Molecular Probes™ SlowFade™ Diamond Antifade Mountant with DAPI

Product Code.	Volume (Metric)	unitSize
15312202	2 mL	2mL
15451244	5 x 2 mL	Pack of 5
15471244	10 mL	10mL

Product Code. 15312202 Supplier Molecular Probes™ Supplier No. S36968

Description

SlowFade™ Diamond Antifade Mountant with DAPI

SlowFade™ Diamond Antifade Mountant is a glycerol-based liquid mountant applied directly to fluorescently labeled cells or tissue samples on microscope slides. SlowFade™ Diamond mountant provides superior antifade protection across the entire visual and IR spectrums. You can use SlowFade™ Diamond mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry)to achieve the brightest signal and lowest background.

Features of SlowFade™ Diamond Antifade Mountant include:

Protects fluorescent dyes and fluorescent protein from fading, across entire visible and IR spectrums

Ready-to-use liquid that does not cure; samples can be viewed immediately

Ideal for Alexa Fluor™ and traditional dyes, such as FITC and Cy™3, and fluorescent proteins like GFP, RFP, and mCherry

Maintains fluorescence signal—little to no quenching

Learn more about SlowFadfe™ Antifade mountants.

SlowFade™ Diamond Antifade Mountant comes already mixed and ready to use in a dropper bottle, with or without DAPI. Just apply a drop to the sample, add a coverslip, and image. SlowFade™ Diamond mountant is a non-curing mountant that forms a near-perfect optical path (1.42 RI). When the coverslip edges are sealed with an appropriate coverslip sealant, samples can be stored for days to weeks. If long term storage is desired with a refractive index (RI) closer to immersion oil, then ProLong™ Diamond Antifade Mountant is recommended. With the no-curing-required properties of SlowFade™ Diamond mountant, samples can be evaluated and imaged right after labeling.

Get Photobleaching Protection Across the Entire Visible and IR Spectrums
No matter which fluorescent dyes or fluorescent proteins you are using, SlowFade™ Diamond mountant helps provide the best protection against photobleaching. It is proven to have best-in-class performance against a wide array of fluorescent dyes, including Alexa Fluor™ dyes, FITC, TRTC, Texas Red™, Cy™3, and Cy™5. SlowFade™ Diamond does not quench, and its ability to provide brighter signal, against no background, helps ensure that you obtain the best-quality, high resolution images for publishing.

Stable Signals During Imaging and for Immediate Viewing
SlowFade™ Diamond mountant is the ideal option for analyzing samples right after labeling. Its antifade properties for both fluorescent dyes and fluorescent proteins preserves signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination, or with super resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. A slide sealant can be used to preserve the slides for days or weeks. If longer archiving is desired, then the SlowFade™ Diamond mountant can be washed away with PBS, and the samples remounted with ProLong™ Diamond Antifade Mountant.

Achieve Excellent Refractive Index for Crisp Images
For modern high-resolution imaging applications, refractive index (RI) matching is critical. SlowFade™ Diamond mountant has an RI of 1.42 , which is much closer to the RI of cells or tissue. For thick samples, it might be essential to use SlowFade™ mountaint in order to obtain high resolution images.

Specifications

• Quantity: 2 mL
• Product Type: Antifade Mountant
• Content And Storage: Contains 1 dropper bottle contaning 2 mL reagent.
  
  • Store at 2-8°C or frozen at -5 to -30°C.
  • Protect from light.
  • Stable for at least 6 months after receiving.
• Shipping Condition: Approved for shipment at Room Temperature or on Wet Ice
• Product Line: SlowFade
• Reagent Type: Mounting Solution, Antifade Solution
• Volume (Metric): 2 mL

Frequently Asked Questions (FAQs)

What is the difference between ProLong and SlowFade antifade reagents?

Our ProLong antifade reagents dispense as a liquid that will solidify upon the evaporation of water. SlowFade antifade reagents remain liquid. If you are going to image right away and then dispose of your sample, you do not need a mountant that hardens, such as the SlowFade reagents. If you wish to archive your slide for more than a day, you will want a mounting medium that hardens (or “cures”). This hardening will limit the off-rates of various dye-conjugated antibodies and provides a better refractive index. Also, there will be a lower diffusion rate of free radicals, limiting photobleaching.

Some antifade mounting media stay as liquid whereas others harden. What is the benefit of having one that hardens?

If you are going to image right away and then dispose of your sample, you probably want a mountant that does not harden. If you wish to archive your slide for more than a day, you want a mountant that hardens (or "cures"). This hardening will slow or prevent off-rate of your dye or conjugate and often produces a better refractive index. Secondary sealing is usually not necessary. Also there will be lower diffusion of free radicals, thus limiting photobleaching.

I do not have epoxy or VALAP to seal the coverslip. Do you offer an alternative coverslip sealant?

We offer ProLong Coverslip Sealant (Cat. No. P56128) that can be used to seal the edges of the coverslip and is compatible with both curing and non-curing mountant. The sealant is easy to apply and is brushed on after the mountant has cured, for long preservation of slides. The product page can be found here.

When using a hard-curing mountant, such as ProLong Antifade Mountant, make sure the mountant is fully cured before applying ProLong Coverslip Sealant. Since this sealant can seal moisture under the coverslip, it can interfere with the curing process.

When using a non-curing mountant, such as SlowFade Antifade Mountants, sealing can take place immediately after mounting.

Does Slowfade Antifade Mountant require using a nail polish for sealing?

No. Samples mounted with SlowFade mountants need not be sealed; they are intended for immediate viewing. The coverslip may be anchored (to prevent movement while viewing) by applying molten paraffin to three or four spots around the edge of the coverslip.

I want to mount my dye-labeled cells in an antifade mounting medium to keep the dyes from photobleaching. Which mounting medium do you recommend?

As dyes are illuminated for imaging, they will fade, or “photobleach”, leading to unwanted dimming and lower detection efficiency over time. An antifade mounting medium can greatly reduce photobleaching. If you wish to label live cells, use of ProLong Live Antifade Reagent is helpful. If you wish to mount fixed cells after labeling, and then image immediately and then discard, SlowFade Diamond Antifade Mountant stays liquid but has good refractive index. If you wish to mount your sample and then archive the slides, ProLong Diamond Antifade Mountant will harden to a better refractive index and allow for archiving of the sample for up to several weeks, or even months. Unlike other antifade mounting media, these work well with fluorescent proteins for immediate viewing (archiving fluorescent proteins is not possible), and they are packaged with or without DAPI. More information on these can be found here (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cellular-imaging/fluorescence-microscopy-and-immunofluorescence-if/mounting-medium-antifades/prolong-gold-antifade.html).

My fluorescent dye signal is fading as I image it. What can I do to stop this?

All fluorescent dyes will fade, or “photobleach”, to at least some extent when exposed to strong light at the wavelengths they absorb. Here are some causes for photobleaching and ways to fix the problem:

1) Cause of photobleacing - Generation of free radicals and singlet oxygen
Remedy - i) Use an antifade reagent, which has antioxidants and free radical scavengers:

ii) For live-cell imaging of fluorescent dyes and proteins, we recommend ProLong Live Antifade Reagent which can be added to the cell media or buffer. ProLong Live Antifade Reagent can significantly increase the stability over time for reagents as well as fluorescent proteins, like GFP, without affecting cell health, for up to 24 hours.

iii) For immediate analysis and short-term storage of fixed samples, we recommend SlowFade Diamond Antifade Mountant (it is a liquid mountant and can be used for immediate viewing and then disposal of the sample within a day).

iv) For long-term analysis of Alexa Fluor dyes in fixed samples, we recommend a mountant that hardens, such as ProLong Diamond Antifade Mountant. The harrdening of the mountant also slows diffusion of free radicals).

v) For long-term analysis of all dyes and fluorescent proteins in fixed samples, we recommend ProLong Diamond Antifade Mountant, suitable for archiving slides.

2) Cause of photobleaching - Dye is particularly sensitive to structural modification upon exposure to light.
Remedy - i) Choose a more photostable dye, such as many of our Alexa Fluor dyes.

3) Cause of photobleaching - Intense Illlumination
Remedy - i) Reduce light exposure, for example by reducing laser power or using neutral density filters.

ii) Minimize the viewing time of labeled sample, and close shutter when not viewing.

iii) Use an objective with a lower numerical aperture, such as a lower-power objective.

You can find more information on choosing an antifade reagent on the link below http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cellular-imaging/fluorescence-microscopy-and-immunofluorescence-if/mounting-medium-antifades.html.

DAPI and Hoechst dyes are quite similar to each other. Why would I choose one over the other?

DAPI is a very common blue-fluorescent dye for nuclear counterstaining and gives very bright labeling on nuclei in fixed and permeabilized cells and tissues. However, it is considered to be a semi-permeant to impermeant stain and provides inconsistent staining of live cells. Hoechst 33342 dye is cell-permeant and stains with the same binding mechanism and fluorescent color; it is preferred for live-cell imaging and is just as good as DAPI for fixed cell labeling.

For Research Use Only. Not for use in diagnostic procedures.