, since the filter 80 has a high transmittance only for the fluorescence emitted from the sample, it can also filter the stray light from the environment.

Referring to fig. 2, further, this embodiment of the present invention further includes a rod mirror disposed between the laser light source 50 and the optical fiber. The rod mirror can focus light with the light propagation direction on the optical axis in the direction perpendicular to the length direction of the rod mirror; but the rod mirror does not change the characteristic that light whose propagation direction is on its optical axis is parallel to the length direction of the rod mirror. The laser light emitted from the laser light source 50 is a gaussian beam, and the cross section of the gaussian beam is elliptical. If a gaussian beam is coupled into an optical fiber, the convergence angles required by the fast axis direction and the slow axis direction of the gaussian beam are different, and the rod mirror can just provide the convergence angles in two different directions.

Referring to fig. 2 and 4, optionally, the rod mirrors include a first rod mirror 91 and a second rod mirror 92, and the first rod mirror 91 and the second rod mirror 92 are both disposed coaxially with the excitation light source; the length direction of the first rod mirror 91 is perpendicular to the fast axis of the laser; the second rod mirror 92 has a length direction perpendicular to the slow axis of the laser. By so doing, the laser light can be focused at a suitable point that is sufficiently close to the Y-fiber entrance end 61 that the laser light can enter the Y-fiber 60; meanwhile, the divergence angle of the point light source taking the point as the starting point can be matched with the incident angle of the Y-shaped optical fiber 60, so that the laser can generate total reflection in the optical fiber after being incident into the Y-shaped optical fiber 60. In addition, the rod lens is a conventional optical device, and rod lenses with various focal lengths are easily obtained and have low cost. Therefore, if the rod mirror of the embodiment is damaged in the using process, the rod mirror is easy to replace. If the laser source 50 is replaced with a new laser source, which results in a change in the angle of divergence of the laser, it is also easier to redesign the rod lens to couple the laser into the fiber.

The above only is the preferred embodiment of the present invention, not so limiting the patent scope of the present invention, all under the inventive concept of the present invention, the equivalent structure transformation made by the contents of the specification and the drawings is utilized, or the direct/indirect application in other related technical fields is included in the patent protection scope of the present invention.

Claims

1. A real-time fluorescent quantitative PCR detection system of a multi-flux sample is characterized by comprising:

a detection optical path comprising a sample carrier in the optical path of the excitation optical path, the sample carrier being downstream of the anamorphic lens group, and an imaging device disposed toward the sample carrier to capture an image of a sample on the sample carrier.

2. The real-time fluorescent quantitative PCR detection system of claim 1, wherein the excitation light source is a laser light source and is divided into a plurality of groups, each group has one or more laser light sources, the laser light sources in the same group have the same wavelength, and the laser light sources in different groups have different wavelengths.

3. The real-time fluorescent quantitative PCR detection system of multi-throughput samples of claim 2, wherein the optical fibers are Y-shaped optical fibers, and each of the laser light sources is coupled into an incident end of one of the Y-shaped optical fibers; the power supplies of different groups of the laser light sources are mutually independent and can be independently switched on and off.

4. The real-time quantitative PCR detection system for fluorescence of multi-throughput samples of claim 3, further comprising a plurality of dichroic mirrors disposed in one-to-one correspondence with a plurality of sets of said laser light sources, wherein a dichroic mirror selected to correspond to an active set of said laser light sources is disposed between said anamorphic lens group and said sample carrier, said dichroic mirror configured to reflect laser light exiting said anamorphic lens group toward said sample carrier.

5. The system for real-time quantitative PCR detection of multiple-throughput samples according to claim 4, wherein the imaging device is located on a side of the dichroic mirror remote from the sample carrier, the imaging device being capable of capturing an image of the sample carrier through the dichroic mirror.

6. The multi-throughput sample real-time fluorescent quantitative PCR detection system of claim 1, wherein the shape of the exit of the light guide wand is the same as the edge shape of the sample carrier.

7. The real-time fluorescent quantitative PCR detection system of multi-throughput samples of claim 1, wherein the anti-distortion lens set is composed of two plano-convex lenses and a biconvex lens, which are coaxially disposed, and the biconvex lens is located between the two plano-convex lenses, and the convex surfaces of the two plano-convex lenses are respectively opposite to two sides of the biconvex lens.

8. The real-time fluorescence quantitative PCR detection system of claim 2, wherein the detection light path further includes a plurality of optical filters disposed at the incident end of the imaging device, one of the optical filters corresponds to one of the groups of laser light sources, the optical filter is capable of filtering the laser light emitted from the corresponding laser light source, and the optical filter is capable of transmitting the fluorescence emitted from the sample excited by the corresponding laser light source.

9. The real-time fluorescent quantitative PCR detection system of multi-throughput samples of claim 2, wherein the excitation light path further comprises a rod mirror disposed between the laser light source and the optical fiber.

10. The system for real-time fluorescent quantitative PCR detection of multi-throughput samples of claim 9, wherein the rod mirrors include a first rod mirror and a second rod mirror, both of which are disposed coaxially with the excitation light source; the length direction of the first rod mirror is vertical to the fast axis of the laser; the length direction of the second rod mirror is perpendicular to the slow axis of the laser.