Visualizing Astrocyte Morphology Using Fluorescent Dye Iontophoresis in a Fixed Mouse Brain Slice

Abstract

Source: Moye, S. L., et.al. Visualizing Astrocyte Morphology Using Lucifer Yellow Iontophoresis. J. Vis. Exp. (2019).

This video demonstrates the use of fluorescent dye iontophoresis to study astrocyte morphology in fixed brain slices. It details the piercing of the astrocyte soma with an electrode to deliver an anionic, hydrophilic dye into the cytoplasm under a low-voltage current, enabling the dye to diffuse and label the soma and branches. Confocal microscopy is then used to visualize the labeled astrocytes, providing morphological insights.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Borosilicate glass pipettes with filament	World precision instruments	1B150F-4
Confocal laser-scanning microscope	Olympus	FV1000MPE	A similar alternative can be used
Imaris software (Version 7.6.5)	Bitplane Inc.		A similar alternative can be used
Lucifer Yellow CH dilithium salt	Sigma	L0259
Lucifer Yellow CH dipotassium salt	Sigma	L0144
Objective lens (40x)	Olympus	LUMPLFLN 40XW	A similar alternative can be used
Objective lens (60x)	Olympus	PlanAPO 60X	A similar alternative can be used
PBS tablets, 100 mL	VWR	VWRVE404	An identical alternative can be used
Pipette micromanipulator- Model ROE-200	Sutter	MP-285 / ROE-200 / MPC-200	A similar alternative can be used
Potassium Chloride	Sigma	P3911	An identical alternative can be used
Stimulator- Model Omnical 2010	World precision instruments	Omnical 2010	A similar alternative can be used