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Confocal Fluorescence Microscopy

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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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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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Updated: May 3, 2026

Confocal and Super-Resolution Imaging of Polarized Intracellular Trafficking and Secretion of Basement Membrane Proteins During Drosophila Oogenesis

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Confocal supercritical angle microscopy for cell membrane imaging.

Siddharth Sivankutty, Thomas Barroca, Céline Mayet

|February 4, 2014

Summary

This summary is machine-generated.

We achieved subwavelength resolution in biological samples using supercritical angle fluorescence with a standard confocal microscope. This technique enhances surface imaging, particularly for cell membranes, by utilizing a simple modification to the microscope

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Confocal and Super-Resolution Imaging of Polarized Intracellular Trafficking and Secretion of Basement Membrane Proteins During Drosophila Oogenesis

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Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

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Published on: February 24, 2026

Area of Science:

Biophysics
Microscopy
Optical Physics

Background:

Confocal microscopy offers optical sectioning capabilities.
High-resolution imaging of biological samples, especially surfaces like cell membranes, remains a challenge.
Supercritical angle fluorescence (SAF) utilizes light emitted at angles exceeding the critical angle for total internal reflection.

Purpose of the Study:

To demonstrate subwavelength sectioning on biological samples using a modified conventional confocal microscope.
To integrate the phenomenon of supercritical angle fluorescence into a practical microscopy technique.
To enable high-resolution surface imaging of biological specimens.

Main Methods:

Utilized a conventional confocal scanning microscope.
Integrated a simple modification to the detection channel to capture supercritical angle fluorescence.
Exploited the emission of propagating radiation into "forbidden angles" by fluorophores at refractive index discontinuities.

Main Results:

Achieved subwavelength resolution in optical sectioning of biological samples.
Demonstrated the feasibility of integrating SAF with standard confocal microscopy.
Showcased the potential for high-resolution surface imaging.

Conclusions:

Confocal-supercritical angular fluorescence microscopy enables subwavelength resolution.
The technique is easily adaptable to existing high numerical aperture confocal microscopes.
This method is a powerful tool for high-resolution surface and membrane imaging in biological samples.