High-density signal decomposition sparse interpretation method applied to space omics and application of high-density signal decomposition sparse interpretation method

Abstract

The invention discloses a high-density signal decomposition sparse interpretation method applied to space omics and application thereof, and the method comprises the steps: dividing all target molecules into anchoring molecules and N groups of sparse molecules, and reading all anchoring molecules and one group of sparse molecules in a connection sequencing mode in each round until the N groups of sparse molecules are completely read. Through N rounds of imaging, high-density target molecule signals which cannot be interpreted are effectively thinned into N groups of low-density signals, then the interpreted N groups of sparse molecules are integrated through anchoring molecule registration, and high-flux and high-density spatial distribution mode information is determined. According to the method, the problems of optical congestion and interference of fluorescence signal points are solved by increasing the number of imaging rounds, splitting, sparse high-density images and exchanging spatial positions, the spatial positions of the high-density signal points are obtained, precise spatial resolution analysis of high-flux and high-density fluorescence signals at the single cell level does not need to be achieved through a super-resolution microscopic imaging system, and the method is simple and convenient to operate. And the current technical limitation is broken.

Description

technical field [0001] The invention relates to the field of high-throughput in situ sequencing, in particular to a method for decomposing and sparsely interpreting high-density signals in space omics and its application. Background technique [0002] Cells are the basic unit of tissue structure and function in living organisms. The traditional high-throughput sequencing method is to read the genetic information of a large number of cells to obtain the "overall" representation of this group of cells. With the deepening of research, people found that there is heterogeneity among cells, and traditional sequencing methods have been unable to mine such "details". Until the emergence of single-cell sequencing technology, it was possible to analyze life activities at the molecular level from the precision of a single cell, and to close-up a single cell, which greatly promoted the development of research fields such as development, cancer, nerves, and immunity. However, during th...

AI Technical Summary

Problems solved by technology

The probe preparation based on single-molecule hybridization is cumbersome and the cost of a large number of fluorescent reading probes is high; the operation steps are complicated, requiring multiple cycles of hybridization-imaging-probe stripping, which requires high system stability; due to the lack of "locating points" ", optical distortion makes the registration of multiple rounds of signals difficult; super-resolution fluorescence microscopy is required, and the requirements for instruments are extremely high

The method based on in situ sequencing hybridization uses rolling circle amplification to amplify the signal. The volume of a single signal spot is larger than that of single-molecule hybridization. Due to the existence of diffraction limit, the detection throughput is limited.

Each cell has tens of thousands to hundreds of thousands of RNAs, and it is extremely difficult to globally analyze the transcriptional profile of the entire cell at one time; because of the existence of the diffraction limit, it will lead to optical congestion and require extremely high optical resolution of the instrument. Therefore, high-precision in situ imaging at low optical resolution has been a major challenge in single-cell biology.

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