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

You might also read

Related Articles

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

Sort by
Same author

Suggestions as to the Method of Using Narcotics in Nervous Diseases.

Iowa medical journal·2023
Same author

Suggestions as to the Method of Using Narcotics in Nervous Diseases.

Iowa medical journal·2023
Same author

Introductory Lecture to the Class of the Iowa University.

Iowa medical journal·2023
Same author

Human Health on the Moon and Beyond and the Results of the Spaceflight for Everybody Symposium.

Aerospace medicine and human performance·2023
Same author

Cheap Dentistry and Its Tendencies.

The Dental register·2021
Same author

Efficient Front-Rear Coupling in Neutrophil Chemotaxis by Dynamic Myosin II Localization.

Developmental cell·2019
Same journal

Lower bound of the capacitance of constant phase elements based on electrochemical impedance spectra.

Physical chemistry chemical physics : PCCP·2026
Same journal

Stability constants of lanthanide-nitrate complexes in aqueous solutions: a theoretical study.

Physical chemistry chemical physics : PCCP·2026
Same journal

Lead-free Cs<sub>3</sub>MnCl<sub>5</sub> and CsMnCl<sub>3</sub> crystals: rapid on-chip crystallization, phase transition and fluorescence sensing applications.

Physical chemistry chemical physics : PCCP·2026
Same journal

F-Interstitial passivation preserves host-like optoelectronic properties in <sup>229</sup>Th:YLF nuclear-clock platforms.

Physical chemistry chemical physics : PCCP·2026
Same journal

Structural trends of tryptophan dimer: hydrogen bonding <i>versus</i> π-stacking from an energy decomposition analysis perspective.

Physical chemistry chemical physics : PCCP·2026
Same journal

Achieving high thermoelectric performance in Sb<sub>2</sub>Se<sub>3</sub>-alloyed GeTe through synergistic optimization of electrical and thermal transport.

Physical chemistry chemical physics : PCCP·2026
See all related articles

Related Experiment Video

Updated: May 7, 2026

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

Sample preparation for single molecule localization microscopy.

John R Allen1, Stephen T Ross, Michael W Davidson

  • 1National High Magnetic Field Laboratory and Department of Biological Science, The Florida State University, 1800 East Paul Dirac Drive, Tallahassee, 32304, Florida, USA. davidson@magnet.fsu.edu.

Physical Chemistry Chemical Physics : PCCP
|October 3, 2013
PubMed
Summary
This summary is machine-generated.

Proper sample preparation is crucial for single-molecule superresolution microscopy. This guide details live/fixed cell prep, fluorophore choice, and buffer considerations for optimal imaging sensitivity.

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

Single-Molecule Localization Microscopy of Membrane Proteins using Single-Antibody Labeling
07:51

Single-Molecule Localization Microscopy of Membrane Proteins using Single-Antibody Labeling

Published on: March 20, 2026

Related Experiment Videos

Last Updated: May 7, 2026

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

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

Single-Molecule Localization Microscopy of Membrane Proteins using Single-Antibody Labeling
07:51

Single-Molecule Localization Microscopy of Membrane Proteins using Single-Antibody Labeling

Published on: March 20, 2026

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • Single molecule localization microscopy (SMLM) offers nanoscopic resolution, surpassing traditional light microscopy limits.
  • SMLM technology has matured, with commercial systems available, yet sample preparation remains a critical, often overlooked, factor.
  • Effective SMLM relies heavily on meticulous sample preparation for achieving single-molecule sensitivity.

Purpose of the Study:

  • To provide a comprehensive analysis of sample preparation techniques for SMLM.
  • To address the gap in resources concerning sample preparation for high-sensitivity superresolution imaging.
  • To guide researchers in optimizing sample preparation for various SMLM applications.

Main Methods:

  • Detailed analysis of sample preparation protocols for both live and fixed cells.
  • Evaluation of critical parameters including fluorophore selection and staining methodologies.
  • Consideration of imaging buffer compositions essential for single-molecule detection.

Main Results:

  • Identification of key factors influencing SMLM image quality and sensitivity.
  • Demonstration of how specific preparation choices impact localization precision and resolution.
  • Highlighting the importance of optimizing each step from cell handling to buffer formulation.

Conclusions:

  • Optimized sample preparation is paramount for successful single-molecule superresolution imaging.
  • Researchers must carefully consider fluorophore choice, fixation/staining, and buffer conditions for optimal results.
  • This work serves as a vital resource for advancing SMLM applications through improved sample preparation.