Despite its advantages, RNA ISH has some limitations:
- Technical Complexity: Requires careful optimization of probe design, hybridization conditions, and detection methods. - Tissue Preparation: The quality of tissue fixation and sectioning can affect the sensitivity and specificity of the assay. - Probe Design: Designing probes that specifically bind to the target RNA without cross-reactivity can be challenging. - Signal Detection: The sensitivity of detection methods may limit the ability to detect low-abundance transcripts.
