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Related Experiment Video

Updated: Oct 2, 2025

Long-term Live-cell Imaging to Assess Cell Fate in Response to Paclitaxel

Long-term Live-cell Imaging to Assess Cell Fate in Response to Paclitaxel

Published on: May 14, 2018

Optimizing Long-Term Live Cell Imaging.

Alex Lac¹, Austin Le Lam¹, Bryan Heit^2,3

¹Department of Microbiology and Immunology, The University of Western Ontario, London, ON, Canada.

Methods in Molecular Biology (Clifton, N.J.)

|February 26, 2022

Summary

Minimize photodamage during live cell microscopy for extended observation. This protocol details methods to reduce photobleaching and phototoxicity, enabling precise imaging of cellular processes like endosome trafficking.

Keywords:

Acquisition parameters Fluorescence microscopy Intracellular trafficking Live cell imaging Phagocytosis Photobleaching Photodamage Phototoxicity Time-lapse z-stack

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Area of Science:

Cell biology
Microscopy techniques
Biophysics

Background:

Live cell microscopy is crucial for studying dynamic biological processes.
Fluorescence imaging allows for precise quantification but is limited by photodamage.
Photodamage includes photobleaching of fluorophores and phototoxicity to cells.

Purpose of the Study:

To outline methods for optimizing long-term live cell imaging.
To minimize photodamage (photobleaching and phototoxicity) during imaging.
To demonstrate a protocol for imaging intracellular trafficking of endosomes.

Main Methods:

Utilizing live cell microscopy with fluorescent markers.
Implementing optimized imaging conditions to reduce light exposure.
Employing two- and three-dimensional imaging techniques.
Developing protocols adaptable to various cell types and biological processes.

Main Results:

Successful long-term live cell imaging was achieved.
Photodamage effects were significantly minimized.
Intracellular trafficking of early and late endosomes post-phagocytosis was visualized.
The protocol demonstrated flexibility for diverse biological investigations.

Conclusions:

Optimized live cell microscopy protocols can overcome photodamage limitations.
Minimizing phototoxicity and photobleaching is key for extended live cell imaging.
This adaptable protocol facilitates detailed study of cellular dynamics, including endosome trafficking.