CN218212647U - Fluorescence excitation detection system and nucleic acid constant-temperature amplification instrument - Google Patents
Fluorescence excitation detection system and nucleic acid constant-temperature amplification instrument Download PDFInfo
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- CN218212647U CN218212647U CN202222623503.7U CN202222623503U CN218212647U CN 218212647 U CN218212647 U CN 218212647U CN 202222623503 U CN202222623503 U CN 202222623503U CN 218212647 U CN218212647 U CN 218212647U
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Abstract
The utility model discloses a fluorescence arouses detecting system and nucleic acid constant temperature amplification appearance, fluorescence arouses detecting system includes more than one target gene light path detection device and the interior mark gene light path detection device of more than one, target gene light path detection device is used for sending the excitation light and receives the fluorescence signal that the target gene sent towards the target gene on the detection consumptive material, interior mark gene light path detection device is used for sending the excitation light and receiving the fluorescence signal that interior mark gene sent towards the interior mark gene on the detection consumptive material, target gene light path detection device and interior mark gene light path detection device interval enclose locate the outside that detects the consumptive material, then arouse detecting system through fluorescence not only can carry out fluorescence detection to the target gene, can also carry out fluorescence detection to interior mark gene, thereby make the testing result of target gene can establish the contrast with the testing result of interior mark gene, in order to reach the purpose that improves the precision of testing result.
Description
Technical Field
The utility model belongs to the technical field of optical detection, concretely relates to fluorescence excitation detecting system and nucleic acid constant temperature amplification appearance.
Background
The isothermal amplification technology rapidly developed in recent years gradually replaces the traditional PCR (Polymerase Chain Reaction) amplification technology, is simpler and more convenient than the traditional PCR amplification technology in technical and equipment requirements, does not need multiple temperature change processes, is rapid and accurate in target detection, does not need high-end equipment, and has wide application prospects in clinical and field detection. However, in both the conventional PCR amplification and isothermal amplification detection technologies, the amount of nucleic acid amplification needs to be detected by fluorescence indication, but the existing detection device has low accuracy of detection result due to lack of target control in the fluorescence detection process.
SUMMERY OF THE UTILITY MODEL
To the not enough of the aforesaid current, the utility model provides a fluorescence arouses detecting system and nucleic acid constant temperature amplification appearance aims at solving current check out test set and lacks the target contrast in fluorescence detection process and lead to detecting the technical problem that the precision of structure is low on the one side.
In order to achieve the above object, the present invention provides a fluorescence excitation detection system for performing fluorescence detection on detection consumables, wherein the fluorescence excitation detection system comprises more than one target gene light path detection device and more than one internal standard gene light path detection device; the target gene light path detection device is used for emitting exciting light towards a target gene on the detection consumable and receiving a fluorescent signal emitted by the target gene; the internal standard gene light path detection device is used for emitting excitation light towards the internal standard gene on the detection consumable and receiving a fluorescence signal emitted by the internal standard gene; the target gene light path detection device and the internal standard gene light path detection device are arranged outside the detection consumable at intervals.
The embodiment of the utility model provides an in, target gene light path detection device's quantity is two, and interior mark gene light path detection device's quantity is one, and two target gene light path detection device are used for one-to-one ground to send excitation light and correspond respectively and receive the fluorescence signal that two target genes sent towards two target genes on the detection consumptive material to interior mark gene light path detection device and two target gene light path detection device evenly spaced enclose the outside of locating the detection consumptive material.
The embodiment of the utility model provides an in, target gene light path detection device and interior mark gene light path detection device all include: a fluorescence excitation component and a fluorescence receiving component; the fluorescence excitation assembly comprises a bullet light source piece and a first optical filter, wherein the bullet light source piece emits light towards the first optical filter and irradiates the liquid to be detected of the detection consumable so as to excite the liquid to be detected to emit fluorescence; the fluorescence receiving assembly receives fluorescence emitted by the liquid to be detected and converts a fluorescence signal into an electric signal.
In the embodiment of the present invention, the included angle between the fluorescence excitation light path formed by the fluorescence excitation component and the fluorescence receiving light path formed by the fluorescence receiving component is 75 ° to 105 °.
The embodiment of the utility model provides an in, fluorescence receiving element is including collimating lens, second light filter, focusing lens and the photodiode that sets up in proper order at the interval, and collimating lens is used for receiving the fluorescence that waits to detect the liquid and send to make fluorescence pass through second light filter and focusing lens directive photodiode in proper order.
In the embodiment of the present invention, the number of the focusing lenses is two, and two focusing lens intervals are disposed between the second optical filter and the photodiode.
In an embodiment of the present invention, the collimating lens is a plano-convex lens, and the focusing lens is a biconvex lens.
In an embodiment of the present invention, the divergence angle of the bullet light source is less than 20 °.
The embodiment of the utility model provides an in, fluorescence arouses detecting system still includes controlling means, and controlling means arouses the bullet head light source spare communication connection in the subassembly with every fluorescence respectively to can control and carry out the timesharing and light.
In order to achieve the above object, the present invention also provides a nucleic acid constant temperature amplification apparatus, wherein the nucleic acid constant temperature amplification apparatus comprises a fluorescence excitation detection system according to the above.
Through the technical scheme, the embodiment of the utility model provides a fluorescence excitation detecting system has following beneficial effect:
when the fluorescence excitation detection system is used, the target gene light path detection device can emit excitation light towards the target gene on the detection consumable and receive a fluorescence signal emitted by the target gene, the internal standard gene light path detection device can emit excitation light towards the internal standard gene on the detection consumable and receive a fluorescence signal emitted by the internal standard gene, and then the fluorescence excitation detection system can be used for performing fluorescence detection on the target gene and the internal standard gene, so that the detection result of the target gene can be contrasted with the detection result of the internal standard gene, and the purpose of improving the precision of the detection result is achieved.
Other features and advantages of the present invention will be described in detail in the detailed description which follows.
Drawings
The accompanying drawings, which are included to provide an understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention, but do not constitute a limitation of the invention. In the drawings:
FIG. 1 is a schematic view of a partial structure of a nucleic acid constant temperature amplification apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a fluorescence excitation detection system according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of a fluorescence receiving assembly according to an embodiment of the present invention.
Description of the reference numerals
100. 200 internal standard gene light path detection device of target gene light path detection device
1. Fool-proof structure of detection consumable 11
2. Fluorescence excitation assembly 21 bullet head light source part
22. First optical filter 3 fluorescence receiving assembly
31. Second filter of collimating lens 32
33. Focusing lens 34 photodiode
Detailed Description
The following detailed description of the embodiments of the present invention will be made with reference to the accompanying drawings. It is to be understood that the specific embodiments described herein are for purposes of illustration and explanation only and are not intended to limit the present invention.
The fluorescence excitation detection system of the present invention is described below with reference to the drawings.
As shown in fig. 1, the utility model provides a fluorescence excitation detecting system, fluorescence excitation detecting system is used for carrying out fluorescence detection to detecting consumptive material 1, and wherein, fluorescence excitation detecting system includes:
more than one target gene light path detection device 100, used for emitting excitation light towards the target gene on the detection consumable 1 and receiving the target gene emitted fluorescence signal;
more than one internal standard gene light path detection device 200 for emitting excitation light towards the internal standard gene on the detection consumable 1 and receiving a fluorescence signal emitted by the internal standard gene;
wherein, the target gene light path detection device 100 and the internal standard gene light path detection device 200 are surrounded on the outer side of the detection consumable 1 at intervals.
When the fluorescence excitation detection system is used, the target gene light path detection device 100 can emit excitation light towards the target gene on the detection consumable 1 and receive a fluorescence signal emitted by the target gene, the internal standard gene light path detection device 200 can emit excitation light towards the internal standard gene on the detection consumable 1 and receive a fluorescence signal emitted by the internal standard gene, so that the fluorescence excitation detection system can perform fluorescence detection on the target gene and the internal standard gene, and the detection result of the target gene can be compared with the detection result of the internal standard gene, so that the purpose of improving the precision of the detection result is achieved.
It should be especially noted that, the detection consumable 1 is formed with the first amplification reaction chamber and the second amplification reaction chamber for waiting to detect the liquid holding, and first amplification reaction chamber is used for holding the liquid of waiting to detect of target gene, and the second amplification reaction chamber is used for holding the liquid of waiting to detect of interior label gene, and then target gene light path detection device 100 corresponds first amplification reaction chamber and encloses the outside of detecting consumable 1, and interior label gene light path detection device 200 corresponds second amplification reaction chamber and encloses the outside of detecting consumable 1.
The embodiment of the utility model provides an in, the quantity of target gene light path detection device 100 can be two, and interior mark gene light path detection device 200's quantity is one, and two target gene light path detection device 100 are used for one-to-one ground to send excitation light and correspond respectively and receive the fluorescence signal that two target genes sent towards two target genes on the detection consumptive material 1 to interior mark gene light path detection device 200 and two target gene light path detection device 100 evenly separate and enclose the outside of locating the detection consumptive material 1. That is, the outer side of the detection consumable 1 may be formed with three sets of fluorescence excitation detection optical paths, wherein two sets of fluorescence excitation detection optical paths are formed by two target gene optical path detection devices 100 for performing fluorescence detection on two target genes on the detection consumable 1 respectively, and another set of fluorescence excitation detection optical path is formed by an internal standard gene optical path detection device 200 for performing fluorescence detection on an internal standard gene on the detection consumable 1. The target gene optical path detection device 100 is additionally arranged, so that the aim of enriching detection items can be fulfilled.
Specifically, the number of the first amplification reaction chambers on the detection consumable 1 may be two, so that two target gene optical path detection devices 100 are enclosed at the outer side of the detection consumable 1 in one-to-one correspondence to the two first amplification reaction chambers, and the number of the second amplification reaction chambers on the detection consumable 1 may be one, so that one internal standard gene optical path detection device 200 is enclosed at the outer side of the detection consumable 1 in correspondence to the second amplification reaction chambers.
Referring to fig. 1 and 2, in the embodiment of the present invention, the target gene optical path detection apparatus 100 and the internal standard gene optical path detection apparatus 200 each include: a fluorescence excitation assembly 2 and a fluorescence receiving assembly 3; the fluorescence excitation assembly 2 comprises a bullet light source part 21 and a first optical filter 22, the bullet light source part 21 emits light towards the first optical filter 22, the light irradiates the liquid to be detected in one amplification reaction cavity of the detection consumable 1 through the transmission light of the first optical filter 22, and the liquid to be detected is excited to emit fluorescence; the fluorescence receiving component 3 receives fluorescence emitted by the liquid to be detected and converts a fluorescence signal into an electric signal. The excitation light source of the fluorescence excitation component 2 can adopt a bullet light source part 21, and since the bullet light source part 21 has a focusing function, a focusing lens in the fluorescence excitation component 2 in the prior art can be omitted by arranging the bullet light source part 21, so that the purposes of reducing cost and size are achieved, and the fluorescence excitation component 2 is particularly suitable for household use.
Specifically, when the number of the target gene light path detection devices 100 is two and the number of the internal standard gene light path detection devices 200 is one, the number of the fluorescence excitation components 2 and the number of the fluorescence receiving components 3 are three, the three fluorescence excitation components 2 and the three fluorescence receiving components 3 are combined in pairs to form three groups of fluorescence excitation detection light paths, and the three groups of fluorescence excitation detection light paths are respectively arranged in one-to-one correspondence with the three amplification reaction cavities on the detection consumable 1. Two of the three amplification reaction cavities are first amplification reaction cavities corresponding to the target gene, the other one is a second amplification reaction cavity corresponding to the internal standard gene, and the target gene light path detection device 100 and the internal standard gene light path detection device 200 need to be correspondingly arranged in a fluorescence excitation detection light path according to the first amplification reaction cavity and the second amplification reaction cavity. In addition, in order to facilitate distinguishing the second amplification reaction cavity corresponding to the internal standard gene detection, the detection consumable 1 may be provided with a fool-proof structure 11 at a position corresponding to the second amplification reaction cavity so as to indicate the detection of the internal standard gene. Further, the taper part of the detection consumable 1 between the fluorescence excitation module 2 and the fluorescence receiving module 3 for target gene detection is chamfered correspondingly, while the taper part of the detection consumable 1 between the fluorescence excitation module 2 and the fluorescence receiving module 3 for internal standard gene detection is not chamfered and arrow marks are formed at the corresponding taper parts.
In other embodiments, the fluorescence excitation detection system may also be composed of a plurality of internal standard gene optical path detection devices 200 and a plurality of target gene optical path detection devices 100, wherein the number of the target gene optical path detection devices 100 is an integral multiple of the internal standard gene optical path detection devices 200, and then fluorescence detection may be performed on a plurality of samples in one fluorescence excitation detection system, and the fluorescence detection may be performed synchronously or asynchronously.
The embodiment of the utility model provides an in, the fluorescence that fluorescence arouses subassembly 2 to form arouses the contained angle of the fluorescence receiving light path that light path and fluorescence receiving component 3 formed can be 75 ~ 105, when the quantity of amplification reaction chamber is more than two, detect that 1 outside of consumptive material needs to enclose and is equipped with fluorescence arouse subassembly 2 more than two and fluorescence receiving component 3 more than two, the contained angle that will correspond fluorescence arouse light path and fluorescence receiving light path that an amplification reaction chamber set up sets up to 75 ~ 105, then be convenient for detect the overall arrangement design in the 1 outside of consumptive material. It should be noted that the fluorescence excitation optical path of the fluorescence excitation assembly 2 refers to the central axis of the arrangement of the bullet light source member 21 and the first filter 22, the fluorescence receiving optical path of the fluorescence receiving assembly 3 refers to the central axis of the collimating lens 31, the second filter 32, the focusing lens 33 and the photodiode 34, and the fluorescence excitation optical path formed by the fluorescence excitation assembly 2 and the fluorescence receiving optical path formed by the fluorescence receiving assembly 3 may constitute a fluorescence detection optical path. More specifically, the angle between the fluorescence excitation light path and the fluorescence reception light path may preferably be 90 °.
Referring to fig. 2 and fig. 3, in the embodiment of the present invention, the fluorescence receiving assembly 3 includes a collimating lens 31, a second optical filter 32, a focusing lens 33 and a photodiode 34, which are sequentially disposed at intervals, the collimating lens 31 is used for receiving fluorescence emitted by the liquid to be detected, so that the fluorescence sequentially passes through the second optical filter 32 and the focusing lens 33, and finally emits to the photodiode 34. That is, the collimating lens 31 is disposed close to the solution to be detected in the amplification reaction chamber of the detection consumable 1, and the fluorescence emitted from the solution to be detected first irradiates the collimating lens 31, and then enters the photodiode 34 after sequentially passing through the second optical filter 32 and the focusing lens 33, so as to convert the fluorescence signal into an electrical signal.
In the embodiment of the present invention, the number of the focusing lenses 33 may be two, and two focusing lenses 33 are disposed between the second optical filter 32 and the photodiode 34 at intervals to enhance the focusing effect.
In the embodiment of the present invention, the collimating lens 31 may be a plano-convex lens, and the focusing lens 33 may be a biconvex lens. The collimator lens 31 and the focus lens 33 may be made of glass or plastic, such as OKP-1 or K26R, and the collimator lens 31 may be made of glass and the focus lens 33 may be made of plastic. In addition, the second filter 32 can be a band pass filter, the center wavelength of the second filter 32 is near 520nm, the bandwidth is 20nm, and the cut-off depth outside the pass band is above OD6, so that the part of the excitation light reflected or scattered from the amplification reaction chamber can be effectively blocked, and only the fluorescent component with the wavelength of about 520nm is allowed to pass through; the photodiode 34 may be a silicon photodiode 34, and is preferably small in dark current and large in light-sensing area.
In embodiment 1, the basic parameters of each lens of the fluorescence receiving element 3 along the direction from the liquid to be measured to the photodiode 34 are shown in table 1, with the unit: and (4) millimeter.
TABLE 1 basic parameters of respective mirror surfaces of fluorescence receiving element (example 1)
In table 1, the collimating lens 31 and the focusing lens 33 are made of glass, and the number of the focusing lenses 33 is 2; the surfaces 2 and 3 correspond to the plane and convex surface of the collimating lens 31, respectively, and the rest are similar.
In embodiment 2, the basic parameters of each lens of the fluorescence receiving member 3 are shown in table 2, with the units: and (4) millimeter.
TABLE 2 basic parameters of the respective mirror surfaces of the fluorescence receiving element (example 2)
In table 2, the collimator lens 31 and the focusing lens 33 are made of plastic, and the number of the focusing lenses 33 is 1.
In embodiment 3, the basic parameters of each lens of the fluorescence receiving member 3 are as shown in table 3, and the unit: and (4) millimeter.
TABLE 3 basic parameters of the respective mirror surfaces of the fluorescence receiving element (example 3)
In table 3, the collimator lens 31 is made of glass, the focusing lens 33 is made of plastic, and the number of focusing lenses 33 is 1.
In the embodiment of the present invention, the divergence angle of the bullet light source 21 is smaller than 20 ° to ensure a good focusing action. Specifically, the bullet light source member 21 may be a bullet LED, which may be a type of a 0.5W F8 bullet monochromatic LED, with a radius of curvature at the apex position of about 3.25mm, and a straight line, with a peak wavelength of about 470nm, and a practically used current of about 60 mA. Furthermore, the first filter 22 may preferably be a band pass filter, or a short wavelength pass filter, with an out-of-band cut-off depth of OD4 or more.
The embodiment of the utility model provides an in, fluorescence arouses detecting system still includes controlling means, controlling means arouses the bullet head light source 21 communication connection in the subassembly 2 with every fluorescence respectively to can control and light in timesharing. The correspondingly arranged fluorescence excitation component 2 and the fluorescence receiving component 3 form a group of fluorescence detection paths, and in order to avoid that light rays emitted by the bullet light source part 21 in one group of fluorescence detection paths and fluorescence emitted by the liquid to be detected interfere with the other group of fluorescence detection paths, the control unit can control the bullet light source part 21 in each fluorescence excitation component 2 to be turned on in a time-sharing manner.
In addition, the fluorescence detection method of the fluorescence excitation detection system comprises the following steps:
controlling the light source element in the target gene light path detection device 100 to emit exciting light toward the target gene on the detection consumable 1, so that the photodiode 34 in the target gene light path detection device 100 receives the fluorescent signal emitted by the target gene;
the light source element in the internal standard gene light path detection device 200 is controlled to emit excitation light towards the internal standard gene on the detection consumable 1, so that the photodiode 34 in the internal standard gene light path detection device 200 receives a fluorescence signal emitted by the internal standard gene.
Additionally, the utility model also provides a nucleic acid constant temperature amplification appearance, wherein, nucleic acid constant temperature amplification appearance includes according to above fluorescence arouses detecting system. Since the nucleic acid isothermal amplification instrument adopts all the technical schemes of the above embodiments, at least all the beneficial effects brought by the technical schemes of the above embodiments are achieved, and details are not repeated herein.
In the description of the present invention, it is to be understood that the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any 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 the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," and "fixed" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally formed; may be mechanically coupled, may be electrically coupled or may be in communication with each other; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art without departing from the scope of the present invention.
Claims (10)
1. Fluorescence excitation detection system for fluorescence detection of a detection consumable (1), characterized in that it comprises:
more than one target gene light path detection device (100) which is used for emitting excitation light towards the target gene on the detection consumable (1) and receiving a fluorescent signal emitted by the target gene;
more than one internal standard gene light path detection device (200) is used for emitting excitation light towards the internal standard genes on the detection consumable (1) and receiving fluorescence signals emitted by the internal standard genes;
wherein the target gene light path detection device (100) and the internal standard gene light path detection device (200) are arranged outside the detection consumable (1) in a surrounding mode at intervals.
2. The fluorescence excitation detecting system according to claim 1, wherein the number of the target gene optical path detecting devices (100) is two, the number of the internal standard gene optical path detecting devices (200) is one, the two target gene optical path detecting devices (100) are configured to emit excitation light toward two target genes on the detection consumable (1) in a one-to-one correspondence manner and respectively receive fluorescence signals emitted by the two target genes, and the internal standard gene optical path detecting devices (200) and the two target gene optical path detecting devices (100) are uniformly surrounded at intervals outside the detection consumable (1).
3. The fluorescence excitation detecting system according to claim 1, wherein the target gene optical path detecting device (100) and the internal standard gene optical path detecting device (200) each comprise:
the fluorescence excitation assembly (2) comprises a bullet light source part (21) and a first optical filter (22), wherein the bullet light source part (21) emits light towards the first optical filter (22) and irradiates the liquid to be detected of the detection consumable (1) so as to excite the liquid to be detected to emit fluorescence;
and the fluorescence receiving component (3) is used for receiving a fluorescence signal emitted by the liquid to be detected and converting the fluorescence into an electric signal.
4. The fluorescence excitation detection system according to claim 3, wherein the included angle between the fluorescence excitation light path formed by the fluorescence excitation component (2) and the fluorescence receiving light path formed by the fluorescence receiving component (3) is 75 ° to 105 °.
5. The fluorescence excitation detection system according to claim 3, wherein the fluorescence receiving assembly (3) comprises a collimating lens (31), a second optical filter (32), a focusing lens (33) and a photodiode (34) which are sequentially arranged at intervals, and the collimating lens (31) is configured to receive the fluorescence emitted by the liquid to be detected, so that the fluorescence sequentially passes through the second optical filter (32) and the focusing lens (33) and is emitted to the photodiode (34).
6. The fluorescence excitation detection system according to claim 5, wherein the number of the focusing lenses (33) is two, and two focusing lenses (33) are disposed at intervals between the second filter (32) and the photodiode (34).
7. The fluorescence excitation detection system according to claim 5, wherein the collimator lens (31) is a plano-convex lens and the focusing lens (33) is a biconvex lens.
8. Fluorescence excitation detection system according to claim 3, characterized in that the divergence angle of the bullet light source element (21) is less than 20 °.
9. The fluorescence excitation detection system according to claim 3, further comprising a control device, wherein the control device is respectively connected with the bullet head light source part (21) in each fluorescence excitation assembly (2) in a communication manner so as to control the time-sharing illumination.
10. A nucleic acid isothermal amplification apparatus comprising the fluorescence excitation detection system according to any one of claims 1 to 8.
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