Assessment of RNA flow cytometry probe multiplexing.

Abstract

<p>(<b>a</b>) Flow cytometry plots from three-probe (bcr, abl, 18 s) RNA analysis in the K562 cell line: 18 s rRNA (left) and bcr vs. abl RNA in the 18 s+ events (right). (<b>b</b>) 60x pseudocolored images of sorted bcr+abl+18 s+ cells from K562 cells; Alexa Fluor® 647 labeled bcr (red), Alexa Fluor® 546 labeled abl (green). The bcr/abl fusion transcripts are shown in yellow due to the merging of both the Alexa Fluor® 546 and Alexa Fluor® 647 dyes. Cells were counterstained with DAPI. (<b>c</b>) Graph showing the effects of RNA staining by multiple probes (bcr+abl+18 s) compared to a single target probe on MFIs in RNA flow cytometry. Bars represent the standard deviation for duplicate samples run.</p