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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Oct 23, 2025

Spatiotemporally Controlled Nuclear Translocation of Guests in Living Cells Using Caged Molecular Glues as Photoactivatable Tags

Spatiotemporally Controlled Nuclear Translocation of Guests in Living Cells Using Caged Molecular Glues as Photoactivatable Tags

Published on: January 17, 2019

Live cell tagging tracking and isolation for spatial transcriptomics using photoactivatable cell dyes.

Alex S Genshaft^1,2,3,4,5,6, Carly G K Ziegler^1,2,3,4,5,6, Constantine N Tzouanas^1,2,3,4,5,6

¹Institute for Medical Engineering & Science, MIT, Cambridge, MA, USA.

Nature Communications

|August 18, 2021

Summary

This summary is machine-generated.

We developed SPACECAT (Spatially PhotoActivatable Color Encoded Cell Address Tags) to track and isolate cells in their microenvironment. This method enables precise analysis of cellular behavior and tissue interactions.

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Area of Science:

Cell Biology
Microenvironment Dynamics
Tissue Engineering

Background:

Cell phenotype and function are dynamically regulated by their microenvironment.
Understanding cellular spatiotemporal activity is crucial for studying tissue (dys)function.

Purpose of the Study:

To develop a method for annotating, tracking, and isolating cells with high spatial and temporal precision.
To enable minimally perturbative analysis of cellular phenotypes within their native microenvironment.

Main Methods:

Developed SPACECAT (Spatially PhotoActivatable Color Encoded Cell Address Tags) using photocaged fluorescent molecules and patterned near-UV light for cell labeling.
Applied SPACECAT to patient-derived intestinal organoids, ex vivo tissue sections from four healthy organs, and a lung tumor model.
Integrated a computational framework for analyzing spatially-biased transcriptome patterns and enriched phenotypes.

Main Results:

SPACECAT demonstrated single-cell precision and temporal stability across diverse cell and tissue types.
Successfully enriched for stem-like and actively mitotic cells in intestinal organoids.
Identified spatially-biased transcriptome patterns and enriched phenotypes in various tissue contexts.

Conclusions:

SPACECAT is a broadly applicable method for linking cellular spatiotemporal and behavioral phenotypes with downstream assays.
This technique provides novel insights into the relationship between tissue microenvironments and cellular (dys)function.
Enables precise investigation of cell-environment interactions in health and disease.