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Related Experiment Video

Updated: Jun 25, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

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Published on: May 3, 2022

Multipoint fluorescence correlation spectroscopy with total internal reflection fluorescence microscope.

Yu Ohsugi¹, Masataka Kinjo

¹Hokkaido University, Laboratory of Molecular Cell Dynamics, Faculty of Advanced Life Science, Sapporo 001-0021, Japan.

Journal of Biomedical Optics

|March 5, 2009

Summary

This summary is machine-generated.

This study introduces a multipoint fluorescence correlation spectroscopy (FCS) system for simultaneous diffusion coefficient measurements across cell membranes. The novel setup enables detailed investigation of membrane heterogeneity and molecular dynamics in live cells.

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Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis

Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis

Published on: November 6, 2019

Area of Science:

Biophysics
Cell Biology
Spectroscopy

Background:

Cell membranes exhibit complex, heterogeneous structures.
Understanding molecular dynamics within membranes is crucial for cell function.
Existing methods may lack the spatial resolution for detailed membrane analysis.

Purpose of the Study:

To develop and demonstrate a simultaneous multipoint fluorescence correlation spectroscopy (FCS) system.
To measure diffusion coefficients at multiple locations on a cell membrane concurrently.
To investigate heterogeneous membrane structures and molecular dynamics in live cells.

Main Methods:

Utilized an objective-type total internal reflection FCS (TIR-FCS) system.
Integrated seven optical fibers to create seven detection areas within the evanescent field.
Employed seven photomultiplier tubes (PMTs) and a multichannel correlator for fluorescence intensity fluctuation analysis.

Main Results:

Successfully achieved simultaneous determination of diffusion coefficients at multiple points on cell membranes.
Demonstrated a time resolution of 3 microseconds for the multipoint FCS system.
Enabled investigation of membrane-binding molecular dynamics near glass surfaces in live cells.

Conclusions:

The developed simultaneous multipoint FCS system is effective for studying membrane heterogeneity.
This technique provides high temporal and spatial resolution for analyzing molecular dynamics.
The system offers a valuable tool for live-cell imaging and biophysical studies.