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Try our upgraded product VECTASHIELD® PLUS Antifade Mounting Medium with DAPI
VECTASHIELD Antifade Mounting Medium with DAPI is a unique, stable formula for preserving fluorescence. This medium inhibits rapid photobleaching of fluorescent proteins and fluorescent dyes. VECTASHIELD Mounting Medium with DAPI does not solidify, but remains a liquid on the slide and can be stored without sealing.
| Unit Size | 10 ml |
|---|---|
| Applications | Immunofluorescence, In situ hybridization, Cellular Imaging |
| Mounting | Aqueous (Non-Hardening) |
| Antifade | Yes |
| Counterstain | DAPI (blue fluorescence) |
| Cellular Stain | Nucleus |
Of the many variations of super resolution microscopy (SRM) currently being used, investigators have found the properties of product H-1000 (VECTASHIELD), to be advantageous in stochastic optical reconstruction microscopy (STORM) and structured illumination microscopy (SIM). Refer to the following published references for further information: 1) Olivier, N., et al (2013) Simple buffers for 3D STORM microscopy. Biochemical Optics Express 4:885-899. 2) Wegel., et al (2016) Imaging cellular structuires in super-resolution with SIM, STED and localisation microscopy: A practical comparison. Scientific Reports 6:27290.
All of the VECTASHIELD formats we currently offer which includes regular non-setting VECTASHIELD, PLUS, HardSet and Vibrance formulations each contain a percentage of glycerol.
No form of tissue dehydration (e.g. air drying or ethanol exposure) is required nor recommended when applying VECTASHIELD. From our experience, the most optimal antifade actions of VECTASHIELD are obtained when the preparation is removed from the final buffer/water rinse, kept slightly wet/moist and then coverslipped with a small volume (25-50 uL) of VECTASHIELD.
That depends upon how long you wish to retain the slides and which VECTASHIELD formulation you are using. If you are using one of our non-setting VECTASHIELD products such as H-1000/H-1200 or H-1900/H-2000, then we suggest sealing the coverslip with plastic sealant or nail polish if you intend to keep the slides beyond a day or so. If you are using one of our setting/curing formulations such as VECTASHIELD HardSet or Vibrance, then in most cases when using thin cut (<10 um) tissue sections or cell monolayers.
VECTASHIELD Mounting Media are compatible with a wide array of fluorochromes, enzymatic substrates, and fluorescent proteins. For more information about the compatibility of VECTASHIELD products, please consult the VECTASHIELD® Mounting Medium Compatibility table.
All formulations of VECTASHIELD Antifade Mounting Media are available with or without the counterstain DAPI (4′, 6-diamidino-2-phenylindole), which produces a blue fluorescence when bound to DNA with excitation at about 360 nm and emission at 460 nm. If desired, the concentration of DAPI can be reduced by mixing with the corresponding VECTASHIELD Mounting Medium (without DAPI).
If desired, coverslips can be sealed around the perimeter with nail polish or a plastic sealant. Mounted slides should be stored at 4 °C, protected from light.
The refractive index for VECTASHIELD Mounting Medium is 1.45.
Other manufacturers measure the antifade properties of their mountants using labeled microspheres or arrayed spots. Vector Labs prefers to measure antifade properties of VECTASHIELD mountants using frozen tissue sections immunohistochemically stained with fluorescently labeled secondary antibodies. Antifade capability is then measured using a 40x objective with real time imaging over 30 seconds of continuous exposure to the excitation illumination. Individual intensity measurements are recorded from 6 separate labeled regions and the average is calculated. The intensity after 30 second exposure is expressed as a percentage of the intensity at zero time. The values for PG are taken from the manufacturer s published results.
| Product | Counterstain | Catalog Number | Unit Size | Hardening | Refractive Index |
| VECTASHIELD Antifade Mounting Medium | None | H-1000 | 10 ml | No | 1.45 |
| DAPI | H-1200 | 10 ml | No | 1.45 | |
| PI | H-1300 | 10 ml | No | 1.45 | |
| VECTASHIELD PLUS Antifade Mounting Medium | None | H-1900 | 2 ml, 10 ml | No | 1.45 |
| DAPI | H-2000 | 2 ml, 10 ml | No | 1.45 | |
| VECTASHIELD HardSet Antifade Mounting Medium | None | H-1400 | 10 ml | Yes | 1.36 (initial) 1.46 (cured) |
| DAPI | H-1500 | 10 ml | Yes | 1.36 (initial) 1.46 (cured) | |
| TRITC-Phalloidin | H-1600 | 10 ml | Yes | 1.36 (initial) 1.46 (cured) | |
| VECTASHIELD Vibrance Antifade Mounting Medium | None | H-1700 | 2 ml, 10 ml | Yes | 1.38 (initial) 1.47 (cured) |
| DAPI | H-1800 | 2 ml, 10 ml | Yes | 1.38 (initial) 1.47 (cured) |
Check out this video on coverslipping tips for immunofluorescence slide mounting
Correctly coverslipping your slides ensures that your tissue specimens are preserved for optimal imaging and viewing. In this blog post, we will walk you through how to properly coverslip your slides to protect your samples.
As far as antifade mounting media go, one-size-fits-all solutions are few and far between. Here, we break down key factors you should consider when choosing the best antifade mounting medium for your experiment.
You’ve got lots of options when it comes to choosing a mounting media. Let us help you decide whether setting or non-setting mounting media will work best for your workflow in this Tips & Tricks article.
Applicable patents and legal notices are available at legal notices.
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