High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals
Figure 1
Proteinase K treatment is necessary to allow QD penetration and labeling of deep tissues in Xenopus and QD705 nm anti-Dig conjugates can be used for the detection of transcripts in wholemount in situ hybridization experiments. (a) Detection of biotinylated 4G10 (anti-Phosphotyrosine) antibody using streptavidin conjugated 655 nm QDs in a triton permeabilized embryo. Specific staining can be seen at the cell-cell boundaries of the ectodermal cells of the fin (upper part of the image) but not in the deep tissues (somites at the bottom part of the image). (b) Detection of biotinylated 4G10 (anti-Phosphotyrosine) antibody using streptavidin conjugated 655 nm QDs in a PK permeabilized embryo. Specific staining of the deep intersomitic boundaries can be seen. Superficial cells of the fin cannot be seen due to degradation of this delicate structure by the PK treatment. (c) QD705 nm anti-DIG antibody labeling of the probe for LTBP1 generates a staining pattern that closely matches the published expression for this mRNA [1]. QDs label the somites as well as anterior neural and neural crest tissues including the branchial arches and a region surrounding the eye. (d) Imaging of the QD labeling for LTBP1 in the somites at 10X magnification.
