SlowFade™ Gold Antifade Mountant with DAPI, 5 x 2 mL - FAQs

DAPI是一种非常普通的蓝色核复染荧光染料，能对固定和通透的细胞和组织的细胞核进行非常明亮的标记。遗憾的是，人们普遍认为它是介于半通透性到非通透性之间的染色剂，对活细胞的染色效果也不一致。Hoechst 33342染料是细胞通透性染料，与DAPI染色有相似的染色结合机制和荧光颜色；它是活细胞成像的首选，且对固定细胞的标记效果和DAPI一样好。

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.

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

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.

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

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.

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.

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

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.

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