VECTASHIELD Vibrance Antifade Mounting Media are completely new formulations of curing (hard setting) antifade mounting media for immunofluorescence applications. These newest additions to our VECTASHIELD portfolio were developed with customer feedback to improve on parameters such as ease of use and retention of specific staining intensity over time. VECTASHIELD Vibrance Antifade Mounting Media are new tools to help investigators see more and do more with each experiment.
Unit Size | 2 ml, 10 ml |
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Applications | Immunofluorescence, In situ hybridization, Cellular Imaging |
Recommended Storage | Refrigerate |
Mounting | Aqueous (Hardening) |
Antifade | Yes |
Counterstain | None |
Results can be viewed after one hour, but two hours is recommended for optimal signal intensity.
With adequate removal of excess buffer, the VECTASHIELD Vibrance will cure enough by one hour to hold the coverslip in place. Complete curing takes about 4 – 24 hours at room temperature.
After mounting with VECTASHIELD Vibrance, slides can be stored at room temperature for several months without media retraction, bubble formation, or loss of signal intensity. We have not seen any difference between room temperature or storage at 4 degrees C.
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, no sealing of the coverslip is requuired.
VECTASHIELD Vibrance has outstanding antifade properties, acoss the spectrum – even under far-red wavelengths – and it is compatible with most commercially available fluorophores.
Competitor workflows for immunofluorescence recommend that slides be viewed 24 hours after mounting. VECTASHIELD Vibrance Mounting Media, however, allow same-day viewing. Accelerate discovery without sacrificing signal intensity or retention.
Above image: Improved retention of fluorescence with VECTASHIELD Vibrance Mounting Medium. Serial sections of human colon tissue (FFPE) stained for cytokeratin (mouse primary antibody AE1/AE3) and detected with fluorescein conjugated horse anti-mouse IgG secondary antibody (FI-2000). One hour after mounting with either VECTASHIELD Vibrance (top row) or a competitor mounting medium (bottom row), the sections were imaged at the intervals indicated. Note retention of fluorescent signal for section mounted with VECTASHIELD Vibrance Mounting Medium.
Extended Archiving of Mounted Sections at Room Temperature
Archiving fluorescently stained sections at room temperature often leads to signal reduction and dessication of the specimen. Slides mounted with VECTASHIELD Vibrance Medium, however, can be stored for weeks at ambient temperature, in standard slide boxes, with no loss of specific signal intensity or specimen integrity. The stabilizing effect of VECTASHIELD Vibrance Mounting Medium provides greater flexibility for imaging and storing fluorescently stained samples.
A problem commonly observed in immunoflourescence staining is the formation of bubbles under the coverslip, or the shrinking (retraction) of the media over time. This phenomenon detracts from the overall quality of the sections and can obscure specific staining. The unique formulation of VECTASHIELD Vibrance Mounting Medium provides a uniform sheet of medium under the coverslip, with essentially no bubble formation or retraction—even after many weeks of storage at room temperature.
Serial sections of human colon tissue (FFPE) stained for cytokeratin (AE1/AE3, mouse primary) followed by Alexa Fluor™ 488 anti-mouse IgG secondary antibody. Sections were mounted with either VECTASHIELD Vibrance Mounting Medium (H-1700, left image) or competitor mounting medium (right image) stored at room temperature for two weeks, and then imaged.
Note the absence of bubbles and background in green spectrum in left image (VECTASHIELD Vibrance Mounting Medium) compared with right image, after two weeks at room temperature.
Check out the video below on how to use a hydrophobic barrier pen.
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