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High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence
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Coherent artifact suppression in line-field reflection confocal microscopy using a low spatial coherence light

Changgeng Liu, Hui Cao, Michael A Choma

    Optics Letters
    |December 23, 2016
    PubMed
    Summary

    Coherent artifacts in line-field reflection confocal microscopy can be reduced using spatially incoherent light sources. This study demonstrates improved noise suppression in scattering samples, highlighting the importance of coherence engineering.

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    Area of Science:

    • Optical Microscopy
    • Coherent Optics
    • Biophotonics

    Background:

    • Line-field reflection confocal microscopy (LF-RCM) offers parallel imaging but can suffer from coherent artifacts due to highly spatially coherent light sources.
    • These artifacts are particularly problematic when imaging scattering biological or material samples.

    Purpose of the Study:

    • To confirm the presence of coherent artifacts in LF-RCM with spatially coherent light.
    • To demonstrate mitigation of these artifacts using reduced spatial coherence light sources.
    • To investigate the impact of source spatial coherence on axial confocality in LF-RCM.

    Main Methods:

    • Utilized a full-pupil LF-RCM system.
    • Employed a vertical-cavity surface-emitting lasers (VCSEL) array as a synthesized reduced spatial coherence light source.
    • Experimentally evaluated performance with highly scattering samples and compared results with a fully coherent light source.

    Main Results:

    • Confirmed that coherent artifacts are a significant issue in LF-RCM with spatially coherent light.
    • Demonstrated substantial suppression of coherent noise in scattering samples by using the VCSEL array light source.
    • Observed that axial confocality in LF-RCM is compromised for scattering samples, irrespective of source spatial coherence.

    Conclusions:

    • Reduced spatial coherence light sources effectively mitigate coherent artifacts in LF-RCM.
    • Spatial coherence engineering is crucial for optimizing parallelized reflection confocal microscopy.
    • Further research may be needed to address the inherent axial confocality limitations with scattering samples.