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• W3124903521 abstract "Abstract Recently, advances in fluorescent in-situ hybridization techniques and in imaging technology have enabled visualization and counting of individual RNA molecules in single cells. This has greatly enhanced the resolution in our understanding of transcriptional processes. Here, we adapt a recently published smiFISH protocol (single-molecule inexpensive fluorescent in-situ hybridization) to whole embryos across a range of arthropod model species, and also to non-embryonic tissues. Using multiple fluorophores with distinct spectra and white light laser confocal imaging, we simultaneously detect and separate single RNAs from up to eight different genes in a whole embryo. We also combine smiFISH with cell membrane immunofluorescence, and present an imaging and analysis pipeline for 3D cell segmentation and single-cell RNA counting in whole blastoderm embryos. Finally, using whole embryo single-cell RNA count data, we propose two alternative single-cell variability measures to the commonly used Fano factor, and compare the capacity of these three measures to address different aspects of single-cell expression variability." @default.
• W3124903521 created "2021-02-01" @default.
• W3124903521 creator A5046920134 @default.
• W3124903521 creator A5081319854 @default.
• W3124903521 creator A5087979171 @default.
• W3124903521 date "2021-03-19" @default.
• W3124903521 modified "2023-10-01" @default.
• W3124903521 title "smiFISH and embryo segmentation for single-cell multi-gene RNA quantification in arthropods" @default.
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• W3124903521 doi "https://doi.org/10.1038/s42003-021-01803-0" @default.
• W3124903521 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7979837" @default.
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• W3124903521 hasPublicationYear "2021" @default.
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