Immunology

The Immunology Core Module (IM) provides a wide range of support and assays to assist with priority, NEI R01 funded PIs in their research in assessing the immunological basis of eye diseases. We strive to assist investigators with study design, sample processing, protocol development, need based assay development, data analysis, and development of infection/inflammation related animal models. Training is required to enable investigators and their staff to use the flow cytometer.

PIs and their lab members are encouraged to contact the module manager prior to planning any experiment (that requires use of this module). A review of protocol design, selection of correct fluorophore and antibody is necessary for instrument utilization. 

Services provided

• Analysis of Intracellular cytokine expression in permeabilized cells
• Apoptosis assays
• Bone marrow chimera development
• Cytek Northern Lights full Spectrum Flow cytometer
• DNA/Cell cycle analysis
• ELISPOT assays for cell associated cytokine production
• ELISA
• Evaluation of cytokines and chemokines from aqueous and vitreous humor, corneal and retinal lysates and tissue culture supernatants by ELISA
• Flow Cytometry services (Accuri 6C and software)
• Flow cytometric functional assays, including NK cell function, antigen-specific CD4+ and CD8+ T cell cytokine expression, degranulation and proliferation. Evaluation of cell signaling by Phosflow using monoclonal antibodies specific for phosphorylated proteins
• Flow Cytometer Based Cytokine Bead Array
• Immunophenotyping of whole blood, PBMC, and aqueous and vitreous fluids for: NK, DCs, Macrophages, B and T cell percentages, and their subsets, including maturation, trafficking, senescence, activation, in vivo proliferation and chemokine receptor expression
• Isolation of peripheral blood mononuclear cells (PBMC) for staining, cell sorting, or viable cryopreservation
• Isolation of mucosal mononuclear cells (MMC) from mucosal fissure biopsies
• Morphological and biochemical assays for antigen-induced cell death
• Western Blot
• Northern Lights Flow cytometer

How to utilize

Please contact Robert Wright to utilize the core services and for protocol development. Please complete the work order and submit to core manager.

Pricing information

• Flow Instrument Usage (Assisted Use: $40/hour,  Unassisted use for trained users: $25/hour)  
• Data Analysis: (Assisted $35/hour, Unassisted for trained users: $20/hour)
• New immunoassay development: $30/hour
• Cell Sorting (User must provide cartridge with tentative is $200, please contact Robert about ordering) - Assisted Use: $40/hour
• Western: $200/cartridge 
• ELISA: No charge, kit and reagents must be provided by the user
• BMDM: No charge, user must provide mouse and reagents

Dr. Ashok Kumar, Ph.D.
Module Director
Associate Professor - Department of Ophthalmology, Visual and Anatomical Sciences
Kresge Eye Institute
4717 St Antoine RM # K416
Phone: 313-577- 6213

Robert Wright
IM Module Manager
Kresge Eye Institute
4717 St Antoine RM # K435
Phone: 313-577-6202 | 313-577-6213 | 517-304-5158