Combining Lipophilic dye, in situ Hybridization, Immunohistochemistry, and Histology

Introduce lipophilic dye into target mouse tissue regions for retrograde or anterograde neuronal tracing. This establishes baseline neuronal labeling for subsequent multi-modal analysis.

Apply in situ hybridization probes to detect specific mRNA targets within fixed mouse tissue. This step identifies gene expression patterns complementary to anatomical tracing.

Use antibodies and detection systems to visualize protein localization in the same tissue sections. This adds immunological data to morphological and molecular information.

Mount and process stained sections for light microscopy observation. Standard histological protocols prepare samples for final imaging and data collection.

Combine lipophilic dye, in situ hybridization, immunohistochemistry, and histological findings to characterize neuron populations in mutant mice. Correlate morphology, gene expression, and protein localization in single specimens.