Neuroscience JoVE (Open Access) Citable · DOI

Vibrodissociation of Neurons from Rodent Brain Slices to Study Synaptic Transmission and Image Presynaptic Terminals

Name: Vibrodissociation of Neurons from Rodent Brain Slices to Study Synaptic Transmission and Image Presynaptic Terminals
Uploaded: 2026-08-06
Duration: 8 min 38 s
Description: Mechanical dissociation of neurons from the central nervous system has the advantage that presynaptic boutons remain attached to the isolated neuron of ...

DOI: 10.3791/2752-v

What you'll learn

✓ Prepare viable neurons with intact presynaptic boutons using vibrodissociation
✓ Perform patch-clamp electrophysiological recording on isolated neurons
✓ Image presynaptic terminals and measure synaptic transmission properties
✓ Apply pharmacological manipulations with improved experimental control

Protocol

Biopharma Insights This report demonstrates a technique for mechanical isolation of individual viable neurons retaining attached presynaptic boutons. Vibrodissociated neurons have the advantages of rapid production, excellent pharmacological control and improved space-clamp without influence from neighboring cells. This method can be used for imaging of synaptic elements and patch-clamp recording.

Difficulty

advanced

Total time

~3–4 hours per experiment (slice preparation ~30 min, dissociation ~20 min per slice, recording/imaging ~2–3 hours)

Model organism

Rodent (mouse/rat) brain tissue

Biosafety

BSL-1

Steps

Prepare acute brain slices from rodent

Extract and slice rodent brain tissue to obtain viable acute brain slices suitable for enzymatic and mechanical dissociation. Maintain slice viability in oxygenated artificial cerebrospinal fluid.

▶ 02:14

Prepare flame-sealed glass micropipettes

Fabricate flame-sealed micropipettes and perform mechanical vibrodissociation of neurons from brain slices. This step produces individual neurons with attached presynaptic boutons.

▶ 03:22

Perform patch-clamp electrophysiological recording

Establish whole-cell or other patch-clamp configurations on dissociated neurons to measure membrane currents and synaptic transmission properties with pharmacological control.

▶ 05:14

Image presynaptic terminals and analyze synaptic transmission

Visualize presynaptic boutons and record synaptic transmission data from vibrodissociated neurons. Representative results demonstrate imaging quality and electrophysiological readouts.

▶ 06:39

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