In vivo photoswitchable flow cytometry for direct tracking of single circulating Tumor Cells

Dmitry A. Nedosekin, Vladislav V. Verkhusha, Alexander V. Melerzanov, Vladimir P. Zharov, Ekaterina I. Galanzha

Research output: Contribution to journalArticlepeer-review

41 Scopus citations


Photoswitchable fluorescent proteins (PSFPs) that change their color in response to light have led to breakthroughs in studying static cells. However, using PSFPs to study cells in dynamic conditions is challenging. Here we introduce a method for in vivo ultrafast photoswitching of PSFPs that provides labeling and tracking of single circulating cells. Using in vivo multicolor flow cytometry, this method demonstrated the capability for studying recirculation, migration, and distribution of circulating tumor cells (CTCs) during metastasis progression. In tumor-bearing mice, it enabled monitoring of real-time dynamics of CTCs released from primary tumor, identifying dormant cells, and imaging of CTCs colonizing a primary tumor (self-seeding) or existing metastasis (reseeding). Integration of genetically encoded PSFPs, fast photoswitching, flow cytometry, and imaging makes in vivo single cell analysis in the circulation feasible to provide insights into the behavior of CTCs and potentially immune-related and bacterial cells in circulation.

Original languageEnglish (US)
Pages (from-to)792-801
Number of pages10
JournalChemistry and Biology
Issue number6
StatePublished - Jun 19 2014

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry


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