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2017 ; 8
(1
): 577
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SRpHi ratiometric pH biosensors for super-resolution microscopy
#MMPMID28924139
Richardson DS
; Gregor C
; Winter FR
; Urban NT
; Sahl SJ
; Willig KI
; Hell SW
Nat Commun
2017[Sep]; 8
(1
): 577
PMID28924139
show ga
Fluorescence-based biosensors have become essential tools for modern biology,
allowing real-time monitoring of biological processes within living cells.
Intracellular fluorescent pH probes comprise one of the most widely used families
of biosensors in microscopy. One key application of pH probes has been to monitor
the acidification of vesicles during endocytosis, an essential function that aids
in cargo sorting and degradation. Prior to the development of super-resolution
fluorescence microscopy (nanoscopy), investigation of endosomal dynamics in live
cells remained difficult as these structures lie at or below the ~250?nm
diffraction limit of light microscopy. Therefore, to aid in investigations of pH
dynamics during endocytosis at the nanoscale, we have specifically designed a
family of ratiometric endosomal pH probes for use in live-cell STED
nanoscopy.Ratiometric fluorescent pH probes are useful tools to monitor
acidification of vesicles during endocytosis, but the size of vesicles is below
the diffraction limit. Here the authors develop a family of ratiometric pH
sensors for use in STED super-resolution microscopy, and optimize their delivery
to endosomes.
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