MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset)

(HB8459)
Technical documents: Datasheet

Product overview

Name MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset)
Biological description

Overview

MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset) is an ideal formulation for prevention of photobleaching of fluorescent proteins and dyes during fluorescent imaging. It is easy to use with an ideal refractive index and provides effective prevention of photobleaching. This formulation contains DAPI which is a blue fluorescent DNA stain used to label cell nuclei.

Applications: IHC(IF), ICC, Cellular imaging, Super-resolution microscopy
Mounting: Aqueous (hardset) - cures in approximately 1 hour at room temperature
Antifade: Yes
Counterstain: DAPI
Refractive index: ≈1.45 (initial) which then increases to ≈1.518 once cured


Other Mounting Media Products

We supply a full range of mounting media for a range of experimental needs:

Hardset:

Aqueous:

Description

Antifade fluorescence hard-set mounting medium with DAPI for use in IHC(IF) and ICC.

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Images

Figure 1. Calbindin and GFAP staining in the cerebellum. Mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset)

4% PFA fixed 40µm horizontal rat brain sections were stained for Calbindin (HB6396) and GFAP(HB8001). Sections were mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset). For a full protocol and more information on IHC(IF) please see our IHC(IF) protocol

Figure 2. Calbindin and GFAP staining in the cerebellum. Mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset).

4% PFA fixed 40µm horizontal rat brain sections were stained for Calbindin (HB6396) and GFAP(HB8001). Sections were mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset). For a full protocol and more information on IHC(IF) please see our IHC(IF) protocol

Figure 3. Antifade performance of MightyMountTM Antifade Fluorescence Mounting Medium (hardset) compared to VECTASHIELD® and Fluoromount-G™

Hello Bio MightyMountTM Antifade Fluorescence Mounting Medium (hardset) shows strong antifade characteristics against a range of fluorophores and compares favorably against VECTASHIELD® and Fluoromount-G™. Method: HEK293T cells were cultured on coverslips then stained with either DAPI (HB0747), FITC Phalloidin (HB0814) or a β-tubulin (HB6491) with DyLight 488 or Janelia Fluor 549 conjugated secondary antibodies. Following mounting with either MightyMountTM Antifade Fluorescence Mounting Medium (hardset), VECTASHIELD® or Fluoromount-G™ slides were imaged on a confocal microscope. Imaging was carried out as a timeseries at 100% laser power with the normalized intensity being calculated compared to the first exposure.

Figure 4. Calbindin and GFAP staining in the cerebellum. Mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset)

4% PFA fixed 40µm horizontal rat brain sections were stained for Calbindin (HB6396) and GFAP(HB8001). Sections were mounted using MightyMountTM Antifade Fluorescence Mounting Medium with DAPI (hardset). For a full protocol and more information on IHC(IF) please see our IHC(IF) protocol

Optical Data

Fluorescence spectra Fluorescence Spectra
Emission color Blue
Max excitation wavelength 350nm
Max emission wavelength 470nm
Closest laser lines 405nm
Cell permeable Yes

Biological Data

Application notes Protocol for use of mounting media

Once mounted, store slides at 4°C in the dark for optimal preservation of fluorescence.

IHC(IF)
  1. Mount sections onto subbed or charged microscope slides and air dry (in the dark) until sections are moist but all excess liquid has evaporated
  2. Add a few drops of mounting media around the sections (around 50µl but this will depend on the number and thickness of sections) and slowly lower the coverslip from one end of the slide to the other being careful to avoid creating any bubbles.
  3. Wrap slides in foil to prevent light exposure then allow the media to cure at 4°C overnight before imaging. If more rapid imaging is needed it is possible to accelerate the curing process by incubating slides at either room temperature or 37°C for ≈1 hour.

For more information on IHC(IF) including tips on how to mount sections, please see our IHC(IF) protocol

 

ICC
  1. Add a drop of mounting medium (Around 5µl for a 10mm and 15µl for a 22mm coverslip) to a standard microscope slide.
  2. Briefly rinse the coverslip in dH2O before placing face down into the drop of mounting medium being careful not to introduce bubbles.
  3. Wrap slides in foil to prevent light exposure then allow the media to cure at 4°C overnight before imaging. If more rapid imaging is needed it is possible to accelerate the curing process by incubating slides at either room temperature or 37°C for ≈1 hour.

For more information on ICC please see our ICC protocol

Solubility & Handling

Storage instructions

+4°C or -20°C long-term. Protect from light.

Storage buffer

Contains 0.05% sodium azide

Important This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.

Calculators

Molarity

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Dilution

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FAQs

If I accidentally left the bottle out and the mounting media set what do I do?

If the mounting media has set prematurely then simply heat the bottle to around 60°C and the mounting media will re-liquify.