Real-Time Label-Free Imaging and Quantitative Analysis of Macrophage Morphodynamics Using Optical Diffraction...

Area of Science:

• Cell Biology
• Biophysics
• Microscopy

Background:

• Live-cell imaging is crucial for understanding dynamic cellular processes.
• Traditional methods often require exogenous labeling, which can affect cell behavior.
• Observing morphological changes and motility in real-time is essential for studying cell responses.

Purpose of the Study:

• To establish a label-free workflow using optical diffraction tomography (ODT) for live-cell imaging and analysis.
• To enable continuous, long-term observation of single-cell morphology and movement under stable conditions.
• To quantify dynamic cellular behaviors, such as projected area, perimeter, and migration speed.

Main Methods:

• Developed a label-free optical diffraction tomography (ODT)-based workflow.
• Performed time-lapse imaging of RAW264.7 macrophages under lipopolysaccharide stimulation and baicalin pretreatment.
• Utilized ODT for quantitative analysis of single-cell morphology and motility over extended periods.

Main Results:

• The ODT workflow enabled stable, long-term tracking of individual macrophages.
• Quantitative parameters like projected area, perimeter, and migration speed were extracted.
• The method successfully reflected temporal changes in cell morphology and motility in response to stimuli.

Conclusions:

• The label-free ODT workflow provides a practical approach for observing dynamic cellular behaviors without exogenous labeling.
• This method complements conventional assays by offering continuous, quantitative insights into cell morphodynamics.
• The workflow is potentially applicable to other adherent cell types for studying cellular responses to various conditions.