ProLong Glass Antifade Mounting Medium is a glycerin-based, rigid, ready-to-use mounting medium that can be For fluorescent labeling of cells or tissue samples on microscope slides or coverslips. This reagent does not contain DNA stain or the blue DNA stain NucBlue (Hoechst 33342) or the deep red DNA stain SYTOX Deep Red. ProLong glass mounting media has a refractive index of 1.52 after curing, similar to glass coverslips, and is compatible with immersion oil and oil immersion light microscopy, allowing for superior resolution and sensitivity. ProLong Glass Mountants are also designed to provide unparalleled photobleaching protection within the visible and near-infrared spectrum. For bright, high-resolution Z-Stack, 3D, and 2D images with image depths ranging from 0 to 100 µm, you can combine ProLong Glass Antifade Mounting Media with virtually any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry) and any cell/tissue type. For a non-curing mounting medium with a refractive index of 1.52, try SlowFade Glass Soft Antifade Mounting Medium.

ProLong Glass Anti-fade Mounting Media has a high refractive index of 1.52 and an excellent anti-fade formula, providing the following benefits:
•Up to 75% increased axial resolution for clear images
• Imageable focus depth 3–4 times that of refractive index 1.47 mounting media, enabling clear images of monolayers, tissue sections, or 3D cell cultures up to 150 μm thickness/focus depth
• Presents low spectral range Background
• Protects signal from inhibited photobleaching
• Slide-mounted samples remain stable for weeks to months
• Easy to use
• Available with or without DAPI

Learn more about ProLong Antifade Mounting Media>

ProLong Glass Rigid Antifade Mounting Media has Mixed, ready-to-use in dropper bottles or 10 mL bottles with or without DNA stain. NucBlue (Hoechst 33342) is excited by ultraviolet light when bound to DNA. The excitation wavelength is 360 nm, the upper emission wavelength is 460 nm, and DAPI traditional filter is used for detection. SYTOX Crimson Stain is excited by red light when it binds to DNA. The excitation wavelength is 660 nm and the upper limit emission wavelength is 682 nm. It is detected using the Cy5/Crimson traditional filter. After removing excess water, simply add a drop of mounting medium to the sample, add a coverslip, cure, and image. ProLong Glass Mountant is a hard-curing mounting medium that minimizes optical refraction and spherical aberration in the optical path, allowing for longer-term sample storage and helping to protect your samples from light bleach. ProLong Glass Mounting Medium does not discolor or shrink during the curing process, so high-quality fluorescence images can be obtained weeks or even months after the slide has been mounted.

ProLong Glass Antifade Mounting Medium with NucBlue Stain contains an optimized concentration of Hoechst 33342 stain to provide excellent nuclear labeling for most cells and tissues. ProLong Glass Mounting Media with NucBlue Stain is not recommended for specimens thicker than 30 µm. Thicker samples require longer incubations with labeling reagents, so NucBlue Live ReadyProbes reagent is recommended Perform pre-incubation.

Brighter and clearer images
Study shows that by matching the refractive index of the mounting medium, the axial resolution of the microscopy system can be improved to maximize Light collection by objective lenses (1–3). Fouquet et al. found that axial resolution improved significantly when using confocal microscopy with high refractive index media (3). When ProLong Glass Antifade Mounting Medium with a refractive index of 1.52 was compared to a mounting medium with a refractive index of 1.47, axial resolution improved by 32% and 75% at 40 µm and 100 µm depth of focus, respectively ( See below). As the depth of focus within the sample increases, the signal intensity and sensitivity of the fluorophore improves. Lateral resolution and sensitivity are also improved by equal or greater margins. This makes ProLong Glass Antifade Mountant an excellent mounting medium for acquiring Z-scan, 3D and 2D images (0 to 100 µm depth) of cells and tissues (see image below).

Inhibit photobleaching in the visible and near-infrared spectrum
No matter which fluorescent dye or fluorescent protein you are using, ProLong Glass Antifade Mounting Media can help Provides superior photobleaching protection. It has proven to have superior performance across a range of fluorescent dyes including Alexa Fluor, FITC, TRTC, Texas Red, Cy3 and Cy5 dyes (see figure below). ProLong glass mounting medium does not quench fluorophores, allowing fluorescent dyes to emit brighter signals without background signal, helping you obtain extremely high-quality, high-resolution images ready for publication.

Signal Stability During Imaging and Long-Term Storage
ProLong Glass Antifade Mounting Medium is ideal for archiving samples containing fluorescent dyes or fluorescent proteins and maintaining signal during prolonged imaging. Ideal for intensities such as when scanning repeatedly under confocal laser scanning illumination, or imaging with super-resolution microscopy. Easy to use—simply remove excess water from stained samples and cure.

Having an optimal refractive index that matches sharp images
For modern high-resolution imaging applications, refractive index matching is critical. ProLong Glass Antifade Mounting Media cures to a refractive index of 1.52. Additionally, this curing feature eliminates the need for the use of dirty acrylics (e.g., nail polish) that can damage or destroy modern microscope objectives.

References
1: Cremer C, Masters BR. Microscope resolution enhancement technology. Eur Phys J. 2013 38(3):281-344.
2: Shimi T, Kittisopikul M, Tran J, Goldman AE, Adam SA, Zheng Y, Jaqaman K, Goldman RD. Nuclear visualization by super-resolution microscopy Structural organization of layers A, C, B1 and B2. Mol Biol Cell.2015 26(22):4075-86.
3: Fouquet C, Gilles JF, Heck N, Dos Santos M, Schwartzmann R, Cannaya V, Morel MP, Davidson RS, Trembleau A, Bolte S. Improved axial resolution of confocal microscopy using new high-refractive index mounting media. PLoS One.2015 10(3):e0121096.