High-Plex Immunocytochemistry Core
The HPIC Core utilizes the Akoya PhenoCycler-Fusion 2.0 to enable high-plex spatial phenotyping of whole tissue slides at single-cell resolution.
Our services include:
• Consultations for project planning and antibody panel design
• High-plex immunocytochemistry workflow from baking and deparaffinization through imaging and data delivery
• Post-staining flow cell removal, H&E, and imaging
• Custom antibody conjugations and optimization for the PhenoCycler-Fusion workflow
Our antibody offerings include:
- 80+ targets for human FFPE tissue
- 60+ targets for mouse FFPE tissue
If you’re interested in using fresh-frozen tissue, please email us.
Core Highlights
In August 2023, the HPIC Core’s work was first featured in The Lancet Oncology. This study used the PhenoCycler-Fusion to characterize tumor immune microenvironment changes in patients from a University of Michigan clinical trial led by Dr. Pedro Lowenstein that utilized a multi-pronged immunotherapy approach for the treatment of glioblastoma. These results demonstrated the power of our workflow, which utilizes conventional tissue slides to map numerous cell types in space. Combining protein expression data with spatial context allows for the exploration of new frontiers in single cell spatial analysis, such as changes in cell neighborhoods and cell-cell interactions between experimental groups.
We have pending publications under review! Pre-prints can be viewed here:
- H3.3-G34R Mutation-Mediated Epigenetic Reprogramming Leads to Enhanced Efficacy of Immune Stimulatory Gene Therapy in Diffuse Hemispheric Gliomas
- A Dual Pronged Approach to Cancer Immune Exclusion: Tumor-Derived LAIR-1 Simultaneously Drives Fibrosis and Blocks Immune Recruitment in Glioma
Our antibody offerings are continuously growing and now include the PhenoCode Discovery IO60 Human Protein Panel and the Mouse Immuno-Oncology PhenoCode Discovery Panel.
Basic Guidelines for Tissue Preparation
– Sections must be between 5 and 10 microns thick
– FFPE tissue must be sectioned on to positively charged slides
– Tissue must be precisely placed within the capture area pictured (left) to be imaged
Our High-Plex Immunocytochemistry Core Researchers
Pedro Lowenstein
Richard C. Schneider Collegiate Professor of Neurosurgery,Professor of Cell and Developmental Biology,
Director of High-Plex Immunocytochemistry Core
Contact Information:[email protected]
The focus of my research program is to discover the cellular, molecular, and neuroanatomical basis underlying the growth patterns of malignant brain tumors, and the interactions between cancer cells with the tumor microenvironment, in both experimental models and in human patients suffering from malignant brain tumors. To do so, we are probing how brain glioma cells migrate throughout the brain and eventually kill the hosts’ neurons and glial cells. Understanding the precise molecular basis of glioma tumor cell growth and invasive behavior will uncover novel therapeutic targets aiming at inactivating the essential mechanisms used by tumors to grow and destroy normal brain tissue and, thus, kill the host. We are studying very early stages of tumor pattern formation using fluorescently labeled glioma cells in combination with advanced in vivo multiphoton imaging technologies. The new information will lead to the development of novel therapeutics for malignant glioma. We have focused on understanding the mechanisms by which the common glioma matrix protein Collagen I modulates tumor and immune function. My lab is also investigating how the interactions of collagen with the specific collagen receptor LAIR-1 modulate glioma malignant behavior, and the complex interactions of the ECM with infiltrating immune cells.
Anzar Abdul Mujeeb
Postdoctoral Fellow, High-Plex Immunocytochemistry Core Contact Information:[email protected]Dr. Anzar Abdul Mujeeb completed his B.S in Biochemistry from AMU, Aligarh, India. He continued his education at Interdisciplinary Biotechnology Unit, AMU, India, first with an M.S in Biotechnology under the supervision of Prof. Asad U Khan, followed by a Ph.D. in Molecular Immunology under the supervision of Prof. M. Owais, investigating the role of cytokines in establishment and survival of intracellular pathogen in the host. He was awarded Research fellowships in Life Science by the Council of Scientific & Industrial Research (CSIR), India, and Dept. of Biotechnology (DBT), Govt. of India, India.
Anzar joined the Castro-Lowenstein team in January 2021 as a Postdoctoral Research Fellow to study the pro and anti-tumor mechanisms mediated by genes involved in autophagy, DNA damage repair (DDR), and immune mechanisms of gliomas. Currently, he is working on neuro immuno-oncology which involves uncovering the role of the mIDH1, CXCR4/CXCL12, and DDR signaling in glioma. His goal is to establish the basis of a new treatment regimen for gliomas based on manipulating the immune, DNA damage repair, and autophagic mechanisms in glioma, leading to novel immunotherapeutic approaches which could be translated into the clinical trials.
Molly West
Research Lab Specialist Senior, Manager of Multiplex Immunocytochemistry Core Contact Information:[email protected]Molly West is the manager of the High-Plex Immunocytochemistry Core and a Research Lab Specialist developing workflows for the acquisition and analysis of multiplex immunofluorescence images from the Akoya PhenoCycler-Fusion. Prior to her work with the Castro-Lowenstein Laboratory, Molly was a senior laboratory technician at the Wyoming State Veterinary Laboratory & University of Wyoming Biocontainment Facility working on pathogen sequencing and genomics. She holds a BS and MS in Biology from University of Michigan and Bowling Green State University, respectively.