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

Tunable electrostatic interactions of lipid-coated quantum dots with biological membranes.

bioRxiv : the preprint server for biology·2026
Same author

Dissecting mechanisms of ligand binding and conformational changes in the glutamine-binding protein.

eLife·2026
Same author

AlliGator: Open source fluorescence lifetime imaging analysis in G.

SoftwareX·2026
Same author

High-Performance SiPM Detection Module for Ultra-Fast Time-Resolved Measurements.

Sensors (Basel, Switzerland)·2026
Same author

BLeaching In-cell Single-molecule burstS (BLISS) reveals a small dynamic fraction of HP1α clusters in undifferentiated embryonic stem cells.

bioRxiv : the preprint server for biology·2026
Same author

Real-time wide-field fluorescence lifetime imaging via single-snapshot acquisition for biomedical applications.

PhotoniX·2025

Related Experiment Video

Updated: Feb 28, 2026

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals
07:34

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals

Published on: August 22, 2019

8.5K

16-Ch Time-resolved Single-Molecule Spectroscopy Using Line Excitation.

Antonino Ingargiola1, Pietro Peronio2, Eitan Lerner1

  • 1Dept. Chemistry & Biochemistry, University of California Los Angeles, Los Angeles, CA, USA.

Proceedings of Spie--The International Society for Optical Engineering
|June 13, 2017
PubMed
Summary

This study introduces a novel multispot system for single-molecule spectroscopy, significantly reducing acquisition times. This advancement enables faster detection of biomolecular conformational changes using fluorescence lifetimes.

Keywords:
SPAD arrayTCSPCfluorescencehigh-throughputsingle-molecule

More Related Videos

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

3.0K
Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

10.3K

Related Experiment Videos

Last Updated: Feb 28, 2026

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals
07:34

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals

Published on: August 22, 2019

8.5K
Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

3.0K
Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

10.3K

Area of Science:

  • Biophysics
  • Analytical Chemistry
  • Spectroscopy

Background:

  • Single-molecule spectroscopy on freely-diffusing molecules offers non-perturbing analysis of biomolecular conformational changes.
  • Fluorescence lifetime measurements enhance sensitivity and mitigate artifacts compared to intensity-based methods.
  • Conventional freely-diffusing techniques suffer from prolonged acquisition times.

Purpose of the Study:

  • To develop a time-resolved multispot system to overcome throughput limitations in single-molecule spectroscopy.
  • To enable faster and more efficient detection of biomolecular conformational dynamics.

Main Methods:

  • Implementation of a 16-channel silicon photomultiplier tube (SPAD) array coupled with time-correlated single-photon counting (TCSPC) electronics.
  • Utilization of a shaped 532 nm pulsed laser to create a line excitation pattern, matching the linear detector geometry.
  • Development of a robust and cost-effective multispot system design.

Main Results:

  • The developed system successfully overcomes the long acquisition time issue inherent in freely-diffusing single-molecule spectroscopy.
  • Demonstration of a time-resolved multispot approach with a linear SPAD array.
  • Validation of line-excitation as an effective strategy for multispot implementation.

Conclusions:

  • The novel multispot system significantly enhances throughput for single-molecule spectroscopy.
  • Line-excitation provides a robust and economical method for building multispot systems with linear detector arrays.
  • This technology facilitates more rapid and sensitive studies of biomolecular conformational changes.