Quantitation of cellular resolution in situ hybridization histochemistry in brain by image analysis.
A new method for relative quantitation of mRNA levels with single neuron resolution is described. Hybridized tissue sections are emulsion dipped, exposed, and developed. The resulting silver grains are visualized using dark-field microscopy at high magnification. The method relies on computer-based image analysis of sequentially located, high resolution, small fields containing the cell images, each of which is analyzed for mRNA content. Maps of cell distributions are constructed, with cell marks color-coded for relative mRNA levels, yielding previously unavailable information on regional distribution of quantitative cellular expression of specific mRNA in brain.
Engineering Department, E.I. Du Pont de Nemours and Co., Inc., Wilmington, DE 19898.
W T
WT
Rogers
J S
JS
Schwaber
M E
ME
Lewis
D001923:Brain Chemistry; D006651:Histocytochemistry; D007091:Image Processing, Computer-Assisted; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger