Figure 1.

Schematic illustration of hyperspectral imaging of multicolor QD-labeled tissue specimens. Conventional RGB cameras display images in three color channels (red, green, and blue) and thus cannot distinguish spectrally overlapping fluorophores. In contrast, hyperspectral imaging works in a way similar to spectroscopy in that it samples the emission spectra of every pixel at a series of wavelengths (thus creating an image cube). For example, blue QDs only appear in the images taken at shorter wavelength, whereas red QDs appear at longer wavelengths. Based on the spectral information obtained for each pixel, the fluorescent components (multicolor QDs and autofluorescence) can be unmixed into separate images for quantification or merged together with the autofluorescence removed. The use of QDs and hyperspectral imaging enhances the ability of each technique in multicolor imaging.