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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...

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Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

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Total Internal Reflection Fluorescence Microscopy

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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Related Experiment Video

Updated: Apr 29, 2026

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Every laboratory with a fluorescence microscope should consider counting molecules.

Valerie C Coffman¹, Jian-Qiu Wu²

¹Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210 coffman.147@osu.edu wu.620@osu.edu.

Molecular Biology of the Cell

|May 15, 2014

Summary

This summary is machine-generated.

Accurately counting protein molecules in cells using fluorescence microscopy is crucial for understanding biological processes. This essay promotes awareness and adoption of protein quantification methods in cell biology research.

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Last Updated: Apr 29, 2026

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

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Published on: October 28, 2018

Area of Science:

Cell Biology
Biophysics
Biochemistry

Background:

Protein abundance is a key determinant of cellular function and dynamics.
Accurate protein quantification is essential for various biological applications, from understanding reaction kinetics to building predictive models.
Current methods for protein counting offer varying levels of utility and precision.

Purpose of the Study:

To raise awareness of fluorescence microscopy techniques for quantifying protein molecules within cells.
To encourage the broader adoption and reporting of protein numbers in cell biology studies.
To provide an overview of the current state, accuracy, and utility of protein quantification methods.

Main Methods:

Utilizes fluorescence microscopy as the primary tool for protein molecule counting.
Discusses various established and emerging techniques for single-cell protein quantification.
References detailed protocols and applications for specific methodologies.

Main Results:

Highlights the critical role of precise protein numbers in understanding cellular mechanisms.
Demonstrates the feasibility and importance of quantitative protein measurements.
Provides a framework for evaluating the accuracy and applicability of different quantification approaches.

Conclusions:

Accurate protein quantification via fluorescence microscopy is vital for advancing cell biology.
Increased reporting of protein numbers will enhance reproducibility and deepen biological insights.
The field benefits from a unified approach to protein molecule counting and data reporting.