Neuroscience JoVE (Open Access) Citable · DOI

Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection

Name: Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection
Uploaded: 2026-07-13
Duration: 11 min 46 s
Description: Preparation of high-quality RNA from cells of interest is critical to precise and meaningful analysis of transcriptional differences among cell types or ...

DOI: 10.3791/51168-v

What you'll learn

✓ Prepare and stain mouse spinal cord sections for motor neuron isolation
✓ Perform laser capture microdissection to isolate motor neuron cell bodies
✓ Extract high-quality RNA suitable for RNA-seq and qRT-PCR analysis

Protocol

Biopharma Insights High-quality total RNA has been prepared from cell bodies of mouse spinal cord motor neurons by laser capture microdissection after staining spinal cord sections with Azure B in 70% ethanol. Sufficient RNA (~40-60 ng) is recovered from 3,000-4,000 motor neurons to allow downstream RNA analysis by RNA-seq and qRT-PCR.

Difficulty

advanced

Total time

~4–6 hours per spinal cord (including tissue preparation, staining, and microdissection)

Model organism

Mouse (spinal cord)

Biosafety

BSL-1

Steps

Prepare mouse spinal cord sections for analysis

Generate fresh spinal cord tissue sections from mouse tissue. This foundational step ensures optimal morphology and RNA integrity for subsequent staining and microdissection.

▶ 02:23

Stain spinal cord sections with Azure B dye

Apply Azure B stain in 70% ethanol to spinal cord sections to visualize motor neuron cell bodies. This staining enables clear identification of target neurons during laser capture.

▶ 04:21

Isolate motor neurons by laser capture microdissection

Use laser capture microdissection to precisely excise motor neuron cell bodies from stained sections. Recovery of 3,000–4,000 neurons yields 40–60 ng total RNA for downstream analysis.

▶ 07:20

Verify motor neuron isolation and RNA quality

Assess isolated motor neuron samples for purity and RNA integrity. Recovered RNA is suitable for RNA-seq and qRT-PCR applications.

▶ 10:08

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