Journal
TALANTA
Volume 269, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2023.125460
Keywords
Single cell heterogeneity; In -situ detection; Real-time isothermal amplification
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Single cell heterogeneity plays an important role in many biological phenomena, and distinguishing cells with specific mutations is beneficial for clinical diagnosis and drug screening. This study introduces a gel plate platform that can distinguish cell types based on their phenotypes, with low equipment requirement. The gel platform has potential in point-of-care circumstances and single-cell stimulation responses that prioritize efficiency and simplicity.
Single cell heterogeneity plays an important role in many biological phenomena and distinguishing cells that exhibit certain mutation in sample could benefit clinical diagnose and drug screening. Typical single cell detection methods such as flow cytometry, in-situ hybridization, real-time amplification or sequencing test either protein or nucleic acid as target and usually require specialized instruments. Joint measurement of the both types of targets could be done by combining the above strategies precisely but also unwieldly. Methods for rapidly and parallelly screening single cells with target genotype and antigen is needed. In this study, we describe a gel plate platform to distinguish cell types based on their phenotypes on target gene and antigen with low equipment requirement. Integrated cell lysis and immobilization were done in the gel solidification step, after which antibody hybridization and real-time amplification were sequentially carried out without losing the original loci information of individual single cells so the three types of information of individual single cells could be combined to distinguished cells with expected genotype and phenotype. The easy-to-use gel platform has potential in point-of-care circumstances and single-cell stimulation response that have high requirements on efficiency and simplicity.
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