Spatially Encoded Single-Cell Libraries for Accurate Tissue Location
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Solution Overview
Problem
Current single-cell research technologies can only analyze nucleic acid information and some protein information at the single-cell level, but lack the ability to accurately determine the spatial location of individual cells within tissues, leading to distorted cellular localization due to RNA diffusion and multiple cell layers in slices.
Innovation Solution
A method for constructing a single-cell library containing spatial information by using a chip with nucleic acid molecules having known base sequences that bind to tissue cells, followed by dissociation and sequencing to obtain a transcriptome and spatial tag library, enabling simultaneous analysis of genetic and spatial information at the single-cell level.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If in situ capture technology is used to analyze RNA information at tissue location level, then spatial information can be obtained, but the true cellular localization of RNA is distorted due to random diffusion of RNA and presence of multiple cell layers in slices
Solution Approach 1:
The tissue is sectioned into thin slices (5-10 μm thickness) to separate overlapping cell layers, and each slice is processed independently to preserve the spatial information of individual cells without contamination from adjacent layers
Solution Approach 2:
A nucleic acid chip with spatially encoded probes serves as an intermediary substrate that captures RNA molecules directly at their location of origin within the tissue slice, preventing diffusion-related distortion by providing a fixed reference framework
2Measurement precision
If conventional single-cell research technology is used to analyze nucleic acid information, then single-cell level analysis can be achieved, but spatial location information of individual cells within tissues cannot be obtained
Solution Approach 1:
The method merges single-cell dissociation technology with spatially encoded nucleic acid capture by combining steps of tissue sectioning, chip-based probe attachment, cell lysis, and spatially-resolved RNA capture into a unified workflow that preserves both single-cell resolution and spatial location information
Solution Approach 2:
The spatial location information is captured by creating a copy of the spatial coordinates through the nucleic acid chip's encoded probe positions, which record the original location of each cell's RNA molecules before dissociation and processing
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables accurate determination of genetic information and spatial location of single cells within tissues, overcoming the limitations of existing technologies by directly analyzing spatial information without simulation, thus providing a deeper understanding of nucleic acid information at the tissue level.
Implementation Method 1
nucleic acid molecules with a known base sequence... bind to cells of a slice of tissue to be tested
Data Source
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AI summary
Disclosed in the present invention are a method for constructing a single cell library containing tissue location information and a sequencing method. The construction method comprises the following steps: S10, providing a chip, wherein the chip is provided with a plurality of sites, each site is provided with nucleic acid molecules having a known base sequence, and the nucleic acid molecules contain location information; S20, enabling a tissue slice to be tested to be in contact with the chip, so as to bind the nucleic acid molecules having the known base sequence in the chip to cells of said tissue slice to obtain a bound tissue slice; S30, dissociating the bound tissue slice to prepare a single cell suspension; S40, constructing a library for the single cell suspension by means of a high-throughput single cell library construction and sequencing platform, to obtain a transcriptome library and a spatial barcoded library; and S50, sequencing the transcriptome library and the spatial barcoded library to obtain a single cell library containing tissue location information. According to the method, spatial location information of a single cell can be tested while analyzing genetic information on a single cell level.