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:
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
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
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)
Mount sections onto subbed or charged microscope slides and air dry (in the dark) until sections are moist but all excess liquid has evaporated
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.
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
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ICC
Add a drop of mounting medium (Around 5µl for a 10mm and 15µl for a 22mm coverslip) to a standard microscope slide.
Briefly rinse the coverslip in dH2O before placing face down into the drop of mounting medium being careful not to introduce bubbles.
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.