Lysosome-targeting pH indicator based on peri-fused naphthalene monoimide with superior stability for long term live cell imaging.

Journal of Materials Chemistry. B, Materials for Biology and Medicine
Astrid TannertUte Neugebauer

Abstract

Lysosomes, the acidic degradation compartments of eukaryotic cells, play an essential role in many physiological processes. Their dysfunction is associated with a number of diseases, which are often related to an altered localization or luminal pH. Thus, the in-depth characterization of lysosomes within the intact eukaryotic cell is of utmost interest. For microscopic evaluation of lysosomal distribution and acidity, a number of labels have been developed, but many showed poor organelle specificity or rapid clearing from lysosomes, rendering them unsuitable for long-term observations. Here, we describe the synthesis and spectroscopic properties of a novel small molecule marker for lysosomes based on naphthalene monoimide with reversible, pH-dependent spectral shifts in both the absorption and the emission spectrum and acidity-associated changes in fluorescence lifetime. The dye can be excited either with single- or two-photon excitation and appears to be very stably associated with lysosomes for several days. We used this chromophore to detect chemically-induced changes of lysosomal pH in HeLa cells by ratiometric and FLIM imaging.

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Citations

Apr 20, 2021·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Xiaoliang LiWenming Xu
Oct 26, 2021·Chembiochem : a European Journal of Chemical Biology·Pankaj E HandeSantosh J Gharpure

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Methods Mentioned

BETA
transfection
Fluorescence
FCS
PMA

Software Mentioned

Hyperchem
LS
gnuplot
SymphoTime
AMBER99
Gaussian
SPCImage
Fiji
NMI
EasyTau

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