JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:03 min • May 29th, 2025
Take a glass slide containing rat brain tissue sections immunostained for AMPA and NMDA receptors in amygdala neurons.
AMPA receptors mediate glutamate-induced sodium influx and fast excitatory synaptic transmission.
In contrast, NMDA receptors mediate glutamate and co-agonist-induced sodium and calcium influx, facilitating slow excitatory synaptic transmission.
The receptors are stained with primary antibodies and detected using near-infrared fluorophore-tagged secondary antibodies.
Place the slide tissue-side down on the near-infrared scanning interface.
Adjust the imaging parameters and begin imaging.
Upon laser illumination, the fluorophores on the AMPA and NMDA receptors emit fluorescence.
Near-infrared imaging minimizes autofluorescence, enhancing signal clarity.
High-resolution scanning further ensures clear differentiation of fluorescence signals, enabling precise visualization of nearby AMPA and NMDA receptors in the amygdala.
Using imaging software, calculate the AMPA/NMDA fluorescence intensity ratio in a specific region of interest as a marker of learning and memory.
Place slides onto near-infrared scanning interface with the tissue facing down. Image multiple slides at a time
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