Measuring Protein Expression in the Rodent Brain Using Near-Infrared Fluorescence and High-Resolution Scanning

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.