Simultaneous visualization of Early- and Late S-phase replicating DNA in the same cell nucleus by fluorescence imaging and FLIM-FRET approach (IMAGE)
Caption
(a) Schematics of approach: cultured cells sequentially labeled with CldU in early S-phase and with IdU in the late S-phase were followed into subsequent cell generations, enabling to visualize segregated chromosome territories with stained early and late S-phase labeled chromatin. Label segregation enables analysis of chromatin compaction within individual chromosome territories. (b-c) Cells were fixed at various intervals following the exposure to halogenated nucleotides and stained (b) for CldU (AlexaFluor 546, green) and (c) IdU (AlexaFluor647, red) as shown in the panel (a). Representative fluorescence intensity (b-e), lifetime images (f) and FRET efficiency (g-h) of the chromosome territories formation process after labeling of the Early and Late S-phase replicating chromatin domains. High FRET areas (red) indicate a close proximity between the chromatin fibers replicated in Early- and Late S-phase.
Credit
by Svitlana M. Levchenko, Artem Pliss, Xiao Peng, Paras N. Prasad and Junle Qu
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Credit must be given to the creator.
License
CC BY