Analysis of genetically engineered mouse models of PKD will require thorough morphological examination of tissues. The objective of the Tissue Characterization and Immunoreagent Resource (TCIR) is to provide P30 investigators with access to sophisticated, state-of-the-art technology for the detection and characterization of morphological changes associated with mouse models of recessive PKD. These services include: Histology [routine, plastic]; immunohistochemistry; immunofluorescence; Transmission electron microscopy; animal necropsy; morphometry; pathology evaluation; and limited molecular analyses (laser capture microdissection, in-situ hybridization). The Core will also provide user support and instrumentation for live cell FRET microscopy, fluorescence, brightfield, and phase microscopy imaging, x-y axes particle tracking. Another function of the TCIR is to supply investigators with expertise in monoclonal antibody (mAb) technology. Services offered by the mAb unit include; design and analysis of peptide antigens, creation of new hybridoma lines, large-scale production and purification of mAbs and development of antigen-specific human ScFvs. In addition, commonly used monoclonal reagents (i.e. anti-HIS, anti-MYC, anti-GST) will be available at minimal cost.

The Electron Microscopy Laboratory is affiliated with the High Resolution Imaging Facility at UAB and is a full service laboratory providing access to electron microscopes and equipment for sample preparation. In addition, the facility is staffed by a skilled Research Associate who can assist users with the experimental design, sample processing, and image acquisition and analysis. The services provided include assistance with both traditional methods and more recent technical developments to suit the investigator's needs. The services that are available are as follows: 

Transmission Electron Microscopy (TEM):

1. Standard routine processing of specimens with fixation through embedding.
2. Standard thick and thin sectioning with staining.
3. Viewing and photography on the TEM.
4. Digital images provided on CD or memory stick/flash drive which user provides. Standard supplies (including some fixative) are provided.

Routine processing includes either tissue or cell cultures either as monolayers, on membranes, or as pellets with special orientation. The facility has the capability to do negative staining.

These procedures are provided as a full service. If an investigator has sufficient knowledge and ability for use of the equipment then the facility allows for independent work at a lower cost, see fee schedule. Training can be provided.

Immunocytochemistry for TEM:

1. Standard routine processing of specimens with fixation and embedding, in either LR White or LR Gold
2. Standard routine thick and thin sectioning for post-embedding immunostaining*
3. Immunostaining*
4. Viewing and image acquisition on TEM

*Antibodies provided by the investigator, pre-embedding immunostaining can be done if desired.

Equipment Available in the EM Center:

TEM Microscope: Tecnai Spirit Twin 20-120kv (FEI, Hillsboro, OR), new in the fall of 2006
Ultramicrotome: Reichert Ultracut E