Home › Neuroscience › Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo
Neuroscience JoVE (Open Access) Citable · DOI

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

DOI: 10.3791/3921-v
What you'll learn
  • ✓ Perform retrograde fluorescent labeling to identify projection-specific neurons in vivo
  • ✓ Visualize labeled axons and target them for extracellular single-unit recording
  • ✓ Record neural activity from identified neurons without genetic manipulation
Protocol

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

Difficulty
advanced
Total time
~4-6 hours per animal (including surgery, labeling, visualization, and recording)
Model organism
Not specified in abstract; likely rodent based on in vivo surgery/recording context
Biosafety
BSL-1

Steps

1
Prepare surgical site and animal

Prepare the animal and surgical field for intracranial injection and recording procedures. This includes positioning, anesthesia, and sterile preparation.

▶ 02:26
2
Perform surgical access to target region

Conduct surgical procedures to expose the brain region of interest for retrograde labeling injection and subsequent extracellular recording.

▶ 04:08
3
Inject retrograde fluorescent dye and visualize axons

Inject fluorescent tracer at the axonal projection target site to label neurons via retrograde transport, then identify and visualize labeled axons under fluorescence microscopy.

▶ 06:10
4
Record extracellular activity from identified axons

Position extracellular electrodes on visually identified fluorescently-labeled axons and acquire single-unit neural recordings.

▶ 08:27
5
Analyze and interpret representative recordings

Review and present example extracellular recordings obtained from retrograde-labeled neurons, demonstrating successful identification and activity characterization.

▶ 10:58
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