About the Center

Location and Directions

Data Management System

Contact Information:

Director
Mark A. Sanders

Staff
Gilbert G. Ahlstrand
Tracy E. Anderson
Gail Celio

Students
Chemi Banari
Kaitlyn Bissonnette
Alex Cramer
Lizzy Watkins

University of Minnesota
Imaging Center
Rooms 23-35 Snyder Hall
1475 Gortner Avenue.
University of Minnesota,
St. Paul, MN 55108

Hours: 9:00-5:00 weekdays
Closed holidays

Immunolabelling of LR White / Gold Sections

1. Prepare clean labeling surface by laying down a long piece of parafilm on a wet bench and pressing it down to make it adhere tightly.
   
   
2. Quench unreacted aldehyde groups by placing grids on drops of 0.1M NH₄Cl in PBS for 10 minutes @ RT.
   
   
3. Block non-specific binding sites by transferring grids on drops of 1% fish skin gelatin in PBS, for 20 minutes @ RT.
   
   
4. Prepare moist chamber using large Petri dish and pieces of filter paper soaked with water. Incubate grids on 10µl drops of primary antibody diluted in PBS/1% FSG for 30 min @ RT.
   
   
5. Wash 4 x in PBS (5 minutes total).
   
   
6. Label grids with secondary antibody (if necessary) for 30 minutes @ RT.
   
   
7. Wash 4 x in PBS (5 minutes total).
   
   
8. Label grids with Protein A Gold for 30 min @ RT.
   
   
9. Wash 4 x in PBS (5 minutes total).
   
   
10. Fix in 1% glutaraldehyde in PBS for 5 min.
    
    
11. Wash grids 5 x with distilled water. Air dry.
    
    
12. Stain grids with 2% uranyl acetate and lead citrate.