ProLong™ Glass Antifade Mountant with NucBlue™ Stain, 5 x 2 mL - FAQs

View additional product information for ProLong™ Glass Antifade Mountant with NucBlue™ Stain - FAQs (P36981, P36985, P36983)

11 product FAQs found

我该怎样去除混合了DAPI的ProLong Antifade Mountant、ProLong Diamond Antifade Mountant和ProLong Diamond Antifade Mountant抗淬灭封片剂中的气泡？

有以下两种方法来去除气泡：

1.吸取少量你们要使用的ProLong 抗淬灭封片剂（少许过量）于离心管中。盖上盖子并置于离心机中离心（转速7, 000到13,000 rpm）。此时气泡会移动到样品顶部，通过移液枪/枪头即可移去气泡。
2.拧松ProLong 抗淬灭封片剂的瓶盖，但不要打开。将整瓶/管放入真空瓶中，使用抽气龙头（抽气龙头T型管）。打开真空（水随龙头流走）并对混合液抽真空10到20分钟。

为了避免在样品中形成气泡或去除气泡：
1.吸取适量的ProLong 抗淬灭封片剂封片之前可以将吸取体积调高一点。吹打混合液时不要把全部液体都吸上来，枪和枪头向上离开瓶子的时候体积不要满。这样做可以防止气泡进入枪头。
2.使用盖玻片时出现气泡：当把盖玻片放于ProLong 抗淬灭封片剂上封固时，将盖玻片抬起一个小角度后再轻轻放下。如果盖玻片离样品平面过近或是放下过快，都有可能出现气泡。
3.组织中出现气泡：组织切片，尤其是冰冻切片的一个问题是，空气往往会困于切片中或切片下。在封固时没有发现气泡，但是随着封片剂变硬，对样品轻微的挤压就会放出其中的空气。这使得切片中形成部分微小气泡，封于封固剂中。为了避免这种情况，封固之前先进行脱气。将切片浸没在缓冲液或封闭液中，然后放入真空泵上抽真空。这样就会把切片上的空气和缓冲液抽去。将切片从脱气缓冲液中移出然后封固。
4.如果ProLong抗淬灭封片剂封固样品时快要凝固但是出现了气泡，你们可以把玻片放入PBS中（装有PBS的Coplin jar或Petri dish）。ProLong抗淬灭封片剂会膨胀而盖玻片会滑落，或者你们也可以用手轻轻移开盖玻片。你们可以用新的小份的ProLong 抗淬灭封片剂重新封固。

I mounted my antibody-labeled brain cryosection in either ProLong Antifade Mountant, ProLong Gold, ProLong Diamond or ProLong Glass Antifade Mountant. It looked good when first mounted, but after curing, it had tiny bubbles all over the tissue under the coverslip. The sample was air-dried prior to mounting. What happened and what can be done to remove the bubbles?

The tissue likely had air trapped in it, either before the labeling process or as a result of air-drying. While the tissue is in blocking solution or another wash, we recommend putting it in a vacuum to pull out internalized air. Air-drying is not necessary; just tap off excess buffer and mount the damp tissue with the mountant.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What do ProLong Gold, ProLong Diamond and ProLong Glass antifade mountant reagents contain?

They are composed of proprietary antifades and polymers in aqueous buffers.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I use ProLong Gold, ProLong Diamond, or ProLong Glass mounting medium in a multi-well plate?

No. If the sample cannot be sealed with a coverslip, we recommend the use of SlowFade antifade mountant.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What can affect sample curing of ProLong Gold-, ProLong Diamond-, and ProLong Glass-mounted samples?

Our ProLong Gold, ProLong Diamond, and ProLong Glass mounting media harden ('cure') by the evaporation of water. Placing the mounted samples in a humid or refrigerated environment would slow down the evaporation process ('curing').

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What is the concentration of NucBlue Stain in ProLong Glass Antifade Mountant with NucBlue Stain?

The concentration of NucBlue dye in this product is proprietary.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Are ProLong, ProLong Gold, ProLong Diamond, and ProLong Glass reagents compatible with DyLight dye-conjugated antibodies and streptavidin and with other organic dyes?

We have not tested ProLong, ProLong Gold, ProLong Diamond, or ProLong Glass on DyLight dye-conjugated proteins or with other organic dyes, but it is likely that they will be compatible.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I use any of the ProLong, ProLong Gold, ProLong Diamond, or ProLong Glass reagents on live samples?

No. ProLong, ProLong Gold, ProLong Diamond, and ProLong Glass reagents are intended for use on fixed or fixed/permeabilized samples. Only ProLong Live Antifade Reagent (Cat Nos. P36974 and P36975) is intended for use with live cells.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What happens when ProLong, ProLong Gold, ProLong Diamond, or ProLong Glass antifade mountant is curing? Is there a chemical reaction?

The evaporation of water is the “curing” process for Prolong, ProLong Gold, ProLong Diamond, and ProLong Glass antifade mountant. The evaporation of water causes the polymer to harden. There is no chemical reaction. Please note that there is no curing/hardening for any of the SlowFade reagents or ProLong Live products.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

How do I remove bubbles from the vials of either ProLong Antifade Mountant, ProLong Gold, ProLong Diamond or ProLong Glass Antifade Mountants or ProLong-mounted samples?

Bubbles may be removed by one of two methods:

1. Place the amount of ProLong reagent you wish to use on your sample (plus a little excess) in a microcentrifuge tube. Close the cap and centrifuge this aliquot using a tabletop microcentrifuge (speed from 7, 000 to 13,000 rpm). Bubbles should move to the top and these bubbles may be aspirated using a pipettor/pipette tip.
2. Unscrew the lid of the bottle/vial containing the ProLong reagent to make it loose, but do not remove the lid. Place the entire bottle/vial into a vacuum flask, using a faucet aspirator (faucet T-tube). Apply a vacuum (water running through the faucet) and allow vacuum aspiration to occur from 10 to 20 minutes to degas the mixture.

To avoid the formation of bubbles on a sample or to remove bubbles:

1. Before pipetting the desired amount of ProLong reagent for mounting, set the pipettor for a slight excess volume. When pipetting up the mixture, do not pipette up the complete amount, but lift up the pipette tip from the bottle with the pipettor not yet up to full volume. This prevents the aspiration of bubbles into the pipette tip.
2. Bubbles trapped during application of the coverslip: When placing your coverslip onto your drop of ProLong reagent, place the coverslip at a slight angle then, gently lower the coverslip. If the coverslip is lowered flat onto the sample, or lowered too quickly, bubbles can be trapped.
3. Bubbles trapped in tissue: One problem with tissue sections, particularly cryosections is that air can get trapped within and under the section. Upon mounting, bubbles are not observed but as the mountant hardens, it compresses the sample slightly, forcing air out of the section. This leads to microscopic bubbles forming over the section, trapped within the mountant. To avoid this, degas the tissue sample prior to mounting. Place the sections submerged in buffer or blocking solution, into a vacuum chamber and expose the sample to the vacuum. This will degas the sections and buffer. Remove the sample from this degassed buffer and mount.
4. If ProLong-mounted samples have already cured but have bubbles, you can un-mount your sample by placing the slides into PBS (Coplin jar or a Petri dish filled with PBS). The ProLong reagent will swell and the coverslip will slide off or can be gently removed manually. You can then re-mount with a new aliquot of ProLong reagent.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.