Fluorogenic Rhodamine Probes with Pyrrole Substitution Enables STED and Lifetime Imaging of Lysosomes in Live Cells.
Ying ZhouQiuping WangSupphachok ChanmungkalakulXia WuHui XiaoRong MiaoXiaogang LiuYu FangPublished in: Chemistry (Weinheim an der Bergstrasse, Germany) (2024)
Fluorogenic dyes with high brightness, large turn-on ratios, excellent photostability, favorable specificity, low cytotoxicity, and high membrane permeability are essential for high-resolution fluorescence imaging in live cells. In this study, we endowed these desirable properties to a rhodamine derivative by simply replacing the N, N-diethyl group with a pyrrole substituent. The resulting dye, Rh-NH, exhibited doubled Stokes shifts (54 nm) and a red-shift of more than 50 nm in fluorescence spectra compared to Rhodamine B. Rh-NH preferentially exists in a non-emissive but highly permeable spirolactone form. Upon binding to lysosomes, the collective effects of low pH, low polarity, and high viscosity endow Rh-NH with significant fluorescence turn-on, making it a suitable candidate for wash-free, high-contrast lysosome tracking. Consequently, Rh-NH enabled us to successfully explore stimulated emission depletion (STED) super-resolution imaging of lysosome dynamics, as well as fluorescence lifetime imaging of lysosomes in live cells.
Keyphrases
- fluorescent probe
- high resolution
- fluorescence imaging
- living cells
- induced apoptosis
- cell cycle arrest
- photodynamic therapy
- single molecule
- endoplasmic reticulum stress
- magnetic resonance
- endothelial cells
- mass spectrometry
- cell death
- energy transfer
- perovskite solar cells
- pi k akt
- cell proliferation
- liquid chromatography
- structural basis