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Biofilms

Biofilms

Biofilms are complex communities of microorganisms encased in a self-produced extracellular polysaccharide matrix attached to surfaces. These microbial consortia can include single or multiple species, providing enhanced survival benefits by forming organized, multilayered structures.The formation of biofilms occurs through four key stages: attachment, colonization, development, and dispersal.During attachment, free-swimming planktonic cells adhere to a surface, often facilitated by...

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Related Experiment Video

Updated: Dec 25, 2025

Clarifying and Imaging Candida albicans Biofilms

Clarifying and Imaging Candida albicans Biofilms

Published on: March 6, 2020

Clarifying and Imaging Candida albicans Biofilms.

Frederick Lanni¹, Katherine Lagree², Manning Y Huang²

¹Department of Biological Sciences, Carnegie Mellon University; lanni@cmu.edu.

Journal of Visualized Experiments : Jove

|March 24, 2020

Summary

Candida albicans biofilms are difficult to study optically. This research developed a novel clarification method using methyl salicylate for improved confocal microscopy of fungal biofilms.

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Last Updated: Dec 25, 2025

Clarifying and Imaging Candida albicans Biofilms

Clarifying and Imaging Candida albicans Biofilms

Published on: March 6, 2020

Visualization of Biofilm Formation in Candida albicans Using an Automated Microfluidic Device

Visualization of Biofilm Formation in Candida albicans Using an Automated Microfluidic Device

Published on: December 14, 2017

A Soluble Tetrazolium-Based Reduction Assay to Evaluate the Effect of Antibodies on Candida tropicalis Biofilms

A Soluble Tetrazolium-Based Reduction Assay to Evaluate the Effect of Antibodies on Candida tropicalis Biofilms

Published on: September 16, 2022

Area of Science:

Mycology
Microbiology
Biophysics

Background:

Candida albicans transitions from commensal to virulent states, forming biofilms on surfaces.
Biofilms exhibit antifungal drug resistance and can cause systemic infections.
Optical microscopy is hindered by biofilm opacity and refractive heterogeneity.

Purpose of the Study:

To develop a method for visualizing internal structures of intact Candida albicans biofilms.
To enable detailed study of biofilm formation and cellular morphology.

Main Methods:

Developed a stepwise solvent exchange protocol for refractive index matching.
Utilized methyl salicylate (n=1.537) for optimal biofilm clarification.
Employed confocal fluorescence microscopy to visualize clarified biofilms.

Main Results:

Achieved visualization of 600 µm intact biofilms from apex to base with minimal signal attenuation.
Successfully imaged internal structural, cellular, and subcellular features.
Demonstrated the utility of the protocol for studying fungal biofilms.

Conclusions:

Refractive index matching via solvent exchange is effective for optical clarification of fungal biofilms.
This method significantly enhances the capability of confocal microscopy for biofilm research.
The protocol provides a valuable tool for understanding Candida albicans biofilm dynamics and structure.