scholarly journals Fluorescence Lifetime Imaging of pH along the Secretory Pathway

2022 ◽  
Author(s):  
Peter T. A. Linders ◽  
Melina Ioannidis ◽  
Martin ter Beest ◽  
Geert van den Bogaart
Keyword(s):  
Fluorescence Lifetime ◽  
Secretory Pathway ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals Fluorescence lifetime imaging of pH along the secretory pathway

2021 ◽  
Author(s):  
Peter Linders ◽  
Martin ter Beest ◽  
Geert van den Bogaart
Keyword(s):  
Temporal Resolution ◽  
Fluorescence Lifetime ◽  
Secretory Pathway ◽  
Fluorescent Protein ◽  
Excitation Wavelength ◽  
Fluorescence Lifetime Imaging ◽  
Ph Homeostasis ◽  
Ph Changes ◽  
Lifetime Imaging ◽  
Wide Range

Many cellular processes are dependent on correct pH levels, and this is especially important for the secretory pathway. Defects in pH homeostasis in distinct organelles cause a wide range of diseases, including disorders of glycosylation and lysosomal storage diseases. Ratiometric imaging of the pH-sensitive mutant of green fluorescent protein (GFP), pHLuorin, has allowed for targeted pH measurements in various organelles, but the required sequential image acquisition is intrinsically slow and therefore the temporal resolution unsuitable to follow the rapid transit of cargo between organelles. We therefore applied fluorescence lifetime imaging microscopy (FLIM) to measure intraorganellar pH with just a single excitation wavelength. We first validated this method by confirming the pH in multiple compartments along the secretory pathway. Then, we analyze the dynamic pH changes within cells treated with Brefeldin A, a COPI coat inhibitor. Finally, we followed the pH changes of newly-synthesized molecules of the inflammatory cytokine tumor necrosis factor (TNF)-α while it was in transit from the endoplasmic reticulum via the Golgi to the plasma membrane. The toolbox we present here can be applied to measure intracellular pH with high spatial and temporal resolution, and can be used to assess organellar pH in disease models.

Visualization of Cutaneous Distribution of Minocycline of a Topical Gel in Human Facial Skin with Two-Photon Excited Fluorescence Lifetime Imaging Microscopy (FLIM) and Phasor Analysis

2017 ◽  
Vol 1 ◽  
pp. s57
Author(s):  
Sinyoung Jeong ◽  
Maiko Hermsmeier ◽  
Sam Osseiran ◽  
Akira Yamamoto ◽  
Usha Nagavarapu ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Facial Skin ◽  
Topical Gel ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Phasor Analysis

Abstract Not AvailableStudy supported by BioPharmX.

Effects of Photon Losses on Fluorescence Lifetime Imaging Microscopy (FLIM) System Optimization

2015 ◽  
Vol 4 (2) ◽  
pp. 67-77
Author(s):  
Lior Turgeman ◽  
Tsviya Nayhoz ◽  
Nir Roth ◽  
Gilad Yahav ◽  
Avraham Hirshberg ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
System Optimization ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

Phasor S-FLIM: a new paradigm for fast and robust spectral fluorescence lifetime imaging

2021 ◽  
Author(s):  
Lorenzo Scipioni ◽  
Alessandro Rossetta ◽  
Giulia Tedeschi ◽  
Enrico Gratton
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging ◽  
New Paradigm ◽  
Lifetime Imaging

Interrogation of the liver during regeneration by fluorescence lifetime imaging and mass spectrometry

2021 ◽  
pp. 1-1
Author(s):  
Svetlana Rodimova ◽  
Daria Kuznetsova ◽  
Nikolai Bobrov ◽  
Alexander Gulin ◽  
Dmitry Reunov ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

P‐103: Non‐Destructive FMM Shadow‐Level Evaluation with Fluorescence Lifetime Imaging Microscopy

2021 ◽  
Vol 52 (1) ◽  
pp. 1466-1469
Author(s):  
Ki Young Yeon ◽  
Woo Young Lee ◽  
Junghan Shin ◽  
Young Gil Park ◽  
Nari Ahn
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Non Destructive

Fluorescence-Lifetime Imaging of DNA–Dye Interactions within Continuous-Flow Microfluidic Systems

2007 ◽  
Vol 119 (13) ◽  
pp. 2278-2281 ◽  
Author(s):  
Richard K. P. Benninger ◽  
Oliver Hofmann ◽  
Björn Önfelt ◽  
Ian Munro ◽  
Chris Dunsby ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Continuous Flow ◽  
Fluorescence Lifetime Imaging ◽  
Microfluidic Systems ◽  
Lifetime Imaging

scholarly journals Investigation of the Membrane Fluidity Regulation of Fatty Acid Intracellular Distribution by Fluorescence Lifetime Imaging of Novel Polarity Sensitive Fluorescent Derivatives

2021 ◽  
Vol 22 (6) ◽  
pp. 3106
Author(s):  
Giada Bianchetti ◽  
Salome Azoulay-Ginsburg ◽  
Nimrod Yosef Keshet-Levy ◽  
Aviv Malka ◽  
Sofia Zilber ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Membrane Fluidity ◽  
Fluorescence Lifetime ◽  
Intracellular Distribution ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Fatty Acid Derivatives ◽  
Polarity Sensitive

Free fatty acids are essential structural components of the cell, and their intracellular distribution and effects on membrane organelles have crucial roles in regulating the metabolism, development, and cell cycle of most cell types. Here we engineered novel fluorescent, polarity-sensitive fatty acid derivatives, with the fatty acid aliphatic chain of increasing length (from 12 to 18 carbons). As in the laurdan probe, the lipophilic acyl tail is connected to the environmentally sensitive dimethylaminonaphthalene moiety. The fluorescence lifetime imaging analysis allowed us to monitor the intracellular distribution of the free fatty acids within the cell, and to simultaneously examine how the fluidity and the microviscosity of the membrane environment influence their localization. Each of these probes can thus be used to investigate the membrane fluidity regulation of the correspondent fatty acid intracellular distribution. We observed that, in PC-12 cells, fluorescent sensitive fatty acid derivatives with increased chain length compartmentalize more preferentially in the fluid regions, characterized by a low microviscosity. Moreover, fatty acid derivatives with the longest chain compartmentalize in lipid droplets and lysosomes with characteristic lifetimes, thus making these probes a promising tool for monitoring lipophagy and related events.

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