- Spectral Overlap: One significant challenge is the spectral overlap between different fluorescent signals, which can complicate the interpretation of images. - Photobleaching: The tendency of fluorescent materials to degrade under prolonged exposure to light, known as photobleaching, can limit the duration of imaging experiments. - Complexity of Sample Preparation: Preparing samples for multicolor imaging can be complex and time-consuming, requiring precise control over the labeling process to ensure specific and non-overlapping signals.
