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SlowFade™ Diamond Antifade Mountant with DAPI (Invitrogen™)

SlowFade® Diamond Antifade Mountant is a glycerol-based liquid mountant applied directly to fluorescently labeled cells or tissue samples on microscope slides. SlowFade® Diamond mountant provides superior antifade protection across the entire visual and IR spectrums. You can use SlowFade® Diamond mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry)to achieve the brightest signal and lowest background.

Features of SlowFade® Diamond Antifade Mountant include:
• Protects fluorescent dyes and fluorescent protein from fading, across entire visible and IR spectrums
• Ready-to-use liquid that does not cure; samples can be viewed immediately
• Ideal for Alexa Fluor® and traditional dyes, such as FITC and Cy®3, and fluorescent proteins like GFP, RFP, and mCherry
• Maintains fluorescence signal—little to no quenching

Learn more about SlowFadfe® Antifade mountants.

SlowFade® Diamond Antifade Mountant comes already mixed and ready to use in a dropper bottle, with or without DAPI. Just apply a drop to the sample, add a coverslip, and image. SlowFade® Diamond mountant is a non-curing mountant that forms a near-perfect optical path (1.42 RI). When the coverslip edges are sealed with an appropriate coverslip sealant, samples can be stored for days to weeks. If long term storage is desired with a refractive index (RI) closer to immersion oil, then ProLong® Diamond Antifade Mountant is recommended. With the no-curing-required properties of SlowFade® Diamond mountant, samples can be evaluated and imaged right after labeling.

Get Photobleaching Protection Across the Entire Visible and IR Spectrums
No matter which fluorescent dyes or fluorescent proteins you are using, SlowFade® Diamond mountant helps provide the best protection against photobleaching. It is proven to have best-in-class performance against a wide array of fluorescent dyes, including Alexa Fluor® dyes, FITC, TRTC, Texas Red®, Cy®3, and Cy®5. SlowFade® Diamond does not quench, and its ability to provide brighter signal, against no background, helps ensure that you obtain the best-quality, high resolution images for publishing.

Stable Signals During Imaging and for Immediate Viewing
SlowFade® Diamond mountant is the ideal option for analyzing samples right after labeling. Its antifade properties for both fluorescent dyes and fluorescent proteins preserves signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination, or with super resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. A slide sealant can be used to preserve the slides for days or weeks. If longer archiving is desired, then the SlowFade® Diamond mountant can be washed away with PBS, and the samples remounted with ProLong® Diamond Antifade Mountant.

Achieve Excellent Refractive Index for Crisp Images
For modern high-resolution imaging applications, refractive index (RI) matching is critical. SlowFade® Diamond mountant has an RI of 1.42 , which is much closer to the RI of cells or tissue. For thick samples, it might be essential to use SlowFade® mountaint in order to obtain high resolution images.

ProLong™ Diamond Antifade Mountant with DAPI (Invitrogen™)

ProLong® Diamond Antifade Mountant is a glycerol-based liquid mountant applied directly to fluorescently labeled cells or tissue samples on microscope slides. ProLong® Diamond mountant is designed to provide unparalleled antifade protection across the entire visual and IR spectrums. You can use ProLong® Diamond mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry) to achieve the brightest signal and lowest background, for bright, high definition fluorescent images.

Features of ProLong® Diamond Antifade Mountant include:
• Protects fluorescent dyes and fluorescent protein from fading, across entire visible and IR spectrums
• Ready-to-use liquid that cures for longer-term storage
• Ideal for Alexa Fluor® and traditional dyes, such as FITC and Cy®3, and fluorescent proteins like GFP, RFP, and mCherry• Mounted samples are stable for months
• Maintains fluorescence signal—little to no quenching

Learn more about ProLong® Antifade mountants.

ProLong® Diamond Antifade Mountant comes already mixed and ready to use in a dropper bottle, with or without DAPI. Just apply a drop to the sample, add a coverslip, cure, and image. ProLong® Diamond mountant is a curing mountant that forms a near-perfect optical path (1.47 RI) and enables longer-term storage of your samples, while protecting them from photobleaching. ProLong® Diamond mountant does not discolor or shrink when cured and stored long term, making it possible to take high quality fluorescent images weeks or even months after mounting the slides.

Get Photobleaching Protection Across the Entire Visible and IR Spectrums
No matter which fluorescent dyes or fluorescent proteins you are using, ProLong® Diamond mountant helps provide superior protection against photobleaching. It is proven to have best-in-class performance with a wide array of fluorescent dyes, including Alexa Fluor® dyes, FITC, TRTC, Texas Red®, Cy®3, and Cy®5. ProLong® Diamond mountant does not quench fluorophores, and its ability to provide brighter signal, against no background, helps ensure that you get the best-quality, high resolution images for publishing.

Stable Signals During Imaging and for Longer-term Storage
ProLong® Diamond mountant is the ideal option for archiving samples with fluorescent dyes or fluorescent proteins. It is also an ideal option for preserving signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination, or with super resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. For immediate viewing of the sample, choose our non-curing mountant, SlowFade® Diamond Antifade Mountant.

Achieve Excellent Refractive Index Matching for Crisp Images
For modern high-resolution imaging applications, refractive index (RI) matching is critical. ProLong® Diamond mountant will cure to an RI of 1.47. Additionally, this curing property means you do not need to use messy acrylic resins (i.e., finger nail polish), which can damage or ruin modern microscope objectives if it comes into contact with them.

ProLong™ Live Antifade Reagent, for live cell imaging (Invitrogen™)

ProLong® Live Antifade Reagent helps prevent the loss of fluorescent signal due to photobleaching during live cell imaging. When dyes are illuminated, they can degrade (usually permanently), reducing the sample’s fluorescent intensity and leading to the creation of free radical singlet oxygen, an element that can further degrade neighboring dye molecules. ProLong® Live Antifade Reagent metabolizes this element, increasing the signal intensity and duration while keeping the background signal low. ProLong® Live reagent has little-to-no effect on cellular viability, proliferation, or other cellular functions in experiments up to 48 hours.

Features of ProLong® Live Antifade Reagent include:
• Photobleach protection in live cell experiments, for fluorophores such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker®, and CellTracker®dyes
• Little-to-no effect on cellular viability, proliferation, or other cellular functions
• Easy to use, just add to the imaging solution or complete cell culture media or buffer, incubate, and image

ProLong® Live Antifade Reagent contains enzymes from the plasma membrane of naturally occurring E. coli. These enzymes metabolize the elements that can cause photobleaching. They work in the extracellular space and are not known to enter cells, so intracellular functions are minimally affected by ProLong® Live Antifade Reagent. As shown in figure below, cells grown in the presence of ProLong® Live Antifade Reagent display minimal effects on cellular viability or proliferation. ProLong® Live Antifade Reagent provides photobleach protection for most fluorophores frequently used in live cell fluorescent imaging, such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker® and CellTracker® dyes. ProLong® Live Antifade Reagent can be added to any cell culture media or buffer that is suitable for fluorescent imaging.

SlowFade™ Gold Antifade Mountant (Invitrogen™)

SlowFade® Gold Antifade Mountant is a liquid mountant applied directly to fluorescently labeled cell or tissue samples on microscope slides. It contains chemical components designed to protect fluorescent dyes from fading (photobleaching) during fluorescence microscopy experiments. SlowFade® Gold Antifade Mountant is glycerol-based, with no mixing or curing required, which makes it ideal for immediate viewing of the sample. It comes ready-to-use—just apply a drop to the sample, add a coverslip, and image. It is available with or without DAPI nuclear stain.

SlowFade® Gold Antifade Mountant is not recommended for mounting samples containing fluorescent proteins, such as GFP. For superior antifade protection of fluorescent proteins and fluorescent dyes, SlowFade® Diamond Antifade Mountant is recommended. Learn more about key differences between SlowFade® Gold and SlowFade® Diamond antifade mountants.

Key Attributes:

• Protects dyes from fading during imaging
• Ready-to-use liquid, ideal for immediate sample viewing
• Mounted samples are stable for weeks
• Maintains signal strength—little to no quenching
• Ideal for Alexa Fluor® dyes

Get Photobleaching Protection Without Quenching

The antifade reagents used in some mountants can destroy the initial strength of many fluorophores (especially those that emit above 550 nm) by up to 80%. To create the Molecular Probes® Gold series of mountants, our scientists developed a formula of chemical additives that minimize this initial signal quenching for the broadest range of colors and dyes.

Immediate Viewing with Maximum Signal Protection
While epoxy-based mountants used in our FluoCells® series of prepared cells can be stable for years and afford the highest level of antifade protection, they are difficult to use, requiring a series of steps in organic solvents prior to mounting. SlowFade® Gold Antifade Mountant is the best option for immediate viewing of samples. For longer-term storage, it is recommended that a sealant (i.e., finger nail polish or equivalent acrylic reagent) be applied to the coverslip edges to prevent drying. Alternatively, choose our curing mountant ProLong® Gold Antifade Mountant.

Improved Resolution in Thick Sections
Glycerol-based mountants are recommended when viewing thick samples, as the refractive index of glycerol is closer to that of tissue. It's easy to use—just air-dry your stained samples to remove excess water and mount.

CytoVista™ 10X Wash Buffer (Invitrogen™)

CytoVista 10X Wash Buffer is used in wash steps when performing immunohistochemistry (IHC) or immuno-labeling with tissue or 3D cell culture. This buffer is 10X concentrated and should be diluted to 1X with PBS before use. It consists of PBS with 0.2% Tween-20 and 10 µg/mL heparin and helps enable sharp and bright images at any depth of tissue or 3D cell culture when used with other CytoVista products.

Learn more about CytoVista 3D Culture Clearing reagents ›

Features of the CytoVista family of tissue and 3D cell culture clearing reagents include:
• Rapid clearing of fluorescently labeled tissue for 3D imaging
• Minimal shrinkage, expansion, or other morphological changes to tissue
• Simple workflow does not require any special instruments or equipment and is easily adapted to IHC
• Compatible with most fluorophores, including dyes and fluorescent proteins

The CytoVista tissue and 3D cell culture clearing workflow clears tissue or 3D cultured cells with minimal changes to morphology or the sensitivity of detection with almost any fluorophore. With an easy-to-use protocol, whole mouse brain (8-mm section) can be cleared in 24 hours, or a 1-mm section in 2 hours, without the use of any special instruments or equipment.

The CytoVista clearing workflow is compatible with most fluorophores, including fluorescent proteins that are detected with common fluorescent imaging instruments, such as wide-field microscopes, high content analyzers, confocal microscopes, and lightsheet microscopes. For first time users, we recommend the CytoVista Tissue Clearing/Staining Kit (Cat No V11324) or CytoVista 3D Cell Culture Clearing/Staining Kit (Cat No. V11325). These kits contain all necessary reagents for both clearing and IHC staining.

CytoVista clearing technology sufficiently clears tissue for 3D fluorescent imaging, but unlike other clearing techniques does not significantly changes the tissue morphology. Minimal morphological changes, such as shrinkage and contraction, have been observed. For precious samples, the clearing can be reversed, and the tissue can be processed for histology studies, like H&E staining.

For tissues that are easier to clear, like brain, lung, intestine, skin, and tissues of less than 250-µm thickness, treatment with CytoVista Tissue Clearing Reagent is sufficient for 3D imaging. However, tissues of more than 250-µm thickness or that are hard to clear like kidney, liver, heart, and placenta will require an initial treatment with CytoVista Tissue Clearing Reagent followed by treatment with CytoVista Tissue Clearing Enhancer.

CytoVista 3D Cell Culture Clearing Reagent is recommended for spheroids or organoids up to 1 mm in thickness.

For hard tissues like bone, cartilage, and connective tissue, CytoVista Tissue Clearing Reagent is not recommended.

Endogenous Biotin-Blocking Kit (Invitrogen™)

The Endogenous Biotin-Blocking Kit provides a method and reagents for a pretreatment that reduces or eliminates background signals when biotin-avidin or biotin-streptavidin detection systems are used to identify cellular targets.

The Endogenous Biotin-Blocking Kit offers:
• Improved results—increased signal-to-noise ratio facilitates cell target identification
• Ease of use—pipette-free dropper bottle offers speed and convenience
• Fast—endogenous background reduction in under an hour
• Flexibility—the kit can be used to help improve results from conventional immunohistochemistry, immunocytochemistry, RNA FISH, and ELISA procedures

Biotin occurs naturally, functioning as an enzyme cofactor in the cytosol and mitochondria of a wide variety of cell types. The Endogenous Biotin-Blocking Kit minimizes interference from the endogenous biotin found in enzymes of mammalian cells and tissues, through a two-step blocking strategy. First, an excess of unlabeled streptavidin is added to the specimen to bind endogenous biotin-rich enzymes. The streptavidin is subsequently blocked with an excess of unlabeled biotin, effectively rendering the cell or tissue sample free of available biotin-binding sites.

CytoVista™ 2-mm Sagittal Mouse Brain Slicer (Invitrogen™)

This 2-mm sagittal mouse brain slicer allows you to easily cut a mouse brain into smaller pieces for easier imaging with objectives that have a limited working distance. This slicer is 3D-printed out of resin and can be easily washed and cleaned between uses. All you need is a standard razor blade to start slicing your tissues.

Learn more about CytoVista 3D Culture Clearing reagents ›

Features of the CytoVista family of tissue and 3D cell culture clearing reagents include:
• Rapid clearing of fluorescently labeled tissue for 3D imaging
• Minimal shrinkage, expansion, or other morphological changes to tissue
• Simple workflow does not require any special instruments or equipment and is easily adapted to IHC
• Compatible with most fluorophores, including dyes and fluorescent proteins

The CytoVista tissue and 3D cell culture clearing workflow clears tissue or 3D cultured cells with minimal changes to morphology or the sensitivity of detection with almost any fluorophore. With an easy-to-use protocol, whole mouse brain (8-mm section) can be cleared in 24 hours, or a 1-mm section in 2 hours, without the use of any special instruments or equipment.

The CytoVista clearing workflow is compatible with most fluorophores, including fluorescent proteins that are detected with common fluorescent imaging instruments, such as wide-field microscopes, high content analyzers, confocal microscopes, and lightsheet microscopes. For first time users, we recommend the CytoVista Tissue Clearing/Staining Kit (Cat No V11324) or CytoVista 3D Cell Culture Clearing/Staining Kit (Cat No. V11325). These kits contain all necessary reagents for both clearing and IHC staining.

CytoVista clearing technology sufficiently clears tissue for 3D fluorescent imaging, but unlike other clearing techniques does not significantly changes the tissue morphology. Minimal morphological changes, such as shrinkage and contraction, have been observed. For precious samples, the clearing can be reversed, and the tissue can be processed for histology studies, like H&E staining.

For tissues that are easier to clear, like brain, lung, intestine, skin, and tissues of less than 250-µm thickness, treatment with CytoVista Tissue Clearing Reagent is sufficient for 3D imaging. However, tissues of more than 250-µm thickness or that are hard to clear like kidney, liver, heart, and placenta will require an initial treatment with CytoVista Tissue Clearing Reagent followed by treatment with CytoVista Tissue Clearing Enhancer.

CytoVista 3D Cell Culture Clearing Reagent is recommended for spheroids or organoids up to 1 mm in thickness.

For hard tissues like bone, cartilage, and connective tissue, CytoVista Tissue Clearing Reagent is not recommended.

ProLong™ Diamond Antifade Mountant (Invitrogen™)

ProLong® Diamond Antifade Mountant is a glycerol-based liquid mountant applied directly to fluorescently labeled cells or tissue samples on microscope slides. ProLong® Diamond mountant is designed to provide unparalleled antifade protection across the entire visual and IR spectrums. You can use ProLong® Diamond mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry) to achieve the brightest signal and lowest background, for bright, high definition fluorescent images.

Features of ProLong® Diamond Antifade Mountant include:
• Protects fluorescent dyes and fluorescent protein from fading, across entire visible and IR spectrums
• Ready-to-use liquid that cures for longer-term storage
• Ideal for Alexa Fluor® and traditional dyes, such as FITC and Cy®3, and fluorescent proteins like GFP, RFP, and mCherry• Mounted samples are stable for months
• Maintains fluorescence signal—little to no quenching

Learn more about ProLong® Antifade mountants.

ProLong® Diamond Antifade Mountant comes already mixed and ready to use in a dropper bottle, with or without DAPI. Just apply a drop to the sample, add a coverslip, cure, and image. ProLong® Diamond mountant is a curing mountant that forms a near-perfect optical path (1.47 RI) and enables longer-term storage of your samples, while protecting them from photobleaching. ProLong® Diamond mountant does not discolor or shrink when cured and stored long term, making it possible to take high quality fluorescent images weeks or even months after mounting the slides.

Get Photobleaching Protection Across the Entire Visible and IR Spectrums
No matter which fluorescent dyes or fluorescent proteins you are using, ProLong® Diamond mountant helps provide superior protection against photobleaching. It is proven to have best-in-class performance with a wide array of fluorescent dyes, including Alexa Fluor® dyes, FITC, TRTC, Texas Red®, Cy®3, and Cy®5. ProLong® Diamond mountant does not quench fluorophores, and its ability to provide brighter signal, against no background, helps ensure that you get the best-quality, high resolution images for publishing.

Stable Signals During Imaging and for Longer-term Storage
ProLong® Diamond mountant is the ideal option for archiving samples with fluorescent dyes or fluorescent proteins. It is also an ideal option for preserving signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination, or with super resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. For immediate viewing of the sample, choose our non-curing mountant, SlowFade® Diamond Antifade Mountant.

Achieve Excellent Refractive Index Matching for Crisp Images
For modern high-resolution imaging applications, refractive index (RI) matching is critical. ProLong® Diamond mountant will cure to an RI of 1.47. Additionally, this curing property means you do not need to use messy acrylic resins (i.e., finger nail polish), which can damage or ruin modern microscope objectives if it comes into contact with them.

ProLong™ Live Antifade Reagent, for live cell imaging (Invitrogen™)

ProLong® Live Antifade Reagent helps prevent the loss of fluorescent signal due to photobleaching during live cell imaging. When dyes are illuminated, they can degrade (usually permanently), reducing the sample’s fluorescent intensity and leading to the creation of free radical singlet oxygen, an element that can further degrade neighboring dye molecules. ProLong® Live Antifade Reagent metabolizes this element, increasing the signal intensity and duration while keeping the background signal low. ProLong® Live reagent has little-to-no effect on cellular viability, proliferation, or other cellular functions in experiments up to 48 hours.

Features of ProLong® Live Antifade Reagent include:
• Photobleach protection in live cell experiments, for fluorophores such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker®, and CellTracker®dyes
• Little-to-no effect on cellular viability, proliferation, or other cellular functions
• Easy to use, just add to the imaging solution or complete cell culture media or buffer, incubate, and image

ProLong® Live Antifade Reagent contains enzymes from the plasma membrane of naturally occurring E. coli. These enzymes metabolize the elements that can cause photobleaching. They work in the extracellular space and are not known to enter cells, so intracellular functions are minimally affected by ProLong® Live Antifade Reagent. As shown in figure below, cells grown in the presence of ProLong® Live Antifade Reagent display minimal effects on cellular viability or proliferation. ProLong® Live Antifade Reagent provides photobleach protection for most fluorophores frequently used in live cell fluorescent imaging, such as GFP, RFP, Hoechst 33342, MitoTracker®, LysoTracker® and CellTracker® dyes. ProLong® Live Antifade Reagent can be added to any cell culture media or buffer that is suitable for fluorescent imaging.

Image-iT™ FX Signal Enhancer (Invitrogen™)

Image-iT® FX signal enhancer is a liquid that is applied directly to slides or coverslips containing fixed and permeabilized cell or tissue samples prior to staining with fluorescent probes. When Image-iT® FX signal enhancer is used, nonspecific fluorescence (background) commonly seen with the application of fluorescent conjugates of streptavidin, goat anti-mouse, or goat anti-rabbit IgG is largely eliminated.

Key Attributes

Easy protocol—apply enhancer from ready-made solution in dropper bottle
Effective with many fluorescent labels (see Table 2 in user manual below)
Cost effective—supplies enough for 50 standard coverslips
Compatible with your immunostaining protocol’s other blocking steps

Reduce Nonspecific Binding of Dyes to Tissues.
Alexa Fluor® dyes and many other dyes have negatively charged modifications that can lead to nonspecific association with cells and tissue. While not always significant, it can be a problem when pushing the limits of sensitivity in low-abundance targets. Image-iT® FX signal enhancer can significantly reduce this binding, resulting in marked improvements in staining.

Need help deciding what product you need for your cell imaging? See our cell imaging workflow and decision tree.

Tired of poor signal-to-noise? See our collection antifade and signal enhancer products from Molecular Probes®.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

SlowFade™ Diamond Antifade Mountant (Invitrogen™)

SlowFade® Diamond Antifade Mountant is a glycerol-based liquid mountant applied directly to fluorescently labeled cells or tissue samples on microscope slides. SlowFade® Diamond mountant provides superior antifade protection across the entire visual and IR spectrums. You can use SlowFade® Diamond mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry) to achieve the brightest signal and lowest background.

Features of SlowFade® Diamond Antifade Mountant include:
• Protects fluorescent dyes and fluorescent protein from fading, across entire visible and IR spectrums
• Ready-to-use liquid that does not cure; samples can be viewed immediately
• Ideal for Alexa Fluor® and traditional dyes, such as FITC and Cy®3, and fluorescent proteins like GFP, RFP, and mCherry
• Maintains fluorescence signal—little to no quenching

Learn more about SlowFadfe® Antifade mountants.

SlowFade® Diamond Antifade Mountant comes already mixed and ready to use in a dropper bottle, with or without DAPI. Just apply a drop to the sample, add a coverslip, and image. SlowFade® Diamond mountant is a non-curing mountant that forms a near-perfect optical path (1.42 RI). When the coverslip edges are sealed with an appropriate coverslip sealant, samples can be stored for days to weeks. If long term storage is desired with a refractive index (RI) closer to immersion oil, then ProLong® Diamond Antifade Mountant is recommended. With the no-curing-required properties of SlowFade® Diamond mountant, samples can be evaluated and imaged right after labeling.

Get Photobleaching Protection Across the Entire Visible and IR Spectrums
No matter which fluorescent dyes or fluorescent proteins you are using, SlowFade® Diamond mountant helps provide the best protection against photobleaching. It is proven to have best-in-class performance against a wide array of fluorescent dyes, including Alexa Fluor® dyes, FITC, TRTC, Texas Red®, Cy®3, and Cy®5. SlowFade® Diamond does not quench, and its ability to provide brighter signal, against no background, helps ensure that you obtain the best-quality, high resolution images for publishing.

Stable Signals During Imaging and for Immediate Viewing
SlowFade® Diamond mountant is the ideal option for analyzing samples right after labeling. Its antifade properties for both fluorescent dyes and fluorescent proteins preserves signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination, or with super resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. A slide sealant can be used to preserve the slides for days or weeks. If longer archiving is desired, then the SlowFade® Diamond mountant can be washed away with PBS, and the samples remounted with ProLong® Diamond Antifade Mountant.

Achieve Excellent Refractive Index for Crisp Images
For modern high-resolution imaging applications, refractive index (RI) matching is critical. SlowFade® Diamond mountant has an RI of 1.42 , which is much closer to the RI of cells or tissue. For thick samples, it might be essential to use SlowFade® mountaint in order to obtain high resolution images.

Citrate Buffer (pH 6.0), Concentrate

Citrate buffer is used in Heat Induced Epitope Retrieval (HIER) methods. HIER is used to reverse the loss of antigenicity that occurs with some epitopes in formalin-fixed paraffin embedded tissues. During epitope retrieval, tissue slides are immersed in a solution and heated (>95° C) by hot plate or other appropriate heat source such as a pressure cooker or steamer. Citrate buffer is the preferred solution for most antibodies.

ProLong™ Glass Antifade Mountant (Invitrogen™)

ProLong Glass Antifade Mountant is a glycerol-based, hard-setting, ready-to-use mountant that can be applied directly to fluorescently labeled cells or tissue samples on microscope slides or coverslips. Prolong Glass mountant has a refractive index of 1.52 after curing, similar to that of glass coverslips, compatible immersion oil, and oil-immersion microscope optics, enabling superior resolution and sensitivity. ProLong Glass mountant is also designed to provide unparalleled photobleach protection across the visible and near infra-red spectra. You can use ProLong Glass Antifade Mountant with almost any fluorescent dye or fluorescent protein (e.g., GFP, RFP, mCherry) and with any cell/tissue type for image depths ranging from 0 to 100 µm for bright, high-resolution Z-Stack, 3D, and 2D images.

With its high refractive index of 1.52 and superior antifade formulation, ProLong Glass Antifade Mountant delivers the following benefits:
• Up to 75% improvement in axial resolution for sharp images
• 3–4 times more imageable focal depth compared tomountants with refractive index of 1.47, with sharp images for monolayers of cells, tissue slices, or 3D cell cultures up to 150 μm thickness/focal depth
• Low background across the spectrum
• Preserved signals from inhibited photobleaching
• Weeks to months of stability for slide-mounted samples
• Ease of use
• Availability with or without DAPI

Learn more about ProLong Antifade mountants >

ProLong GlassAntifade Mountant comes already mixed and ready to use in a dropper bottle or 10-mL bottle with or without NucBlue (Hoechst 33342) stain, a cell-permeant nuclear counterstain that emits blue fluorescence when bound to DNA. After removing excess water, just apply a drop of mountant to the sample, add a coverslip, cure, and image. ProLong Glass mountant is a hard-setting, curing mountant that minimizes optical refraction and spherical aberration in the optical path, enables longer-term storage of your samples, and helps protect your samples from photobleaching. ProLong Glass mountant does not discolor or shrink when cured, making it possible to take high-quality fluorescent images weeks, or even months, after mounting the slides.

ProLong Glass Antifade Mountant with NucBlue stain contains an optimized concentration of Hoechst 33342 stain, which provides superior nuclear labeling of most cells and tissues. ProLong Glass mountant with NucBlue stain is not recommended for specimens thicker than 30 µm. Thicker specimens can require longer incubations with labeling reagents, so a pre-incubation with NucBlue Live ReadyProbes Reagent is recomended.

Brightest and sharpest images
Studies show that the axial resolution of a microscopy system can be improved by matching the refractive index of the mounting medium, which maximizes light collection of the objective lens (1–3). Fouquet et. al. saw dramatic improvement in axial resolution while using a confocal microscope with high refractive index media (3). When Prolong Glass Antifade Mountant, which has a refractive index of 1.52, is compared with a mounting medium with a refractive index of 1.47, the axial resolution is improved by 32% at a focal depth of 40 µm and 75% at a focal depth of 100 µm (see figure below). The signal intensity and sensitivity of fluorophores is also improved as the focal depth increases within the sample. The same or greater improvement in resolution and sensitivity is seen in the lateral direction. This makes Prolong Glass a superior mountant for acquisition of Z-Stack, 3D, and 2D images of cells and tissue at depths from 0 to 100 µm (see figure below).

Inhibit photobleaching across the visible and near-IR spectra
No matter which fluorescent dyes or fluorescent proteins you are using, ProLong Glass Antifade Mountant helps provide superior protection against photobleaching. It has been shown to have best-in-class performance with a wide array of fluorescent dyes, including Alexa Fluor, FITC, TRTC, Texas Red, Cy3, and Cy5 dyes (see figure below). ProLong Glass mountant does not quench fluorophores, and its ability to provide brighter signal against no background helps ensure that you get the best-quality high-resolution images for publishing.

Stable signals during imaging and for longer-term storage
ProLong Glass Antifade Mountant is the ideal option for archiving samples with fluorescent dyes or fluorescent proteins and for preserving signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination or with super-resolution microscopy. And it's easy to use—simply remove excess water from the stained sample and mount. For immediate viewing of the sample, choose our non-curing mountant: SlowFade Diamond Antifade Mountant.

Achieve excellent refractive index matching for crisp images
For modern high-resolution imaging applications, refractive index matching is critical. ProLong Glass Antifade Mountant cures to a refractive index of 1.52. Additionally, this curing property eliminates the need to use messy acrylic resins (i.e., fingernail polish), which can damage or ruin modern microscope objectives.

References
1: Cremer C, Masters BR. Resolution enhancement techniques in microscopy. 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. Structural organization of nuclear lamins A, C, B1, and B2 revealed by superresolution microscopy. 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. Improving axial resolution in confocal microscopy with new high refractive index mounting media. PLoS One. 2015 10(3):e0121096.

SlowFade™ Antifade Kit (Invitrogen™)

The SlowFade® Antifade Kit contains two formulations of the SlowFade® Antifade Reagent: an aqueous formulation in buffer and an aqueous formulation that contains 50% glycerol. These mountants are applied directly to fluorescently labeled cell or tissue samples on microscope slides to protect the dyes from fading (photobleaching) during fluorescence microscopy experiments. Both formulations reduce the photobleaching rate of fluorescein to almost zero and are especially useful for quantitative measurements.

Key Attributes

Glycerol-based mountant, ideal for immediate sample viewing

Contains oxygen scavenger 1,4-diaza bicyclo (2.2.2) octane (DABCO)

Improved Signal Stability for Fluorescein
Use of the SlowFade® Antifade Kit can extend the useful fluorescence emission of fluorescein more than 50-fold, and can preserve the signal in cell and tissue mounts for up to two years. However, this original SlowFade® reagent formulation, like all DABCO-based mountants substantially quenches the initial fluorescence of fluorescein and almost completely quenches that of the Alexa Fluor® 350, Alexa Fluor® 405, and Cascade Blue® fluorophores.

Other Options for Antifade Mountants
New formulations of curing mountants (ProLong® Gold reagent) and glycerol-based mountants (SlowFade® Gold reagent) are now available with and without DAPI nuclear stain. These deliver improved performance for most dyes, including Alexa Fluor® dyes.