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Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Related Experiment Video

Updated: Dec 2, 2025

High-Speed Ultraviolet Photoacoustic Microscopy for Histological Imaging with Virtual-Staining assisted by Deep Learning
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High-Speed Ultraviolet Photoacoustic Microscopy for Histological Imaging with Virtual-Staining assisted by Deep Learning

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Reflection-mode virtual histology using photoacoustic remote sensing microscopy.

Kevan Bell1,2, Saad Abbasi1, Deepak Dinakaran2,3

  • 1Department of Systems Design Engineering, PhotoMedicine Labs, University of Waterloo, E7-6416 200 University Avenue West, Waterloo, ON, N2L 3G1, Canada.

Scientific Reports
|November 6, 2020
PubMed
Summary

Photoacoustic remote sensing (PARS) offers label-free, non-contact imaging of subcellular structures in various tissues. This novel technique accelerates pathology assessment, reducing delays in clinical disease management.

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Area of Science:

  • Biomedical Imaging
  • Histopathology
  • Optical Physics

Background:

  • Current histological methods require extensive tissue preparation, causing significant clinical feedback delays (2-3 weeks).
  • Bright-field microscopy is standard but labor-intensive and time-consuming.
  • Need for rapid, non-invasive imaging techniques in pathology.

Purpose of the Study:

  • To comprehensively study the application of photoacoustic remote sensing (PARS) for label-free tissue visualization.
  • To assess PARS's capability for rapid assessment of large tissue samples and high-contrast imaging.
  • To demonstrate PARS's utility in various histopathological and fresh tissue preparations.

Main Methods:

  • Utilized photoacoustic remote sensing (PARS), a reflection-mode imaging modality.
  • Employed non-contact visualization of intrinsic endogenous optical absorption contrast.
  • Applied PARS to formalin-fixed paraffin-embedded tissues, unstained slides, frozen sections, and fresh tissues.

Main Results:

  • PARS visualized salient subcellular structures in diverse human and murine tissues.
  • The technique provided rapid assessment (<10 min for >1 cm² samples).
  • PARS simulated conventional stains (H&E) and identified pathologies like basal cell carcinoma.

Conclusions:

  • PARS is a versatile, label-free imaging modality for histopathology.
  • It overcomes limitations of traditional tissue preparation, enabling real-time assessment.
  • PARS represents a significant advancement towards a real-time clinical microscope.