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Related Experiment Video

Updated: May 22, 2026

Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy

Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy

Published on: May 1, 2012

Visualization of cortex organization and dynamics in microorganisms, using total internal reflection fluorescence

Felix Spira¹, Julia Dominguez-Escobar, Nikola Müller

¹AG Cellular Dynamics and Cell Patterning, Max Planck Institute of Biochemistry.

Journal of Visualized Experiments : Jove

|May 17, 2012

Summary

Total internal reflection fluorescence (TIRF) microscopy enables high-resolution imaging of cellular dynamics. Bacterial and fungal cell walls enhance TIRF usability, allowing in vivo biochemistry experiments in microorganisms.

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Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Total Internal Reflection Fluorescence (TIRF) microscopy utilizes an evanescent wave for high-resolution imaging near surfaces.
TIRF offers superior axial resolution, high signal-to-noise ratios, and reduced photobleaching compared to other techniques.
Its shallow penetration depth was previously thought to limit applications in cells with thick walls.

Purpose of the Study:

To demonstrate the utility of TIRF microscopy for studying spatio-temporal dynamics in microorganisms.
To highlight how bacterial and fungal cell walls enhance TIRF imaging capabilities.
To provide a practical guide for obtaining high-quality TIRF images in yeast and bacteria.

Main Methods:

Application of TIRF microscopy principles to image fluorescent molecules in vitro and in living cells.
Investigating the interaction of the evanescent wave with microbial cell walls.
Developing protocols for imaging Saccharomyces cerevisiae and Bacillus subtilis using TIRF.

Main Results:

Microbial cell walls improve TIRF usability and expand observable structures.
TIRF enables direct in vivo biochemistry experiments, assessing protein interactions and kinetics in living microorganisms.
Established image restoration techniques can further enhance TIRF image quality.

Conclusions:

TIRF microscopy is highly effective for studying cellular processes in microorganisms.
The unique properties of microbial cell walls are advantageous for TIRF imaging.
This technique facilitates advanced in vivo biochemical studies in single-cell organisms.