The Neuro Probe P48TM template is a thin piece of plastic with a pattern of holes identical to the pattern of wells in a Neuro Probe AP48 48-well chemotaxis chamber. The template is used to help locate the spots of cells on the filter after the filter has been stained. For most assays it is not necessary to use the template since there are usually many cells, and the stain is very visible. But if there is very little migration of cells, or if the stain used is not visible, it can be difficult to distinguish cell sites on the filter from spaces between sites.
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AP48 48-well chemotaxis chambers have two small holes in the top plate situated on the mid-line. These holes match corresponding holes in the template and are used to position the template in relation to the filter.
Assemble the chamber with the filter in place. Cut the cotton end off a cotton swab that has a cardboard — not plastic — stem, wet the end with an indelible magic marker, insert the marked end through each of the two small holes in the top plate and rotate it gently against the filter to make a round spot. Do not press too hard, or the filter will be displaced or punctured. If necessary pare down the cotton swab to fit the hole. The filter now has two spots on the midline that will match the midline holes in the template.
After the assay is complete and the filter is stained and flattened with immersion oil on a glass microscope slide (see AP48 protocol) align the spots on the filter with the midline holes in the template. The template frames the cell sites, and the filter is ready to be viewed under a microscope.