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Imaging Biological Samples with Optical Microscopy01:18

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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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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Phase Contrast and Differential Interference Contrast Microscopy01:26

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Phase-Contrast Microscopes
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The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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Updated: Nov 10, 2025

Integrated Photoacoustic Ophthalmoscopy and Spectral-domain Optical Coherence Tomography
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Microscope integrated optical coherence tomography system combined with augmented reality.

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    This study introduces a guided laser registration method for microscope-integrated optical coherence tomography (MI-OCT) systems. It enables accurate 3D-2D image matching for improved surgical visualization in ophthalmic procedures.

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    Area of Science:

    • Ophthalmic surgery
    • Medical imaging
    • Augmented reality

    Background:

    • Current microscope-integrated optical coherence tomography (MI-OCT) systems display images from different modalities independently.
    • Surgeons face challenges in subjectively matching 2D and 3D images of the same surgical area.
    • This subjective matching can lead to inaccuracies in surgical guidance.

    Purpose of the Study:

    • To develop a simple registration method for MI-OCT systems to overcome independent image display limitations.
    • To integrate augmented reality with MI-OCT using a guided laser for improved image registration.
    • To enhance surgical precision by accurately aligning 2D microscopic and 3D OCT data.

    Main Methods:

    • A guided laser system was introduced into the MI-OCT setup to identify fiducials in microscopic images.
    • Fiducial coordinates in the OCT model were calculated using applied voltages of the scanning galvanometer mirror.
    • Homography matrix and camera parameters were employed to superimpose the reconstructed 3D OCT model onto 2D microscopic images.

    Main Results:

    • The method successfully generated 2D microscopic images with depth information from scanning regions.
    • Experiments were conducted using artificial and animal eyes to validate the technique.
    • A registration error of 0.04 mm was achieved, falling within acceptable surgical limits.

    Conclusions:

    • The proposed guided laser registration method effectively aligns 2D microscopic and 3D OCT images.
    • This technique significantly reduces subjective error in image matching for MI-OCT systems.
    • The method holds substantial potential for various ophthalmic surgical applications, improving visualization and accuracy.