FIGURE 6.3. Applying RH1691 to the cortex stains the supragranular layers.

FIGURE 6.3

Applying RH1691 to the cortex stains the supragranular layers. (A) Topical application of RH1691 to the mouse barrel cortex labels mainly the supragranular layers. Parasagittal slices (100 μm thick) from stained barrel cortex, counterstained with DAPI and observed with confocal microscopy, reveal that the dye appears to wash out of layer 1, resulting in the highest dye concentrations in layer 2/3. (B) The dye does not enter inside the cells but rather labels membranes. (C) Quantification across different experiments reveals consistent labeling of supragranular layers, with a peak of RH1691 fluorescence in layer 2/3, independent of whether the brain is fixed with paraformaldehyde (PFA) or rapidly frozen and sectioned using a cryostat. (All panels modified from Ferezou, I., Bolea, S., and Petersen, C.C.H. 2006. Neuron 50: 617–629. With permission from Neuron, Cell Press, Elsevier.)

From: Chapter 6, Imaging the Brain in Action: Real-Time Voltage- Sensitive Dye Imaging of Sensorimotor Cortex of Awake Behaving Mice

Cover of In Vivo Optical Imaging of Brain Function
In Vivo Optical Imaging of Brain Function. 2nd edition.
Frostig RD, editor.
Boca Raton (FL): CRC Press/Taylor & Francis; 2009.
Copyright © 2009, Taylor & Francis Group, LLC.

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