International Journal of Molecular Medical Science, 2024, Vol.14, No.1, 29-41 http://medscipublisher.com/index.php/ijmms 31 1.2 Single-cell ATAC sequencing (scATAC-seq) The scATAC-seq technique is used to analyze the accessibility of chromatin in individual cells to infer the open status of gene regulatory regions. This is essential for understanding the mechanisms of cell type-specific gene expression regulation. Conklin et al. (2022) proposed that the development of TEA-seq technology is a three-mode sequencing method that can simultaneously measure the transcriptome (scRNA-seq), epitopes, and chromatin accessibility (scATAC-seq) of a single cell, providing a new tool for cell type-specific gene regulation and expression. According to Patruno et al. (2023), the launch of ArchR software package provides a rapid and comprehensive analysis of single-cell chromatin accessibility data, including single-cell clustering, cell type identification, DNA element and gene link analysis, etc., which greatly accelerates the understanding of gene regulation (Granja et al., 2021). Schep et al. (2017) suggest that chromVAR, an R package for analyzing sparse single-cell ATAC-seq data, is able to accurately cluster scATAC-seq profiles and identify known and new moths associated with changes in chromatin accessibility by estimating accessibility gains or losses within peaks that share the same moths or notes. These studies not only expand the understanding of chromatin accessibility and gene regulation mechanisms at the single-cell level, but also provide new tools and analytical frameworks for future research. 1.3 Single cell mass spectrometry (CyTOF) In cell biology research, a technology called CyTOF (Cytometry by Time-Of-Flight), with its unique advantages, is gradually becoming an important tool to study the phenotype and function of immune cells. The core of CyTOF technology is its ability to label proteins on the cell surface and inside the cell, thereby quantifying tens to hundreds of proteins at the single-cell level. This property allows researchers to gain insight into the complexity and diversity of cells in the microscopic world, which in turn reveals the nature of life activities. According to Wang and Navin's (2015) study, CyTOF has higher parametric capacity and lower background noise compared with traditional flow cytometry. Although flow cytometry can also analyze proteins on the cell surface to a certain extent, due to the limitations of its working principle, it is often difficult to analyze multiple proteins at the same time, and the background noise is high, affecting the accuracy of the results. CyTOF technology, through its unique time-flight principle, can significantly increase the parameter capacity of analysis while maintaining high sensitivity, making it possible to analyze tens or even hundreds of proteins at the same time in a single experiment. Proserpio and Mahata (2015) proposed that the application of CyTOF technology is even more convenient in the study of immune cells. As an important force of human body to resist the invasion of foreign pathogens, the diversity of immune cells' phenotypes and functions is crucial for maintaining body homeostasis. CyTOF technology can accurately quantify the expression levels of receptors, ligands and intracellular signaling molecules on the surface of immune cells, thereby revealing the mechanism of action of different immune cell subsets in the occurrence and development of diseases. Researchers can use CyTOF technology to analyze the phenotype and function of immune cells in the tumor microenvironment. By comparing the immune cells in tumor tissue and healthy tissue, researchers can find the changes in the type, number and functional status of tumor-infiltrating immune cells, providing new ideas and methods for tumor immunotherapy. 1.4 Single cell imaging technology Chen et al. (2015) believe that single-cell imaging, as an important tool for modern biological research, covers both optical microscopy and electron microscopy, providing intuitive information on cell morphology, location and interactions. These techniques allow scientists to delve deeper into the complex mechanisms inside cells, opening up new horizons for biomedical research. Optical microscopy is an important part of single cell imaging. By utilizing different wavelengths of light and special fluorescent markers, light microscopes are able to reveal the fine structure and function inside cells. Fluorescence microscopy allows specific protein or nucleic acid molecules to be clearly visible under the microscope by exciting fluorescent molecules within cells. This technique provides
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