Core D: Tissue Characterization and Immunoreagent Resource

Director: Joanne Murphy-Ullrich, PhD  murphy@uab.edu; (205) 934-0415
Co-directors: 
Gene Siegal gsiegal@uab.edu; Mary Ann Accavitti-Loper maal@uab.edu
Adm Assistant: 
Jane Richard  jrichard@path.uab.edu; (205) 975-9369 

Contacts:
Histology/Morphometry: 
Patricia Lott (205) 934-2007 or (205) 934-4415, LHR B42  
plott@uab.edu
FRET Microscopy: 
Shawn Williams, (205) 934-7403, MCLM 684A, tzaron@uab.edu
ERIC facility
Mary Ann Accavitti-Loper, PhD, (205) 934-0913, VH219,  maal@uab.edu

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 

Core Services ( Contact Director for protocol details)

Resources
Cost
Histology/Morphometry Services
contact Ms. Lott
   
FRET Microscopy and Imaging
$15.00/hr after 1st hour of raining
   
Epitope Recognition and Immune Reagent Core
 
Immunization (2 mice)
$200.00
1 rat
$250.00
Total Fusion Cost
$2000.00
   
Electron Microscopy Services
 
Transmission Electron Microscopy--- FEI Tecnai T12 Spirit/Assisted $40.00/hr
Transmission Electron Microscopy--- FEI Tecnai T12 Spirit/Unassisted $30.00/hr
Sample Processing per sample $60.00/sample
Ultra-thin and semi-thin sectioning per sample with post stain $25.00/hr
Negative Staining per Grid $10.00/Grid
Compact Disc $2.00 each
High Resolution Printing: Inkjet, $1.50/ print $1.50 each
Services Order Form  
   
Other Services
 
Subcloning per line
 
Vial Storage
$0.50/vial/year
Ascites Production (subject to UAB IACUC guidelines)
 
2 BALB/C Mice (typical yield 5 to 10 ml)
$200.00
-2 Nude or RAG2 mice
$250.00
Large-scale in vitro mAb Production, 500 ml IgG-free media
$350.00
Large-scale in vitro mAb Production, 1000 ml IgG-free media
$500.00
Protein G purification from supernatant fluid or ascites
$200.00
Phage Library Screening with work-up of 10 positive clones
$2000.00
Stock Monoclonal Reagents (Contact Core for pricing and availability  (205) 934-7156