Immunofluorescence database

Database 1 - Examples of NeuN staining for rat and macaque frontal cortex using Protocols A and B

To view database 1, first download the following free program Zeiss LSM Image Browser (this is a Windows based program for PC users). Next, left-click Database 1, which will call up a file download window. Save the compressed database to a local folder on your computer and decompress using WinZip Self-Extractor, StuffIt Expander or other standard software. The folder will then contain a database file (.mdb) and its associated individual .lsm image files. Access the database using Zeiss LSM Image Browser. Detailed information about the confocal microscope settings used for each scan is included (info).

Protocol A produces NeuN staining with a higher signal to noise ratio and better overall quality than protocol B. Examples of images of NeuN staining taken from rat tissue used in this analysis are below.  Additional examples of macaque tissue processed similarly are also provided. Qualitative assessment of the confocal scans from the macaque brains and viewing the tissue directly under a conventional fluorescent microscope reveal a similar relationship to that which was observed for the rat tissue. 

Protocol A	Protocol B	Notes
		Rat 1 NeuN staining in anterior neocortex. These images were obtained by scanning with the settings which brought the maximal intensity of the most brightly labeled cells for either protocol near, but not to, the maximal intensity of 255 greyscale. The settings were held constant throughout the analysis. Some evidence of non-specific labeling is observed in tissue stained using Protocol B when scanned at these settings.
		Rat 1 NeuN staining in anterior neocortex. These images were obtained by scanning with settings optimized for tissue stained using Protocol B.
		Macaque 1 NeuN staining in the prefrontal cortex. These images were obtained by scanning using the settings optimized for tissue processed with Protocol A.
		Macaque 1 NeuN staining in the prefrontal cortex. These images were obtained by scanning using the settings optimized for tissue processed with Protocol B. Using both sets of scanning parameters, Protocol A produced brighter NeuN staiing relative to the background.