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Detecting Pyronin Y Labeled RNA Transcripts in Live Cell Microenvironments by phasor-FLIM Analysis

Detecting Pyronin Y Labeled RNA Transcripts in Live Cell Microenvironments by phasor-FLIM Analysis

Laura M Andrews, Mark R Jones, Michelle A Digman, Enrico Gratton

Methods Appl Fluoresc. 2013 Mar 1;1(1):015001.

PMID: 24563776

Abstract:

Pyronin Y is an environment-sensitive probe which labels all double-stranded RNA in live cells. Methods to determine which RNA species Pyronin Y may be labeling are limited due to the lack of studies aimed at determining whether this probe has different spectroscopic properties when bound to specific transcripts. A major issue is that transcripts are difficult to isolate and study individually. We detected transcripts directly in their biological environment allowing us to identify RNA species on the basis of their location in the cell. We show that the phasor approach to lifetime analysis has the sensitivity to determine at least six different RNA species in live fibroblast cells. The detected lifetime differences were consistent among cells. To our knowledge this is the first application of a spectroscopic technique aimed at identifying Pyronin Y labeled RNA subtypes in living cells.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP92320	Pyronin Y		92-32-0	Price

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