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Author Spotlight: Efficient Detection of Immune Cell-Infiltration in Cancer Tissues Using Fluorescent Immunohistochemistry

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Cell Cycle Mapping Using Multiplexed Immunofluorescence.

Katarzyna M Kedziora¹, Wayne Stallaert²

¹Department of Cell Biology, Center for Biologic Imaging (CBI), University of Pittsburgh, Pittsburgh, PA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|February 23, 2024

Summary

Cell cycle mapping reveals diverse cell behaviors using advanced imaging and computational analysis. This technique visualizes cell cycle progression and arrest states, offering new insights into cellular heterogeneity.

Keywords:

Cell cycle Immunofluorescence Machine learning Multiplexed imaging Proteomics Single-cell

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Area of Science:

Cell Biology
Computational Biology
Genomics

Background:

Single-cell technologies provide insights into cell cycle regulation.
Heterogeneity in cell cycle progression exists even in identical cell populations.
Understanding cell cycle dynamics is crucial for developmental biology and disease research.

Purpose of the Study:

To describe a general protocol for cell cycle mapping.
To provide a guide for experimental design and analysis in cell cycle studies.
To visualize cell cycle heterogeneity and cell cycle arrest states.

Main Methods:

Cell cycle mapping combines highly multiplexed imaging with manifold learning.
Experimental components include antibody library preparation.
Computational components focus on analysis strategies for visualizing cell paths.

Main Results:

Novel insights into cell cycle commitment and progression.
Evidence of heterogeneity in cell cycle regulation at the single-cell level.
Visualization of diverse cell paths through proliferation and arrest.

Conclusions:

Cell cycle mapping is a powerful tool for studying cellular heterogeneity.
The protocol facilitates the design and analysis of cell cycle experiments.
Understanding single-cell dynamics advances knowledge in cell biology and related fields.