Confocal laser scanning platform combined with in situ high-resolution
quantitative phase imaging
Abstract
This paper introduces an advanced dual-modality imaging platform by
integrating a confocal laser scanning/imaging module with flat-fielding
quantitative phase contrast microscopy (FF-QPCM). The platform enables
phase/fluorescence dual-modality imaging of the same samples with high
resolution and contrast. Using this platform, we observed that confocal
fluorescence excitation causes minimal damage to cells, whereas
wide-field fluorescence excitation results in significant damage,
particularly mitochondrial fragmentation and notable alterations in
phase values. Additionally, in situ quantitative phase imaging was
conducted on live COS7 cells with a specific region irradiated by a
confocal laser. The results demonstrated that 5-minute continuous
confocal laser irradiation affects only COS7 cells with fluorescently
labeled mitochondria, causing mitochondrial dysfunction throughout the
cell. These findings suggest that integrating FF-QPCM with a confocal
laser scanning/imaging module offers a highly efficient, sensitive, and
information-rich tool for cell biology research, with broad application
potential.