Home › Neuroscience › Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun
Neuroscience JoVE (Open Access) Citable · DOI

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun

DOI: 10.3791/52148-v
What you'll learn
  • ✓ Prepare organotypic brain slices for biolistic targeting
  • ✓ Coat micro and nano-projectiles with DNA
  • ✓ Perform regioselective biolistic transfection on slices
  • ✓ Fix and visualize fluorescent protein expression in slices
Protocol

Biopharma Insights Recent improvements in organotypic brain slice preparations have permitted their exploitation for biotechnological applications. Organotypic slices maintain local structural characteristics of in vivo biology, including functional synaptic connections. Here we present a regioselective biolistic delivery method to label and genetically manipulate these slices.

Difficulty
intermediate
Total time
~1 day
Model organism
Rodent (e.g., rat or mouse)
Biosafety
BSL-1

Steps

1
Prepare organotypic brain slices

Prepare organotypic brain slices from rodent brain, maintaining local structural characteristics and functional synaptic connections.

▶ 01:21
2
Prepare DNA-coated micro and nano-projectiles

Coat micro and nano-projectiles with DNA encoding fluorescent proteins for biolistic delivery.

▶ 02:20
3
Perform biolistic transfection on organotypic slices

Use a modified gene gun to deliver DNA-coated projectiles into organotypic brain slices in a regioselective manner.

▶ 03:38
4
Fix and visualize brain slices

Fix the brain slices and visualize the fluorescent protein expression to confirm successful transfection.

▶ 04:29
5
Analyze biolistic delivery results

Examine the results of biolistic delivery of fluorescent protein encoded DNA into the organotypic slices.

▶ 05:24
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