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Imaging Biological Samples with Optical Microscopy

Imaging Biological Samples with Optical Microscopy

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.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...

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A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

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HiLo microscopy with caustic illumination.

Guorong Hu¹, Joseph Greene¹, Jiabei Zhu¹

¹Department of Electrical & Computer Engineering, Boston University , Boston, Massachusetts 02215, USA.

Biomedical Optics Express

|July 18, 2024

Summary

This summary is machine-generated.

This study introduces caustic HiLo microscopy, a new optical sectioning technique using random caustic patterns. Caustic HiLo microscopy enhances imaging of scattering biological tissues, outperforming traditional speckle-based methods.

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Area of Science:

Biomedical Imaging
Microscopy Techniques
Optical Engineering

Background:

HiLo microscopy combines uniform and structured illumination for optical sectioning.
Random speckle patterns are commonly used for structured illumination in HiLo microscopy.
Speckle patterns offer simplicity and resilience to scattering in biological tissues.

Purpose of the Study:

To introduce and evaluate a novel HiLo microscopy strategy using random caustic patterns.
To compare the performance of caustic HiLo microscopy against speckle HiLo microscopy.
To demonstrate the potential of caustic HiLo for imaging scattering biological samples.

Main Methods:

Developed a HiLo microscopy system utilizing random caustic patterns generated by an off-the-shelf diffuser and a low-coherence LED source.
Employed computational combination of uniform and structured illumination images.
Imaged scattering fixed brain sections of varying thicknesses (100 µm, 300 µm, 500 µm).

Main Results:

Achieved an optical sectioning capability of 4.5 µm with a 20× 0.75 NA objective.
Caustic HiLo microscopy demonstrated superior performance compared to speckle HiLo microscopy.
Showcased enhanced optical sectioning and preservation of fine features in scattering brain tissue.

Conclusions:

Random caustic patterns offer a novel and effective structured illumination strategy for HiLo microscopy.
Caustic HiLo microscopy provides improved optical sectioning and feature preservation in scattering biological samples.
This technique holds promise for diverse biomedical imaging applications.