What Are the Challenges in Fluorescence Multiplexing?
Despite its advantages, fluorescence multiplexing faces several challenges:
Spectral Overlap: Ensuring minimal overlap between the emission spectra of different fluorochromes to avoid signal bleed-through. Photobleaching: Fluorochromes can lose their fluorescence over time under light exposure, which can affect signal intensity and data accuracy. Tissue Autofluorescence: Some tissues have intrinsic fluorescence that can interfere with the detection of specific fluorochromes. Complexity: The need for precise optimization of staining protocols and imaging conditions to achieve reliable results.
