Does anyone have any experience using multiplexed IHC methods in fresh frozen human tissue, ideally brain? We are currently trying to set up techniques for labelling of a-synuclein Lewy Body Pathology and Neuronal cells in fresh frozen human brain and although we've had some limited success with a protocol utilising an IHC DAB imaging for synuclein, the background we are getting from non-IHC based methods of neuronal counterstain (cresyl violet, H&E) is very high and making assessment difficult. These slides will be used for Laser Capture, so we are limited in what we are able to do in processing the slide. Ideally we want to remain with chromagenic detection and not use immunoflorescence.
We are interested in exploring a multiplexed IHC platform, such as the Immpress Duet Double staining HRP/AP Polymer Kit that vector provide. Does anyone have any experience with this kit or recommendation of other multiplex platforms for dual IHC staining in fresh frozen, unfixed human tissue (ideally brain).
Please go to this link :
https://www.atlasantibodies.com/resources/antibody-applications/protocols/ihc-if-protocol-multiplex/
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