Autofluorescence is a well-known problem associated with multiplex imaging applications, including PhenoCycler. We demonstrate a simple autofluorescence reduction method that integrates readily into the PhenoCycler workflow. The protocol improves PhenoCycler imaging data and streamlines antibody panel design by enabling more reliable use of low wavelength fluorophores, such as Alexa 488.

You Will Learn:

• How to set up autofluorescence quenching in the PhenoCycler  workflow
• The effect of autofluorescence reduction on PhenoCycler data and workflow
• Analytical metrics that will assess the effectiveness and safety of this protocol