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Updated: Jul 31, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Quantitative Single-Molecule Localization Microscopy.

Siewert Hugelier¹, P L Colosi¹, Melike Lakadamyali^1,2,3

¹Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; email: melikel@pennmedicine.upenn.edu, Siewert.Hugelier@Pennmedicine.upenn.edu, pcolosi@pennmedicine.upenn.edu.

Annual Review of Biophysics

|May 9, 2023

Summary

This summary is machine-generated.

Super-resolution microscopy offers nanoscale insights into cells. This review details methods for quantifying data from techniques like single-molecule localization microscopy (SMLM) for robust biological analysis.

Keywords:

quantitative analysis quantitative cell biology single-molecule localization microscopy super-resolution nanoscopy

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Last Updated: Jul 31, 2025

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Area of Science:

Cellular and Molecular Biology
Biophysics
Optical Microscopy

Background:

Super-resolution fluorescence microscopy enables nanoscale visualization of cellular structures.
Advancements focus on reliable quantification of biological data obtained from these techniques.

Purpose of the Study:

To review the principles of super-resolution microscopy techniques, specifically STED and SMLM.
To provide an overview of methodological developments for quantifying super-resolution data, with an emphasis on SMLM.

Main Methods:

Description of fundamental super-resolution microscopy principles (STED, SMLM).
Overview of quantification methods: spatial point pattern analysis, colocalization, protein copy number quantification.
Exploration of advanced techniques: structural modeling, single-particle tracking, biosensing.

Main Results:

Comprehensive review of established and emerging quantitative methods for super-resolution microscopy data.
Detailed explanation of techniques applicable to single-molecule localization microscopy (SMLM).

Conclusions:

Quantitative super-resolution microscopy is crucial for robust biological data analysis.
Future research directions in applying quantitative super-resolution microscopy to new biological questions are highlighted.