. Author manuscript; available in PMC: 2016 Apr 1.

Published in final edited form as: Cell Tissue Res. 2015 Jan 27;360(1):71–86. doi: 10.1007/s00441-014-2087-2

Table 3.

Key differences in QD-immunohistochemistry protocol.

IHC Workflow	QD-IHC Specific
Sample Preparation	Works well with fresh, frozen or formalin-fixed, paraffin embedded samples
Deparaffinization/Rehydration	Toluene recommended (replacing xylene^*)
Decloaking of antigens	Same as traditional temperature, pressure, pH and time of cycles is crucial to retrieval of epitopes
Blocking and Antibody incubation	Hydrophilic barrier pens such as ImmEdge by Vector^* (Xing et al 2007)
Dehydration and mounting	Toluene recommended (replacing xylene^*) - both for rinses and mounting media
Microscopy	Spectral camera and software recommended (e.g. Nuance)
Analysis	Spectral unmixing; generation of spectral fingerprints or comparisons of label intensity

An important underlying theme in the difference between traditional and QD-IHC protocols is to utilize solvents such as toluene, chloroform and hexane to prevent interaction with QD surface chemistry that may lead to quenching.