Are you interested in quantitating immune cells that infiltrate the human tissues you study, understanding where they are located, or determining which other cells they are near in the tissue architecture? HIMSR operates a state-of-the-art imaging facility for many of your imaging needs. We are equipped with Akoya Biosciences’ Vectra Polaris, an automated microscope that images brightfield or immunofluorescence slides. We also offer IONpath’s MIBI (Multiplexed Ion Bean Imaging) technology. We have pre-developed antibody panels for Vectra and MIBI or can help develop new antibodies to help you analyze your tissues.
These technologies are optimal for analysis of formalin-fixed, paraffin-embedded tissues or tissue microarrays (not frozen tissues). The antibodies are resolved with Akoya Opal technology, which allows use of the best primary antibodies for immunohistochemistry, regardless of species with no cross-reactivity. Log into iLabs for fees; contact Angie Minic for a quote.
With this high-throughput instrument, we can distinguish 7 colors in a full-slide scan or 9 colors in regions-of-interest using up to a 40x objective. We have 10 standardized panels to choose from that are free of development charge (see list below and poster). Alternatively, we can custom build antibody panels from over 350 different human and mouse antibodies. For every antibody panel, we include stained tonsil-tissue controls, scan slides with up to a 40x objective, and can provide analysis depending on your needs.
Basic Phenotyping Panel 1 – CD4, CD8, CD20, FOXP3, CD45RO, CK, DAPI
Basic Phenotyping Panel 2 – PDL1, CD19, CD3, CD68, CD8, CK, DAPI
Myeloid Panel 1 – CD33, PDL1, CD11C, HLADR, CD68, CD8, CK, DAPI
Myeloid Panel 2 – PDL1, HLADR, CD163, CD14, CD68, CK, DAPI
Structural Myeloid Panel – SMA, CD31, CD64, FAP, CD14, CD206, DAPI
Basic Phenotyping Panel – CD3, CD4, CD19, CD56, CD8, FOXP3, CD68, CK, DAPI
Myeloid Panel – CD33, CD15, CD14, CD11B, CD56, CD68, CK HLADR, DAPI
Immune Checkpoint Panel 1 – CD3, CD19, Granzyme B, PDL1, Epcam, PD1, CD68, Ki67, DAPI
B-cell Panel – CD3, IgD, CD27, CD138, CD20, CD163, CD68, CK, DAPI
Immune Checkpoint Panel 2 – SMA, CD31, CD64, FAP, CD14, CD206, DAPI
Opal technology includes 8 fluorophors (for Vectra Polaris) and enhanced DAPI to stain DNA which are read at different wavelengths. Briefly, we apply a primary detecting antibody, a secondary antibody, the Opal reagent, microwave to remove the antibodies but not the Opal reagent, repeat.
inForm is a patented image analysis software package that uses trainable algorithms in a user-friendly interface to accurately visualize and quantify biomarkers in tissue sections. Capability includes phenotyping, co-localization, co-expression, tissue region detection (e.g. tumor versus stroma), and traditional scoring functions. This software is available for use on a dedicated analysis station within the HIMSR (sign up for time on the iLab calendar) or, for larger projects, check out a copy of inform from the HIMSR ($). We offer periodic workshops to help you get started with inForm and Phenoptr Reports and Akoya Biosciences’ also have great new videos on how to use these softwares [inForm and Phenoptr Reports].
Training Workshop Video - UCD login required to watch. (If you are having trouble, try opening a new incognito tab and pasting the link there.)
Using the Leica Autostainer, we have developed standardized antibody panels for the analysis of tissue microenvironments in human FFPE tissue for detection by the Vectra instrument. The Leica Autostainer can perform the entire 7-color protocol on 30 slides in one overnight batch; 9-color protocols take 1.5 days. We can also run smaller batches of slides or multiple different panels at one time. If you wish to perform the staining in your own laboratory, we are happy to provide the antibody clone names and staining protocols.
Custom IF antibody panel
If your antibody of choice is not yet available, we can work with you to develop a custom antibody panel either in your lab or HIMSR. In the spirit of fostering collaborative research at CU|AMC, antibody panels developed in HIMSR will be shared with the campus as future standard panels.