The Good, the Bad and the Ugly of 45 color panel design: Building to higher dimensional flow cytometry while maintaining biological resolution

High dimensional full spectrum flow cytometry grants unprecedented access to previously unattainable parameters in cellular biology. Fluorescent dye performance heavily impacts both the quality and the dimensionality of flow cytometry experiments. The development of fluorescent labels engineered with targeted excitation and emission spectra using the Invitrogen™ Phiton™ DNA platform has allowed us to expand the number of biological questions that can be asked in a single panel from 40 to 45 colors. However, as we push the theoretical limit of detection, data analysis increases in complexity with the combination of spectral unmixing and data deconvolution. In this tutorial, we take a deep dive into the data generated by the world's first 45 color flow experiment, examining what works and what doesn't in high dimensional flow cytometry. We will also explore the iterative process involved in developing and evaluating high parameter spectral flow cytometry panels.

 

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