title: In vivo photoswitchable flow cytometry for direct tracking of single circulating tumor cells.
abstract: 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.
authors.affiliation: Winthrop P. Rockefeller Cancer Institute, Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences (UAMS), 4301 West Markham, Little Rock, AR 72205, USA.
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Nedosekin
authors.affiliation: Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
authors.forename: Vladislav V
authors.initials: VV
authors.lastname: Verkhusha
authors.affiliation: Moscow Institute of Physics and Technology, 9 Inststitutskii pereulok, Dolgoprudny, Moscow Region 141700, Russian Federation.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Melerzanov
authors.affiliation: Winthrop P. Rockefeller Cancer Institute, Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences (UAMS), 4301 West Markham, Little Rock, AR 72205, USA; Moscow Institute of Physics and Technology, 9 Inststitutskii pereulok, Dolgoprudny, Moscow Region 141700, Russian Federation.
authors.forename: Vladimir P
authors.initials: VP
authors.lastname: Zharov
authors.affiliation: Winthrop P. Rockefeller Cancer Institute, Arkansas Nanomedicine Center, University of Arkansas for Medical Sciences (UAMS), 4301 West Markham, Little Rock, AR 72205, USA. Electronic address: egalanzha@uams.edu.
authors.forename: Ekaterina I
authors.initials: EI
authors.lastname: Galanzha
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D058948:Cell Tracking / Q000379:methods*; D005434:Flow Cytometry; D008164:Luminescent Proteins / Q000737:chemistry*; D051379:Mice; D008819:Mice, Nude; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D055668:Photochemical Processes; D051381:Rats
chemical_list: D008164:Luminescent Proteins