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

Immunofluorescence Mountant - Revised 1/2/2006 MAS

1. SOLUTIONS and PROTOCOL:

   10% Phosphate Buffered Airvol-205 (polyvinyl alcohol) with 2% n-propyl gallate.

   Airvol-205 (Air Products and Chemicals Inc., Allentown, PA 18105)

   5.0 g in 20.0 ml n-H2O

2. Disperse Airvol-205 into vortex of water in a 125 ml glass bottle. Allow to hydrate for 15 minutes.
   
   
3. Autoclave for 30 min. to solublize the Airvol-205. Cool to room temperature.
   
   
4. Dissolve 1.0 g n-propyl gallate (Sigma) in 25 ml glycerol (overnight on stir plate).
   
   
5. Mix n-propyl gallate and Airvol.
   
   
6. Add 0.35 ml 0.1M monobasic phosphate buffer.
   
   
7. Adjust pH > 8.0-8.5 with 0.1M dibasic phosphate buffer.
   
   
8. Q.S. with n-H2O if necessary to 50.0 ml.
   
   
9. 1-2 µg/ml DAPI may be added to media to label DNA.
   
   
10. Aliquot and store at -20°C.

    Note:

• pH must be >8.0-8.5. Check solution before every use.
• Centrifuge mountant before use.
• Add small amount to slide (~15-20 µl per 22X50mm cover slip), add cover slip and press gently to remove excess mountant.
• Place at 37°C for 10-15 minutes to polymerize Airvol. The cover slip can still be removed if necessary. Airvol may soften if slides is refrigerated. Cover slip may be sealed with nail polish to make a more permanent seal.