Immunofluorescence imaging is a useful laboratory technique for visualizing tissue structure, protein localization, and many other features of tissues and cells. Choosing the best mounting media is paramount for the success of immunofluorescence microscopy experiments. In order for this microscopy to provide useful data, the fluorescence on the slides must be maintained.
Vector Laboratories is one of the leading providers of reagents for immunofluorescence imaging. They provide premium labeling and detection products at each step of your immunofluorescence workflow. Vector Laboratories is a market leader for mounting media that provide excellent signal retention for image acquisition and specimen archiving. Vector Laboratories first introduced an antifade mounting media in 1993.
Recently, Vector launched two new mounting media: VECTASHIELD Vibrance and VECTASHIELD PLUS. These products mark the second generation of the best in class antifade mounting media, VECTASHIELD. They improve upon many pain points that researchers using immunofluorescence microscopy (including myself!) have long faced:
Flexibility with counterstaining
VECTASHIELD Vibrance and VECTASHIELD PLUS Antifade Mounting Media are both available in formulations with or without DAPI. The DAPI formulations of both products provide excellent contrast, with no background haze. This allows for superior signal interpretation and imaging compared to competitors’ mounting media with DAPI. It also saves time by circumventing the separate counterstaining step from conventional immunofluorescent staining protocols.
Retention of fluorescence signal
Fluorophores are susceptible to photobleaching and fading from the imaging excitation light, handling, and during storage. VECTASHIELD Vibrance and VECTASHIELD PLUS are superior to competitors’ mounting media for preserving fluorescence across the spectrum. Just one hour after applying the coverslip, slides mounted with VECTASHIELD Vibrance or VECTASHIELD PLUS can be visualized under the microscope, saving a lot of time! In fact, VECTASHIELD Vibrance hardens at room temperature in as little as 20 minutes. Conventional immunofluorescence workflows recommend slides be viewed 24 hours after mounting.
At the one hour time point, slides that were mounted with VECTASHIELD Vibrance or VECTASHIELD PLUS retained their fluorescence signal after being imaged for up to 180s, while slides mounted with a competitor product began to lose their fluorescence after just 60s. For example, one hour after mounting the coverslip with VECTASHIELD PLUS, Cy3 and Cy5 signals are maintained at >90% after 180s imaging, and FITC signal is maintained at 80%.
It is typically recommended that slides be stored at 4°C for the fluorescence to be optimally retained; however, refrigerator space is often limited in the laboratory setting. If slides are mounted with VECTASHIELD Vibrance, the fluorescence signal does not fade nor does the media retract or form bubbles, even when slides are stored at room temperature. Therefore, VECTASHIELD Vibrance allows slides to be stored at room temperature without sacrificing the integrity of data that can be obtained from the slides.
Slides mounted with VECTASHIELD PLUS are recommended to be stored at 4°C due to its non-hardening formulation. No sealing of the coverslips are required for short term storage. However, sealing of coverslips is recommended with long term archiving for retention of maximum antifade media performance.
Using either of these two mounting media formulations from Vector Laboratories is sure to make immunofluorescence imaging more efficient and produce cleaner data versus using competitors’ products. And to add an additional level of flexibility at the practical and financial levels, both products are available in either 2mL or 10mL sizes.
A great source for these products as well as all immunofluorescence reagents from Vector Laboratories is Quartzy!
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Aliyah is a science communicator with a PhD in immunology, currently based in Boston. You can find her on Twitter @desabsurdites and on her blog at www.aliyahweinstein.com.