Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

7.0K
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...
7.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Therapeutic TG2 inhibition reverses systemic multiomic dysregulation in celiac disease.

BMC medicine·2026
Same author

Evaluating MINFLUX experimental performance in silico.

Nature communications·2025
Same author

Multi-step implementation of meiotic crossover patterning.

bioRxiv : the preprint server for biology·2025
Same author

A Photoswitchable HaloTag for Spatiotemporal Control of Fluorescence in Living Cells.

Angewandte Chemie (International ed. in English)·2025
Same author

In-depth single molecule localization microscopy using adaptive optics and single objective light-sheet microscopy.

Nature communications·2025
Same author

RNA-binding proteins mediate the maturation of chromatin topology during differentiation.

Nature cell biology·2025

Related Experiment Video

Updated: Jun 27, 2025

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

8.9K

Build and operation of a custom 3D, multicolor, single-molecule localization microscope.

Rory M Power1, Aline Tschanz2, Timo Zimmermann3,2

  • 1EMBL Imaging Centre, EMBL Heidelberg, Heidelberg, Germany. rory.power@embl.de.

Nature Protocols
|May 3, 2024
PubMed
Summary

Researchers can build their own high-end single-molecule localization microscopy (SMLM) systems. This guide details custom SMLM setup, reducing costs and increasing accessibility for advanced biological imaging.

More Related Videos

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

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

Published on: June 30, 2018

8.5K
Author Spotlight: Advancing Knowledge in Far-From-Equilibrium Materials Through Light-Sheet Microscopy
08:32

Author Spotlight: Advancing Knowledge in Far-From-Equilibrium Materials Through Light-Sheet Microscopy

Published on: January 26, 2024

2.0K

Related Experiment Videos

Last Updated: Jun 27, 2025

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

8.9K
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

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

Published on: June 30, 2018

8.5K
Author Spotlight: Advancing Knowledge in Far-From-Equilibrium Materials Through Light-Sheet Microscopy
08:32

Author Spotlight: Advancing Knowledge in Far-From-Equilibrium Materials Through Light-Sheet Microscopy

Published on: January 26, 2024

2.0K

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Nanotechnology

Background:

  • Single-molecule localization microscopy (SMLM) offers high-resolution biological imaging.
  • Commercial SMLM systems are expensive and complex to develop, limiting accessibility.
  • There is a need for cost-effective, customizable SMLM solutions.

Purpose of the Study:

  • To provide a comprehensive guide for building a custom, high-end SMLM setup.
  • To detail the assembly, alignment, software integration, and operation of a custom SMLM.
  • To enable researchers to establish advanced SMLM capabilities in their labs.

Main Methods:

  • Detailed instructions for optical pathway alignment and hardware/software integration.
  • Validation procedures using test and biological samples with known structures.
  • Guidance on troubleshooting and performance benchmarking of the custom SMLM.

Main Results:

  • A functional, high-end SMLM system can be constructed for $95,000-$180,000.
  • Construction and optimization require an estimated 4-8 months for experienced builders.
  • The guide facilitates the acquisition of high-quality 3D, multicolor super-resolved images.

Conclusions:

  • Building a custom SMLM is feasible and significantly more affordable than commercial options.
  • This guide empowers researchers to implement advanced SMLM for biological discovery.
  • Democratizing access to high-resolution microscopy through open-source, custom-built systems.