Home › Cell Biology › Optical Monitoring of Living Nerve Terminal Labeling in Hair Follicle Lanceolate Endings of the Ex Vivo Mouse Ear Skin
Cell Biology JoVE (Open Access) Citable · DOI

Optical Monitoring of Living Nerve Terminal Labeling in Hair Follicle Lanceolate Endings of the Ex Vivo Mouse Ear Skin

DOI: 10.3791/53855-v
What you'll learn
  • ✓ Prepare ex vivo mouse ear skin for live sensory nerve imaging
  • ✓ Label lanceolate nerve terminals using styryl pyridinium dyes
  • ✓ Image and monitor live nerve terminal staining dynamics
  • ✓ Identify and visualize hair follicle palisade ending morphology
Protocol

Here we describe a novel preparation for imaging live lanceolate sensory terminals of palisade endings that innervate mouse ear skin hair follicles during staining and destaining with styryl pyridinium dyes.

Difficulty
advanced
Total time
~1.5–2 hours per ear preparation and imaging session
Model organism
Mouse (ear skin)
Biosafety
BSL-1

Steps

1
Prepare mouse ear tissue for live labeling

Dissect and isolate mouse ear skin, removing excess tissue to create an ex vivo preparation suitable for optical imaging and dye application to hair follicle nerve terminals.

▶ 01:03
2
Label lanceolate nerve terminals with styryl dyes

Apply styryl pyridinium dyes to the prepared ear tissue to stain living lanceolate sensory terminals of palisade endings innervating hair follicles, monitoring dye uptake in real time.

▶ 03:13
3
Image labeled nerve terminals under optical microscopy

Acquire live fluorescence images of stained lanceolate endings and monitor destaining kinetics as dye is washed out, capturing morphological and kinetic data of sensory terminals.

▶ 06:30
4
Analyze and document labeled nerve terminal morphology

Review and document imaging results showing labeled hair follicle lanceolate endings, confirming successful visualization of palisade terminal structure and dye labeling patterns.

▶ 08:34
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