USER NOTES
IM037-100
FluoroGel with Para Phenylenediamine (PPD) Anti fading Mounting Medium
Description:
Fluoroshield with strong antifading agent, 1,4-phenylenediamine (PPD) is an aqueous mounting medium for preserving fluorescence of tissue and cell smears. This unique formula prevents rapid photobleaching of FITC, Texas Red, AMCA, Alexa fluoro 488, Alexa fluoro 594, tetramethyly rhodamine, and Redox. The fluorescence is retained during prolonged storage at 4°C in the dark. This medium contains phenylenediamine and is not suitable for immunofluorescence of Cy dyes, GFP, mCherry, Phycoerythrin (R-PE), phyocyanin (PC), and allophycocyanin (APC) and other fluorochromes proteins.
Refractive Index
1.366 ± 0.002 (This number applies to this mounting medium in solution. Refractive indexes change when the water solvent evaporates and mounting media dries on slides. We do not have the means to measure the refractive indexes of dry mounting mediums; however, we expect the numbers to go higher when dried. The refractive index of water is 1.3330)
May encounter problems with frozen brain or other frozen tissues with lots of fat.
Intended Use
Immunofluorescence, confocal microscopy.
Reagent
Ready to use mounting medium with dark coffee colour. This colour does not interfere with Immunofluorescence.
Storage
2-8°C is recommended. 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 open the dropper to remove any air bubbles.
Apply 2-3 drops of mounting medium directly on top of the specimen.
Let stand at room temperature for about 3-5 minutes in the dark.
Apply cover slip carefully avoiding air bubbles.
The specimen is ready for visualization under a microscope.
One can seal the edges of cover slip with nail polish, any organic medium. If a coverslip is not sealed air bubbles will appear in few days.
Method for applying Coverslip: Put 1-2 drops of Mounting medium 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 37°C for one hour in the dark to dry organic mounting medium.
For 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) distilled or deionized water for several minutes. Carefully and slowly move the coverslip. Soak in water for an additional few minutes to remove coverslip. Rinse slides several times with warm water to remove all mounting medium. The slide can be remounted again.
Warning
For research use only; not for use in diagnostic procedures. FOR IN VITRO LABORATORY USE ONLY
“In vitro laboratory products for research”
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