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Related Experiment Video

Updated: May 13, 2026

Quantifying Cytoskeleton Dynamics Using Differential Dynamic Microscopy

Quantifying Cytoskeleton Dynamics Using Differential Dynamic Microscopy

Published on: June 15, 2022

Quantitative dynamic footprinting microscopy.

Prithu Sundd¹, Klaus Ley

¹Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037, USA.

Immunology and Cell Biology

|March 13, 2013

Summary

Quantitative dynamic footprinting (qDF) visualizes leukocyte rolling. Dual color qDF (DqDF) advances this technique, enabling 3D footprint analysis for studying cell adhesion under flow conditions.

Area of Science:

Cellular and Molecular Biology
Biophysics
Immunology

Background:

Leukocyte adhesion under flow is critical in inflammatory responses.
Understanding the mechanics of leukocyte rolling is essential for diagnosing and treating inflammatory diseases.
Existing techniques have limitations in visualizing the dynamic interactions of rolling leukocytes.

Purpose of the Study:

To introduce and detail the dual color quantitative dynamic footprinting (DqDF) technique.
To demonstrate the capability of DqDF in visualizing leukocyte footprints with high resolution.
To explore the application of DqDF in elucidating mechanisms of leukocyte adhesion at high shear stress.

Main Methods:

Quantitative dynamic footprinting (qDF) based on total internal reflection fluorescence.

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Dual color qDF (DqDF) for simultaneous visualization of two fluorochromes.

3D reconstruction of leukocyte footprints from 2D qDF images.

Main Results:

DqDF enables simultaneous visualization of two fluorochromes in leukocyte footprints.
The technique allows for the creation of 3D topographical renditions of cell footprints.
DqDF has been successfully applied to study neutrophil rolling mechanisms under inflammatory shear stresses.

Conclusions:

DqDF is a powerful advancement over qDF for studying leukocyte adhesion dynamics.
The technique provides novel insights into the mechanisms governing leukocyte rolling under physiological flow conditions.
DqDF offers a valuable tool for research in immunology and inflammatory disease.