TY - JOUR
T1 - In vivo photoswitchable flow cytometry for direct tracking of single circulating Tumor Cells
AU - Nedosekin, Dmitry A.
AU - Verkhusha, Vladislav V.
AU - Melerzanov, Alexander V.
AU - Zharov, Vladimir P.
AU - Galanzha, Ekaterina I.
N1 - Funding Information:
We thank L.J. Hennings for her histologic contributions and S. Foster and J. Ye for assistance with cell culturing and imaging. This work was supported by grants CA131164 and EB009230 (both to V.P.Z.), GM073913 and CA164468 (both to V.V.V.) from NIH, grant W81XWH-11-1-0129 from the Department of Defense, and grant UL1TR000039 from the Arkansas Breast Cancer Research Program (both to E.I.G.), and grants from the Arkansas Biosciences Institute and the Translational Research Institute at UAMS (both to V.P.Z.). We also thank the Office of Grants and Scientific Publications at UAMS for editorial assistance.
PY - 2014/6/19
Y1 - 2014/6/19
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84903179491&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84903179491&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2014.03.012
DO - 10.1016/j.chembiol.2014.03.012
M3 - Article
C2 - 24816228
AN - SCOPUS:84903179491
SN - 1074-5521
VL - 21
SP - 792
EP - 801
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 6
ER -