Fluoroshield™ with PI

1. Bring the vial to room temperature. 2. Rinse slide to be mounted with DISTILLED OR DEIONIZED WATER, touch the edges of slide on a paper towel to remove excess water. Place slides on a flat surface away from light. 3. Turn the vial upside down and remove any air bubbles. 4. Apply 2-3 drops directly of mounting medium on top of the specimen. 5. Let stand at room temperature for about 3-5 minutes in the dark. 6. Apply cover slip, carefully avoiding air bubbles. 7. The specimen is ready for visualization under microscope. 8. One can seal the edges of cover slip with nail polish or any organic medium or our Organo mounting medium (cat. # AR-6504). If coverslip is not sealed air bubbles will appear in few days. 9. Method for applying Coverslip: Put 1-2 drops of FLS on the specimen. After 3-4 minutes apply coverslip carefully avoiding air bubbles. Put Kimwipes on the top of coverslip, press gently to remove excess mounting medium. With 200 micropipette add organic mounting medium to seal the edges. Incubate at 37Cfor one hour in the dark to dry organic mounting medium. 10. For long term storage it is recommended that the slide be stored in the dark at 2-8oC.

Fluoroshield mounting medium with PI is an aqueous mounting medium for preserving fluorescence of tissue and cell smears. This unique formula prevents rapid photobleaching of FITC, Texas Red, AMCA, Cy2, Cy3, Cy5, Alexa fluoro 488, Alexa fluoro 594, Green fluorescent protein (GFP), tetramethyly rhodamine, Redox, Phycoerythrin (RP-E), Phyocyanin (PC), and Allophycocyanin (APC). The fluorescence is retained during prolonged storage at 4oC in the dark. This medium does not contain phenylenediamine, which destroys immunofluorescence of Cy dyes, RP-E, PC and APC. This mounting medium is fortified with PI which is a counter-stain for DNA. This product is to be used in in situ hybridization techniques or other methods where fluorescence of DNA staining is required. PI excites at 535nmand emits at 615nm, producing a red fluorescence. RNA is also stained with PI. May encounter problems with FROZEN BRAIN OR OTHER FROZEN TISSUES WITH LOTS OF FAT