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Updated: Aug 15, 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

Single-molecule localization microscopy goes quantitative.

Silvia Scalisi^1,2, Dario Pisignano³, Francesca Cella Zanacchi^2,3

¹Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Trento, Italy.

Microscopy Research and Technique

|January 5, 2023

Summary

This summary is machine-generated.

Quantitative super-resolution microscopy enables precise protein counting at the nanoscale. This primer explores methods like stepwise photobleaching and calibration standards for studying protein complexes and overcoming technical challenges.

Keywords:

fluorescence microscopy quantitative stochastic optical reconstruction microscopy (STORM)single-molecule localization super-resolution microscopy

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Last Updated: Aug 15, 2025

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

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Published on: June 30, 2018

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Published on: August 22, 2017

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Single-molecule localization microscopy (SMLM) is increasingly used in biological research.
Super-resolution microscopy is emerging as a quantitative tool for nanoscale protein quantification.
Understanding protein complexes requires precise measurement of protein copy numbers.

Purpose of the Study:

To introduce the fundamental concepts of quantitative super-resolution microscopy.
To provide strategies for applying quantitative SMLM to biological questions.
To address technical challenges in nanoscale protein quantification.

Main Methods:

Stepwise photobleaching for protein counting.
Quantitative SMLM using calibration standards.
Analysis of protein complexes at the nanoscale.

Main Results:

Demonstration of SMLM as a quantitative method for protein copy number determination.
Successful application of quantitative approaches to study protein complexes.
Identification of strategies to overcome limitations in quantitative super-resolution microscopy.

Conclusions:

Quantitative super-resolution microscopy offers powerful insights into protein complexes.
Stepwise photobleaching and calibration standards are key to accurate nanoscale quantification.
This primer equips researchers with knowledge to apply these advanced techniques effectively.