Search
Search
View additional product information for SlowFade™ Diamond Antifade Mountant - FAQs (S36972, S36963, S36967)
8 product FAQs found
染料需要发光成像，在此过程中它们会淬灭或“光漂白”，导致荧光不断变暗、检测效率不断降低。抗淬灭封固剂可以大幅减少光漂白现象。如果您想标记活细胞，可使用ProLong Live抗淬灭试剂。如果您想标记后立即封固细胞，然后立即成像并丢弃样品，可采用SlowFade Diamond 抗淬灭封固剂，因为其可以保持液态并有良好的折射率。如果您想封固样品后存档样品，ProLong Diamond 抗淬灭封固剂会固化以获得更好的折射率，且可存档样品长达数周乃至数月。与其他抗淬灭封固剂不同，此类产品非常适合对荧光蛋白质进行即时成像（无法进行荧光蛋白质存档），并且有含或不含DAPI两种包装规格。更多信息请见此处。
当它们暴露在吸收波长强光下时，所有的荧光染料都会在一定程度上出现褪色或“光漂白”现象。这里列有一些光漂白的原因以及解决方案：
1）光漂白的原因：自由基和单线态氧的产生
可能的解决方案：使用具有抗氧化剂和自由基清除剂的抗褪色剂：
i. 对于荧光染料和蛋白质的活细胞成像，我们建议使用ProLong Live Antifade Reagent，该试剂可以加入到细胞培养基或缓冲液中。ProLong Live Antifade Reagent可以在24小时内不影响细胞的健康的前提下显著增加试剂和荧光蛋白，如GFP的稳定性。
ii. 对于直接分析与短期储存的固定样品，我们建议使用SlowFade Diamond Antifade Mountant（该试剂保持液态并能即看即用，然后在一天内丢弃样品）。
iii. 对于长期分析Alexa Fluor染料的固定样品，我们推荐固化的封固剂如ProLong Diamond Antifade Mountant（其可减慢自由基运动）。
iv. -对于长期分析所有染料和荧光蛋白的固定样品，我们推荐ProLong Diamond Antifade Mountant（固化后可以保存载玻片）。
2) 光漂白的原因：染料对褪色尤为敏感
可能的解决方案：
i. 选择更耐光染料，比如我们的Alexa Fluor染料
ii.您可以使用复染剂选择和设置像场，然后切换到目标染色加以成像。
3) 光漂白的原因：强光照射
可能的解决方案：
i. 减少曝光，例如降低激光功率或使用中性密度滤光片。
ii. 观察标记样品的时间尽可能的短，并在不观看时关闭遮板。
iii. 使用具有较低数值孔径的物镜，如低功耗物镜。
这里（https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cellular-imaging/fluorescence-microscopy-and-immunofluorescence-if/mounting-medium-antifades.html?icid=fr-antifade-main）是一个很好的抗淬灭剂选择指南。
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.
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).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.