Abstract: The object of the experiment described in this paper was to demonstrate that cells stained with multiple fluorophores could be identified and quantified simultaneously. Hyperspectral imaging was used to classify spleen cells of a Balb/c mouse, with the anti-mouse CD4 antibody conjugated with Alexa 488, 532, 546 and 568. It was found that the system was able to identify the specific fluorophore present and map their location in the cells. The system also provided relative signal strength data. Spectral libraries were constructed with color-coded spectra that enabled automatic spectral identification in subsequent acquisitions. !3
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