VECTASHIELD® Antifade Mounting Medium with DAPI

Description

Still using the original VECTASHIELD with DAPI?

Try our upgraded product VECTASHIELD^® PLUS Antifade Mounting Medium with DAPI

• Description

• Specifications

• Documents

• Product FAQs

• Citations

• Technical Information

Description

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.

Features:

• Ideal refractive index
• Ready to use
• No warming necessary
• Can be stored without sealing for long term analysis
• Easy to use

Specifications

Unit Size	10 ml
Applications	Immunofluorescence, In situ hybridization, Cellular Imaging
Mounting	Aqueous (Non-Hardening)
Antifade	Yes
Counterstain	DAPI (blue fluorescence)
Cellular Stain	Nucleus

Documents

• VECTASHIELD Fluorochrome Compatibility
• User Guide
• In Situ Hybridization Detection Protocol
• Safety Data Sheet
• Download CoA
• Datasheet

Product FAQs

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Can VECTASHIELD® be used for super resolution microscopy?

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.

Does VECTASHIELD contain glycerol?

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.

Do I need to dehydrate the tissue sections before applying VECTASHIELD?

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.

Do I need to seal the perimeter of the coverslip after I apply 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.

Citations

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Technical Information

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.

VECTASHIELD Mounting Medium Antifade Comparison

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)

Additional Properties of VECTASHIELD Antifade Mounting Medium

Check out this video on coverslipping tips for immunofluorescence slide mounting

Step by Step Protocol