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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...

Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

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 developed.

Overview of Electron Microscopy

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.

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Related Experiment Video

Updated: May 11, 2026

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope

Published on: April 7, 2014

Measuring image resolution in optical nanoscopy.

Robert P J Nieuwenhuizen¹, Keith A Lidke, Mark Bates

¹Quantitative Imaging Group, Delft University of Technology, Delft, The Netherlands.

|April 30, 2013

Summary

This summary is machine-generated.

A new Fourier ring correlation (FRC) method provides a practical resolution measure for optical nanoscopy (super-resolution microscopy). This approach optimizes imaging speed and emitter strategies for better results.

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Last Updated: May 11, 2026

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Area of Science:

Optical Nanoscopy
Super-Resolution Microscopy
Biomedical Imaging

Background:

Current resolution measures in optical nanoscopy are limited.
Existing methods do not account for localization precision, emitter density, and sample structure simultaneously.
A practical, integral resolution metric is needed.

Purpose of the Study:

Introduce a novel resolution measure for optical nanoscopy based on Fourier ring correlation (FRC).
Demonstrate the validity and utility of the FRC method for 2D and 3D localization microscopy images.
Provide a practical tool for optimizing nanoscopy acquisition and analysis.

Main Methods:

Developed a resolution metric using Fourier ring correlation (FRC) directly from image data.
Applied and validated the FRC method on 2D and 3D localization microscopy images of biological structures (tubulin and actin filaments).

Main Results:

The FRC method provides a practical and integral measure of resolution in optical nanoscopy.
Demonstrated its effectiveness on tubulin and actin filament images.
The method allows comparison between different nanoscopy techniques and optimization of labeling strategies.

Conclusions:

The FRC resolution measure offers significant advantages over existing methods.
It enables optimization of data acquisition, emitter localization, and labeling strategies.
Findings suggest prioritizing image resolution speed over solely localization precision.