Cargando…
Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination
The simultaneous quantification of multi-miRNAs in single cells reveals cellular heterogeneity, and benefits the subtypes discrimination of cancer cells . Though micro-droplet techniques enable successful single cell encapsulation, the isolated and restricted reaction space of microdroplets causes c...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8848760/ https://www.ncbi.nlm.nih.gov/pubmed/35308856 http://dx.doi.org/10.1039/d1sc05304c |
_version_ | 1784652324231184384 |
---|---|
author | Wang, Yingfei Fang, Yanyun Zhu, Yu Bi, Shiyi Liu, Ying Ju, Huangxian |
author_facet | Wang, Yingfei Fang, Yanyun Zhu, Yu Bi, Shiyi Liu, Ying Ju, Huangxian |
author_sort | Wang, Yingfei |
collection | PubMed |
description | The simultaneous quantification of multi-miRNAs in single cells reveals cellular heterogeneity, and benefits the subtypes discrimination of cancer cells . Though micro-droplet techniques enable successful single cell encapsulation, the isolated and restricted reaction space of microdroplets causes cross-reactions and inaccuracy for simultaneous multi-miRNAs quantification. Herein, we develop a hydrogel microbead based strategy for the simultaneous sensitive quantification of miRNA-21, 122 and 222 in single cells. Single cells are encapsulated and undergo cytolysis in hydrogel microbeads. The three target miRNAs are retained in the microbead by pre-immobilized capture probes, and activate rolling circle amplification (RCA) reactions. The RCA products are hybridized with corresponding dye labelled DNA reporters, and the respective fluorescence intensities are recorded for multi-miRNA quantification. The porous structure of the hydrogel microbeads allows the free diffusion of reactants and easy removal of unreacted DNA strands, which effectively avoids nonspecific cross-reactions. Clear differentiation of cellular heterogeneity and subpopulation discrimination are achieved for three kinds of liver cancer cells and one normal liver cell. |
format | Online Article Text |
id | pubmed-8848760 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-88487602022-03-17 Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination Wang, Yingfei Fang, Yanyun Zhu, Yu Bi, Shiyi Liu, Ying Ju, Huangxian Chem Sci Chemistry The simultaneous quantification of multi-miRNAs in single cells reveals cellular heterogeneity, and benefits the subtypes discrimination of cancer cells . Though micro-droplet techniques enable successful single cell encapsulation, the isolated and restricted reaction space of microdroplets causes cross-reactions and inaccuracy for simultaneous multi-miRNAs quantification. Herein, we develop a hydrogel microbead based strategy for the simultaneous sensitive quantification of miRNA-21, 122 and 222 in single cells. Single cells are encapsulated and undergo cytolysis in hydrogel microbeads. The three target miRNAs are retained in the microbead by pre-immobilized capture probes, and activate rolling circle amplification (RCA) reactions. The RCA products are hybridized with corresponding dye labelled DNA reporters, and the respective fluorescence intensities are recorded for multi-miRNA quantification. The porous structure of the hydrogel microbeads allows the free diffusion of reactants and easy removal of unreacted DNA strands, which effectively avoids nonspecific cross-reactions. Clear differentiation of cellular heterogeneity and subpopulation discrimination are achieved for three kinds of liver cancer cells and one normal liver cell. The Royal Society of Chemistry 2022-01-27 /pmc/articles/PMC8848760/ /pubmed/35308856 http://dx.doi.org/10.1039/d1sc05304c Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/ |
spellingShingle | Chemistry Wang, Yingfei Fang, Yanyun Zhu, Yu Bi, Shiyi Liu, Ying Ju, Huangxian Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title | Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title_full | Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title_fullStr | Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title_full_unstemmed | Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title_short | Single cell multi-miRNAs quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
title_sort | single cell multi-mirnas quantification with hydrogel microbeads for liver cancer cell subtypes discrimination |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8848760/ https://www.ncbi.nlm.nih.gov/pubmed/35308856 http://dx.doi.org/10.1039/d1sc05304c |
work_keys_str_mv | AT wangyingfei singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination AT fangyanyun singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination AT zhuyu singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination AT bishiyi singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination AT liuying singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination AT juhuangxian singlecellmultimirnasquantificationwithhydrogelmicrobeadsforlivercancercellsubtypesdiscrimination |