Single-Cell Spatial Barcode Library for True Tissue Localization
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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 directly determine the spatial location of individual cells within tissues, leading to distorted cellular localization due to RNA diffusion and multiple cell layers.
Innovation Solution
A method for constructing a single-cell library that includes providing a chip with nucleic acid molecules containing spatial information, binding these molecules to tissue slices, dissociating the bound tissue to prepare a single-cell suspension, and using a high-throughput platform to construct and sequence a transcriptome and spatial tag library, thereby enabling simultaneous analysis of genetic and spatial information at the single-cell level.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
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 severely distorted due to random diffusion of RNA and presence of multiple cell layers
Solution Approach 1:
The tissue is sectioned into thin slices (5-20 μm thickness) to separate multiple cell layers, allowing individual cell analysis while preserving spatial context. Each slice contains cells from a specific depth, enabling precise localization without the distortion caused by overlapping layers in thick sections.
Solution Approach 2:
A spatial barcode library is introduced as an intermediary system. Unique spatial barcodes are assigned to specific locations on the chip, and these barcodes are captured along with RNA molecules. The barcodes serve as mediators that link RNA molecules to their original spatial positions, enabling accurate reconstruction of cellular localization despite RNA diffusion.
2Quantity of substance
If traditional single-cell research technology is used to analyze nucleic acid information, then single-cell level genetic information can be obtained, but spatial location information of individual cells within tissues is lost
Solution Approach 1:
The method merges single-cell RNA sequencing with spatial transcriptomics by combining two information streams: genetic information (RNA molecules) and spatial information (location barcodes). Both types of information are captured simultaneously in the same experimental workflow, allowing analysis of nucleic acid composition at single-cell resolution while retaining precise spatial location data.
Solution Approach 2:
The invention adds a spatial dimension to traditional single-cell sequencing. By introducing spatial barcodes that encode positional information and combining them with genetic data, the method transforms a one-dimensional genetic analysis into a two-dimensional analysis that includes both genetic composition and spatial location, enabling mapping of gene expression patterns across tissue architecture.
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 analysis of genetic information and spatial location of single cells within tissues, overcoming the distortion issues of previous methods by directly capturing spatial information at the true physical level of single cells.
Implementation Method 1
nucleic acid molecules with known base sequences in the chip bind to cells of the slice of the tissue to be tested
Data Source
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.