ENHANCED CYTOCHEMICAL DETECTION OF VIRAL-PROTEINS AND RNAS USING DOUBLE-SIDED LABELING AND LIGHT-MICROSCOPY

We have developed a double-sided labeling technique for detecting vira l proteins or RNAs in plastic-embedded leaf tissue by immunocytochemis try or in situ hybridization, respectively, and light microscopy. The signal from the target was enhanced by double-sided labeling when comp ared with single-sided labeling because sections were submerged in lab eling solutions with both sides accessible to antibodies or complement ary RNAs. The additional label was visible during microscopic analysis . Background signal was decreased since the tissue was probed and wash ed under conditions where folds and creases in the tissue were minimiz ed. This technique uses the same equipment and chemicals as for single -sided labeling, and thus adjustments for reagent expenditures are not necessary. The procedure should be applicable to animal and plant tis sue.