Photoinduced damage resulting from fluorescence imaging of live cells.
The widespread application of fluorescence microscopy to study live cells has led to a greater understanding of numerous biological processes. Many techniques have been developed to uniquely label structures and track metabolic pathways using fluorophores in live cells. However, the photochemistry of nonnative compounds and the deposition of energy into the cell during imaging can result in unexpected and unwanted side effects. Herein, we examine potential live cell damage by first discussing common imaging considerations and modalities in fluorescence microscopy. We then consider several mechanisms by which various photochemical and photophysical phenomena cause cellular damage and introduce techniques that have leveraged these phenomena to intentionally create damage inside cells. Reviewing conditions under which intentional damage occurs can allow one to better predict when unintentional damage may be important. Finally, we delineate ways of checking for and reducing photochemical and photophysical damage.
Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-3290, USA.
Matthew K
MK
Daddysman
Michael A
MA
Tycon
Christopher J
CJ
Fecko
D016477:Artifacts; D002478:Cells, Cultured; D004249:DNA Damage; D008027:Light; D008856:Microscopy, Fluorescence; D010084:Oxidation-Reduction / Q000528:radiation effects; D055668:Photochemical Processes; D059010:Single-Cell Analysis