TY - CHAP
T1 - Live cell microscopy of DNA damage response in Saccharomyces cerevisiae
AU - Pinela da Silva, Sonia Cristina
AU - Gallina, Irene
AU - Eckert-Boulet, Nadine Valerie
AU - Lisby, Michael
PY - 2012
Y1 - 2012
N2 - Fluorescence microscopy of the DNA damage response in living cells stands out from many other DNA repair assays by its ability to monitor the response to individual DNA lesions in single cells. This is particularly true in yeast, where the frequency of spontaneous DNA lesions is relatively low compared to organisms with much larger genomes such as mammalian cells. Single cell analysis of individual DNA lesions allows specific events in the DNA damage response to be correlated with cell morphology, cell cycle phase, and other specific characteristics of a particular cell. Moreover, fluorescence live cell imaging allows for multiple cellular markers to be monitored over several hours. This chapter reviews useful fluorescent markers and genotoxic agents for studying the DNA damage response in living cells and provides protocols for live cell imaging, time-lapse microscopy, and for induction of site-specific DNA lesions.
AB - Fluorescence microscopy of the DNA damage response in living cells stands out from many other DNA repair assays by its ability to monitor the response to individual DNA lesions in single cells. This is particularly true in yeast, where the frequency of spontaneous DNA lesions is relatively low compared to organisms with much larger genomes such as mammalian cells. Single cell analysis of individual DNA lesions allows specific events in the DNA damage response to be correlated with cell morphology, cell cycle phase, and other specific characteristics of a particular cell. Moreover, fluorescence live cell imaging allows for multiple cellular markers to be monitored over several hours. This chapter reviews useful fluorescent markers and genotoxic agents for studying the DNA damage response in living cells and provides protocols for live cell imaging, time-lapse microscopy, and for induction of site-specific DNA lesions.
U2 - 10.1007/978-1-61779-998-3_30
DO - 10.1007/978-1-61779-998-3_30
M3 - Book chapter
C2 - 22941621
SN - 978-1-61779-997-6
T3 - Methods in Molecular Biology
SP - 433
EP - 443
BT - DNA repair protocols
PB - Springer Science+Business Media
ER -