Real-time fluorescent quantitation PCR (polymerase chain reaction) instrument

Abstract

The application provides a real-time fluorescent quantitative PCR instrument, including an optical read head, which is composed of at least two optical detection channels, wherein each optical detection channel includes: an excitation light source, a dichroic mirror, an emission light lens, and the detection device; after the excitation light of the excitation light source directly hits the dichroic mirror, the dichroic mirror converts the direction of the light path and shoots vertically to the target tube to be detected EP tube, and the light reflected from the target tube to be detected EP tube is sequentially After passing through a dichroic mirror and a light-emitting lens, it enters the detection device for detection; wherein, each optical detection channel shares the light-emitting lens and the detection device; the light-emitting lens includes a Fresnel lens; each optical channel of the present application The efficiency of collecting light is close, which effectively solves the spherical aberration of the convex lens; the same detection device eliminates the individual differences between devices; the position setting of the optical path effectively avoids the cross interference between the optical paths.

Description

technical field [0001] The invention relates to the technical field of optical detection, in particular to a real-time fluorescence quantitative PCR instrument. Background technique [0002] Polymerase chain reaction (PCR), as a common analysis method in molecular biology, has been widely used in scientific research and clinical fields. Most of the follow-up PCR uses electrophoresis for qualitative and semi-quantitative analysis. Due to the impossibility of quantitative analysis and possible contamination caused by opening the cover, a real-time fluorescent quantitative PCR system came into being. At present, most of the real-time fluorescent quantitative PCR instruments on the market have multi-color fluorescent channels, which can perform multiple PCR and allele, SNP and other analysis at the same time, and the required detection results can be obtained in one experiment. A PCR instrument using multi-color detection generally includes multiple detection optical paths, and...

AI Technical Summary

Problems solved by technology

Generally, there are several ways to realize this structure. One is that the lens and detection structure at the receiving end are separated for each channel, so that different detection structures will have differences between devices, and will increase material costs.

The second is to use the same excitation light source or detection device. Due to the multiplexing of the device, there will be no device difference, but the design of the optical lens and structure is prone to optical path difference and cross interference between optical paths. Specifically, due to the use of optical filters Rotary wheel gating structure, which is often the way of imaging. Due to the difference in optical path difference between the 96-well and the excitation light source and detection device, the excitation and reception efficiency are also different, which will affect the detection results. The collection lens often adopts a plano-convex lens, so due to the different distances and positions of multiple optical paths from the plano-convex lens, it will also cause differences and phase differences in excitation light intensity and collection efficiency; In 500-700nm, integrating 5-6 optical channels in this range will inevitably cause the intersection of the fluorescence collection filter spectrum of the low-band optical channel and the excitation filter spectrum of the adjacent high-band optical channel. phenomenon, see figure 1 , the place marked by the arr

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