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The Blood-brain Barrier00:49

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Updated: Sep 11, 2025

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers
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Blood-brain-barrier permeable fluorescent astrocyte probes.

Nipuni Gunawardhana1, Danielle A Cervasio2, Shangrila Singh1

  • 1Department of Chemistry, Stony Brook University, Stony Brook, New York 11794, United States.

Biorxiv : the Preprint Server for Biology
|August 12, 2025
PubMed
Summary

Researchers developed new, brighter fluorescent probes to visualize brain astrocytes. These advanced astrocyte markers are non-toxic and can cross the blood-brain barrier, aiding in the study of neural circuits.

Keywords:
astrocytesblood-brain-barrierfluorophoresimaging probesmitochondriarodent modelszebrafish

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Astrocytes are crucial glial cells in the central nervous system, maintaining brain homeostasis.
  • Understanding astrocyte roles in neural circuit function requires specific in vivo imaging tools.
  • Existing methods for astrocyte visualization have limitations in specificity, toxicity, or compatibility with genetic techniques.

Purpose of the Study:

  • To develop novel, enhanced fluorescent probes for selective astrocyte labeling.
  • To create brighter, more photostable dyes for improved astrocyte imaging across multiple wavelengths.
  • To assess the probes' subcellular localization and blood-brain barrier penetration.

Main Methods:

  • Synthesis and characterization of a new class of cationic fluorophores.
  • Testing probe specificity, toxicity, and labeling efficiency in rodent and zebrafish models.
  • Confocal microscopy to determine subcellular localization (mitochondria) and assess blood-brain barrier penetration in vivo.

Main Results:

  • The new fluorophores exhibit enhanced brightness and photostability compared to first-generation probes.
  • Probes selectively label astrocytes at low concentrations and short incubation times.
  • Demonstrated mitochondrial localization and successful blood-brain barrier crossing in zebrafish.

Conclusions:

  • This advanced class of astrocyte markers provides superior imaging capabilities for neuroscience research.
  • These probes facilitate visualization and identification of astrocyte subpopulations in various model systems.
  • The developed markers will advance the study of astrocyte contributions to neural circuit function and brain homeostasis.