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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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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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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Jun 12, 2025

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection
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Subcellular Level Spatial Transcriptomics with PHOTON.

Shreya Rajachandran1,2, Qianlan Xu1,2, Qiqi Cao1,2

  • 1Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX, USA.

Biorxiv : the Preprint Server for Biology
|September 24, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed PHOTON (Photoselection of Transcriptome over Nanoscale), a new method for mapping RNA distribution within cells. This spatial transcriptomic tool reveals RNA

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genomics

Background:

  • RNA localization within subcellular compartments is critical for its function, including storage, processing, translation, and degradation.
  • Understanding the spatial distribution of RNA at a fine resolution is essential for deciphering RNA functions and regulatory mechanisms.

Purpose of the Study:

  • To introduce PHOTON (Photoselection of Transcriptome over Nanoscale), a novel method for spatial transcriptomic profiling at subcellular resolution.
  • To demonstrate the capability of PHOTON in capturing *in situ* transcriptomes of specific cell types and RNA within subcellular compartments.

Main Methods:

  • PHOTON combines high-resolution imaging with high-throughput sequencing.
  • This method enables spatial transcriptome profiling at subcellular resolution.

Main Results:

  • PHOTON accurately captures the transcriptome of target cell types (e.g., ovarian granulosa cells) *in situ*.
  • The method profiles RNA content within subcellular compartments, including the nucleolus and stress granules.
  • PHOTON elucidated the role of m6A modification in mRNA partitioning into stress granules.

Conclusions:

  • PHOTON is a versatile and generalizable platform for spatial transcriptomic analysis.
  • The method provides insights into subcellular molecular dynamics and RNA regulation.