Fluoroshield™ with DAPI, histology mounting medium
liquid
Sodium Azide, ≤1% (also contains other ingredients)
Tris-HCl
immunofluorescence: suitable
7.9-8.3
water: soluble
λex 360 nm; λem 460 nm
diagnostic assay manufacturing
hematology
histology
2-8°C
10 - Combustible liquids
WGK 3
Not applicable
Not applicable
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What could be the reason for the occurrence of a white film after curing, particularly when the coverslips are slightly wet with PBS?
The following recommendations are provided:
•If there is tissue attached to glass slides, it is recommended to use a Kimwipe to carefully dry the slide around the tissue section.
•If cells are on a coverslip, removing excess water from the coverslip by placing it on its end and then using a paper towel to wick away excess moisture is suggested.
•The possibility that as the slides dry, salts from the PBS are evaporating and leaving a white residue or film is considered.
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How is Fluoroshield (F6057) used with DAPI?
Fluoroshield with DAPI is an aqueous mounting medium designed to preserve the fluorescence of tissue and cell smears. This unique formula prevents rapid photobleaching of various fluorescent dyes. Additionally, it retains fluorescence during prolonged storage at 4°C in the dark. Importantly, the medium does not contain phenylenediamine, which can destroy immunofluorescence of certain dyes. The mounting medium is fortified with DAPI, a counter-stain for DNA, intended for use in situ hybridization techniques or other methods requiring fluorescence of DNA staining. DAPI excites at 360nm and emits at 460nm, producing a blue fluorescence. RNA is also stained with DAPI. The intended use of this product includes immunofluorescence and confocal microscopy.
Reagent:
Fluoroshield with DAPI is a ready-to-use mounting medium.
Storage:
It should be stored at 2-8°C, protected from light, and should not be frozen.
Procedure:
- Bring the vial to room temperature.
- Rinse the slide to be mounted with distilled or deionized water, remove excess water by touching the edges of the slide on a paper towel, and place the slides on a flat surface away from light.
- Apply 3-4 drops of mounting medium directly on top of the specimen and spread it out evenly, ensuring the tissue is not touched. Excess medium can be removed by touching the edges of the slide on a paper towel.
- Let it stand at room temperature for about 5 minutes.
- Apply a cover slip, carefully avoiding air bubbles. If a coverslip is not used, air bubbles will appear in a few days.
- For long-term storage, it is recommended to store the slide in the dark at 2-8°C.
Removal of Coverslip:
The coverslip can be removed before sealing the edges. Soak the slide in warm (37°C) water for a few minutes, carefully and slowly moving.
Helpful?
What are the procedures for solubilizing and using Fluoroshield with DAPI (F6057)?
Fluoroshield with DAPI is a solution that is ready to use and does not require solubilization. It is an aqueous mounting medium designed to preserve the fluorescence of tissue and cell smears. The mounting medium is fortified with DAPI, a counter-stain for DNA, and is intended for use in situ hybridization techniques, immunofluorescence and confocal microscopy.
Helpful?
Is sealing a 96-well plate necessary in the final step of ICC with Fluoroshield™ with DAPI, as done with coverslips using clear nail polish?
Fluoroshield with DAPI is designed for use with glass slides or coverslips intended for analysis using a fluorescent microscope. It is not commonly used for coverslipping 96 well plates. There are no known individual, small, round coverslips for each of the 96 wells, nor are there large clear glass plates to fit over the top of a 96 well plate. If such glass plates are available, it is not known that such a product exists.
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What are the potential implications of storing a bottle of F6057-20 at -20°C for 2 days by mistake?
It is suggested to allow the bottle to thaw. Once thawed, the bottle should be inverted several times to thoroughly mix its contents. If the solution looks normal and does not contain any precipitate or particulate matter, it is expected that the mounting media is suitable for use.
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The refractive index is not determined internally for this product. However, the supplier reports that it is 1.364 ± 0.002 of the product in solution.
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This product has been used for cell imaging. Please see the following protocols and review the pee-reviewed articles:
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How does the storage temperature relate to shipping conditions?
The storage conditions that a Sigma-Aldrich catalog and label recommend for products are deliberately conservative. For many products, long-term storage at low temperatures will increase the time during which they are expected to remain in specification and therefore are labeled accordingly. Where short-term storage, shipping time frame, or exposure to conditions other than those recommended for long-term storage will not affect product quality, Sigma-Aldrich will ship at ambient temperature. The products sensitive to short-term exposure to conditions other than their recommended long-term storage are shipped on wet or dry ice. Ambient temperature shipping helps to control shipping costs for our customers. At any time, our customers can request wet- or dry-ice shipment, but the special handling is at customer expense if our product history indicates that the product is stable for regular shipment.
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What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
Helpful?
How do I use Fluoroshield (Product Nos. F6182, F5932, F6057)?
1. Bring the vial to room temperature. 2. Rinse slide to be mounted with DISTILLED OR DEIONIZED WATER, then 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 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 0.2 ml plastic pipette tip, making sure the tissue is not touched. Excess medium can be removed by touching the edges of slide on paper towel. 5. Let stand at room temperature for about 5 minutes. 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. If coverslip is not used air bubbles will appear in few days. 9. For long term storage it is recommended that the slide be stored in the dark at 2-8°C.
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