Nanoscale spatial induction of ultraviolet photoproducts in cellular DNA by three-photon near-infrared absorption.
The high-resolution spatial induction of ultraviolet (UV) photoproducts in mammalian cellular DNA is a goal of many scientists who study UV damage and repair. Here we describe how UV photoproducts can be induced in cellular DNA within nanometre dimensions by near-diffraction-limited 750 nm infrared laser radiation. The use of multiphoton excitation to induce highly localized DNA damage in an individual cell nucleus or mitochondrion will provide much greater resolution for studies of DNA repair dynamics and intracellular localization as well as intracellular signalling processes and cell-cell communication. The technique offers an advantage over the masking method for localized irradiation of cells, as the laser radiation can specifically target a single cell and subnuclear structures such as nucleoli, nuclear membranes or any structure that can be labelled and visualized by a fluorescent tag. It also increases the time resolution with which migration of DNA repair proteins to damage sites can be monitored. We define the characteristics of localized DNA damage induction by near-infrared radiation and suggest how it may be used for new biological investigations.
Meldrum
Rosalind A
RA
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK. r.meldrum@bham.ac.uk
Botchway
Stanley W
SW
Wharton
Christopher W
CW
Hirst
Graeme J
GJ
D000818:Animals; D016466:CHO Cells; D002450:Cell Communication; D002467:Cell Nucleus / Q000528:radiation effects*; D002478:Cells, Cultured; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair; D049452:Green Fluorescent Proteins; D007259:Infrared Rays; D007425:Intracellular Membranes / Q000528:radiation effects; D007834:Lasers; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D051381:Rats; D015398:Signal Transduction; D014466:Ultraviolet Rays
D008164:Luminescent Proteins; D018809:Proliferating Cell Nuclear Antigen; D049452:Green Fluorescent Proteins; D004247:DNA