Multi-Phenotypic subtyping of circulating tumor cells using sequential fluorescent quenching and restaining

Daniel L. Adams, R. Katherine Alpaugh, Susan Tsai, Cha Mei Tang, Steingrimur Stefansson

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


In tissue biopsies formalin fixed paraffin embedded cancer blocks are micro-sectioned producing multiple semi-identical specimens which are analyzed and subtyped proteomically, and genomically, with numerous biomarkers. In blood based biopsies (BBBs), blood is purified for circulating tumor cells (CTCs) and clinical utility is typically limited to cell enumeration, as only 2-3 positive fluorescent markers and 1 negative marker can be used. As such, increasing the number of subtyping biomarkers on each individual CTC could dramatically enhance the clinical utility of BBBs, allowing in depth interrogation of clinically relevant CTCs. We describe a simple and inexpensive method for quenching the specific fluors of fluorescently stained CTCs followed by sequential restaining with additional biomarkers. As proof of principle a CTC panel, immunosuppression panel and stem cell panel were used to sequentially subtype individual fluorescently stained patient CTCs, suggesting a simple and universal technique to analyze multiple clinically applicable immunomarkers from BBBs.

Original languageEnglish (US)
Article number33488
JournalScientific reports
StatePublished - Sep 20 2016

ASJC Scopus subject areas

  • General


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