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Contractile dynamics change before morphological cues during fluorescence [corrected] illumination.

S G Knoll¹, W W Ahmed^2,3, T A Saif¹

¹Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA.

Scientific Reports

|December 23, 2015

Summary

Fluorescence microscopy illumination can alter cell contractility almost instantly, detected by quantitative force measurements before visible cell changes occur. This finding is crucial for experiments using live cells and microscopy.

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Area of Science:

Cellular Biophysics
Live-cell Imaging
Mechanobiology

Background:

Fluorescence microscopy is vital for live-cell experiments but can induce unwanted photo-toxic effects.
Assessing light-induced cellular damage currently relies on subjective morphological observations.
Early detection of light-induced cellular stress is needed to ensure experimental validity.

Purpose of the Study:

To quantitatively assess the impact of light exposure on live cell contractility.
To identify early indicators of photo-induced cellular stress before morphological changes are apparent.
To establish a sensitive method for evaluating light-induced cellular responses.

Main Methods:

Fibroblast cells cultured on fluorescent bead-embedded hydrogels to measure substrate deformation.
Traction force microscopy used to quantify cell-generated contractile forces over time.
Controlled exposure to specific wavelengths and intensities of fluorescence illumination.

Main Results:

Cells exposed to moderate fluorescence illumination (540-585 nm, 12.5 W/m²) showed rapid force relaxation within 2 seconds.
Photo-induced morphological changes were not observed for 15-30 minutes post-illumination.
The extent of force relaxation and morphological changes correlated with light wavelength and intensity.

Conclusions:

Cellular contractility changes serve as an early, quantitative indicator of photo-induced stress.
Quantitative force measurements can detect light-induced cellular responses significantly earlier than morphological assessments.
This technique offers a sensitive method to monitor and mitigate light-induced artifacts in live-cell microscopy.