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Updated: Jun 14, 2025
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Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy Conpokal on Live Cells
Published on: August 11, 2020
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Confocal Laser Scanning Platform Combined With In Situ High-Resolution Quantitative Phase Imaging
Wenjing Feng1,2,3, Hongfei Suo1,2,3, Ying Ma1,2,3
1School of Physics, Xidian University, Xi'an, China.
Journal of Biophotonics
|March 7, 2025
Summary
This study reveals that confocal fluorescence microscopy causes less cell damage than wide-field methods. Integrating quantitative phase contrast microscopy with confocal imaging offers a powerful tool for cell biology research.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- Confocal laser scanning microscopy and quantitative phase contrast microscopy are valuable imaging techniques.
- Assessing cellular damage induced by different fluorescence excitation methods is crucial for live-cell imaging.
- Integrating multiple imaging modalities can provide richer biological information.
Purpose of the Study:
- To develop and validate an advanced dual-modality imaging platform combining confocal laser scanning/imaging with flat-fielding quantitative phase contrast microscopy (FF-QPCM).
- To compare the cellular damage caused by confocal fluorescence excitation versus wide-field fluorescence excitation (WFE).
- To investigate the effects of localized confocal laser irradiation on live cells.
Main Methods:
- Integration of a confocal laser scanning/imaging module with FF-QPCM.
- Dual-modality imaging of samples for simultaneous phase and fluorescence data acquisition.
- Comparative analysis of cellular damage (e.g., mitochondrial fragmentation, phase value alterations) after confocal and WFE.
- In situ quantitative phase imaging of live COS7 cells subjected to localized confocal laser irradiation.
Main Results:
- The dual-modality platform achieved high-resolution, high-contrast phase/fluorescence imaging.
- Confocal fluorescence excitation induced minimal cellular damage compared to WFE, which caused significant mitochondrial fragmentation and phase alterations.
- Localized 5-min confocal laser irradiation primarily affected fluorescently labeled mitochondria in COS7 cells, leading to cellular dysfunction.
Conclusions:
- The integrated FF-QPCM and confocal laser scanning/imaging platform is an efficient, sensitive, and information-rich tool for cell biology.
- Confocal excitation is a gentler method for live-cell fluorescence imaging compared to WFE.
- This advanced imaging approach has broad potential for studying cellular dynamics and responses to laser irradiation.
