Cell Biology & Imaging Core

The objectives of the Cell Biology and Imaging Core are to provide high quality histopathologic and scientific support for research projects by localizing proteins and/or mRNAs in tissues and cultured cells, and to offer pathology support. The Core maintains a library of wild-type murine embryos and adult tissues embedded in paraffin for use by O'Brien Center investigators. The lab also maintains bright-field, dark-field, phase-contrast, epifluorescence, and confocal microscopes.

Services

  • Pathology
    • Processing of tissues for routine paraffin embedding
    • Detailed paraffin and cryo-microtomy
    • Standard histologic stains and special stains when required
    • Immunohistochemistry and in situ hybridization
    • Kidney pathology
    • Single tubule isolation and immunocytochemistry
    • Laser microdissection
  • Cell Biology
    • Immunocytochemistry
    • Light and confocal microscopy and image analysis
    • Live cell imaging
  • Serum Assays
    • Soluble Klotho
    • Fibroblast growth factor 23
  • Magnetic resonance-based imaging
    • Imaging of rodent kidney and heart including functional MRI
      • Apparent diffusion coefficient (fibrosis)
      • Arterial spin labeling (blood flow)
      • BOLD (oxygenation) Na MRS

IP-IB Klotho Assay

This assay is provided by the O'Brien Center as a service that incurs non-profit cost-recovery charges. In contrast to the commercial ELISA, we use an immunoprecipitation-immunblot assay using synthetic anti-Klotho antibodies that reliably detects full-length Klotho in sera and urine from human, rat, and mouse.

Compared to the commercial assay, the drawback of our IP-IB assay that it requires higher sample volume and cannot achieve high throughput mode, because of the laborious nature of the assay. However, the main advantage is the highly specific and sensitive reagents and known performance of inter-assay variation of 7 percent and intra-assay variation of < 2 percent.

Mode of Operation

  • Investigators and their key personnel meet with Core Directors and staff before launching a project.
  • The Core Directors are actively involved in the experimental design of the various protocols.
  • The investigators determine the level of involvement from their laboratory. The Core may provide all the service or the investigator can work collaboratively with the Core.
  • A mutually agreeable plan is formulated, and the estimated cost recovery and time line is presented to the investigator.

Stained tissue slices