NT & NOS Double

Labeling with antibodies raised against FITC allows amplification of both signals when double labeling. Cy 2⦁ms∝FITC (Jackson) and Dylight 488⦁gt∝FITC (Rockland) have also been tested. All work well (A488 = Cy2 > D488) providing approximately the same degree of signal amplification as Streptavidin.

Double labeling for mouse∝NOS1 (red) and rabbit∝Neurotensin (NT; green), labeled with Cy3⦁Streptavidin and Alexa 488⦁rb∝FITC (Invitrogen), respectively. Component channel images below.

Note that image was acquired as a 4 X 5 2-channel mosaic. Component tiles were exported for each channel. Images of each channel (below) were assembled separately w/ Photomerge--which put the images together slightly differently. Mis-registration can be clearly seen along ventral-lateral surface.

Individual red (NOS) and green (NT) channels. Both received minimum crop and autotone adjustment only. Merged image above cropped and rotated, curve adjustments and unsharp mask applied.