Description
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), tetramethyl rhodamine, Redox, Phycoerythrin (RP-E), Phyocyanin (PC), and Allophycocyanin (APC). The fluorescence is retained during prolonged storage at 4°C 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 hybridisation techniques or other methods where fluorescence of DNA staining is required. PI excites at 535nm and emits at 615nm, producing a red fluorescence. RNA is also stained with PI.
Intented use
Immunofluorescence, confocal microscopy.
Reagent
Ready to use mounting medium.
Storage
2-8°C, protect from light, DO NOT FREEZE.
Procedure
- Bring the vial to room temperature.
- 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.
- Turn the vial upside down and remove any air bubbles.
- Apply 3-4 drops directly of mounting medium on top of the specimen and spread out evenly by tilting the slide back and forth or spread evenly with a 0.2ml plastic pipette tip, making sure the tissue is not touched. Excess medium can be removed by touching the edges of slide on a paper towel.
- Let stand at room temperature for about 5 minutes.
- Apply cover slip, carefully avoiding air bubbles.
- The specimen is ready for visualisation under microscope.
- One can seal the edges of cover slip with nail polish or any organic medium or Organo Mounting Medium. If coverslip is not used air bubbles will appear in a few days. 9. For long-term storage it is recommended that the slide be stored in the dark at 2-8°C.
Removal of coverslip
?Coverslip can be removed before sealing the edges. Soak slide in warm (37°C) water for a few minutes. Carefully and slowly move the coverslip. Soak in water an additional few minutes and remove coverslip. Rinse slide several times with warm water to remove all mounting medium. The slide can be remounted.