The VECTASHIELD HardSet?Antifade?Mounting Medium with TRITC-Phalloidin?prevents rapid photobleaching of fluorescent proteins and fluorescent dyes.?Fluorescent derivatives of phalloidin are used to stain actin filaments. TRITC (tetramethylrhodamine) is excited at 544 nm and emits at 572 nm, producing an orange-red fluorescence.?VECTASHIELD HardSet Mounting Media hardens after coverslipping. After approximately 15 minutes at room temperature, the coverslip will become immobilized, and optimal antifade ability and refractive index will be achieved.

Features:

• Ideal refractive index (1.46)
• Ready-to-use – from the refrigerator to the benchtop
• No warming necessary
• Can be stored without sealing for long term analysis
• Hardening formulation

VECTASHIELD?Mounting Media are compatible with a wide array of fluorochromes, enzymatic substrates, and fluorescent proteins. Please consult the compatibility table (in the "Documents" section) to determine if VECTASHIELD?will be compatible in your system.

All formulations of?VECTASHIELD Antifade?Mounting Media are available with or without the counterstain DAPI (4', 6-diamidino-2-phenylindole). ?The DAPI concentration can be modified by mixing with the corresponding VECTASHIELD?Mounting Medium?without DAPI. ?DAPI produces a blue fluorescence when bound to DNA with excitation at about 360 nm and emission at 460 nm.

R.J. Florijn, et. al. Cytometry, 19 (1995) 177-182.

The refractive index for VECTASHIELD?Mounting Medium is 1.45.

VECTASHIELD?Mounting Medium in Super Resolution Microscopy

The optimal medium for super-resolution imaging methods maximizes the lifetime and photoswitching characteristics of fluorophores. VECTASHIELD?Mounting Medium has been shown to be compatible with a number of fluorophores used in super resolution methods including STORM, STED, and 3D-SIM imaging, such as Cy5 or Alexa Fluor 647, resulting in greater convenience and reproducibility of the method. (Olivier, N. et al., 2013, Biomedical Optics Express, Vol. 4 No. 6, pp. 885-899; Glushonkov, O. et al., 2018, Scientific Reports, Vol.?8,?8749?(2018)?).

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 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.