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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.

ProLong™ Glass Antifade Mountant with NucBlue™ Stain (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.

Prionics™ Evans Blue Counterstain

Counterstain for immunofluorescence staining with FITC labeled antibodies.

SPoT-Light Tissue Pretreatment Kit

For Enzyme and Heat Pretreatment of FFPE Tissue prior to CISH.

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.

CytoVista™ Tissue Clearing Enhancer (Invitrogen™)

CytoVista Tissue Clearing Enhancer helps clear fluorescently labeled tissue to enable sharp and bright 3D fluorescent imaging at any tissue depth. When used following CytoVista Tissue Clearing Reagent, this reagent can clear almost any tissue type with depths up to 10 mm. CytoVista Tissue Clearing Enhancer can also be used as an imaging solution with stable storage of tissues for weeks with minimal change in tissue morphology or brightness, or specificity of the fluorophores. As an imaging solution, it has refractive index of 1.53.

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.

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.

AP Enhancer Solution (Invitrogen™)

AP Enhancer Solution, 40 mL is an aqueous buffered solution and an individual component of the ViewRNA™ ISH Tissue Assay Kits, 2-Plex, 96 assays. It is an enhancer solution for alkaline phosphatase.

CytoVista™ Permeabilization Buffer (Invitrogen™)

CytoVista Permeabilization Buffer is used while performing immunohistochemistry (IHC) or immuno-labeling with tissue. This buffer is highly recommended for larger, more collagenous, and difficult-to-clear tissues (e.g., liver tissue, archived (FFPE) tissue). It helps reagents penetrate thick tissue, helping enable sharp and bright images at any tissue depth 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.

CytoVista™ Tissue Clearing Kit (Invitrogen™)

The CytoVista Tissue Clearing Kit clears fluorescently labeled tissue to enable sharp and bright 3D fluorescent imaging at any tissue depth. The kit contains both CytoVista Tissue Clearing Reagent and CytoVista Tissue Clearing Enhancer, which together can be used for to clear most fluorescently labeled tissue types. CytoVista Tissue Clearing Reagent or Enhancer can also be used as an imaging solution with stable storage of tissues for weeks with minimal change in tissue morphology or brightness, or specificity of fluorophores.

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.

Image-iT™ FX Signal Enhancer ReadyProbes™ Reagent (Invitrogen™)

Image-iT® FX Signal Enhancer ReadyProbes® Reagent dramatically reduces background fluorescence resulting from non-specific staining during secondary detection using conjugates of streptavidin or secondary antibodies. This reagent is applied directly to slides or coverslips containing fixed and permeabilized cell or tissue samples prior to staining with fluorescent probes.

• Easy protocol—apply ready-made solution using a dropper bottle
• Compatible with the other steps in your immunostaining protocol
• Effective with most fluorescent labels
• Stable at room temperature—keep handy at your scope or cell culture area

See other ReadyProbes® reagents for cell staining
Find more tools for fixed cell imaging

Cell imaging applications
Image-iT® FX Signal Enhancer reagent is a highly effective product for blocking background staining that results from the nonspecific interactions of a wide variety of fluorescent dyes with cell and tissue constituents. Background staining seen with fluorescent conjugates of streptavidin and antibody conjugates are largely eliminated when the Image-iT® FX Signal Enhancer reagent is applied to fixed and permeabilized cells prior to staining.

Suggestions for use
• Apply Image-iT® FX Signal Enhancer reagent directly to slides or coverslips containing cell or tissue samples prior to immune-staining.
• After the fix/perm step, simply wash and add enough drops to cover your sample, then incubate for 30 min and proceed with your normal staining protocol.
• Additional blocking steps may be performed subsequent to blocking with the Image-iT™ Signal Enhancer ReadyProbes® Reagent, as needed

Image-iT™ Green Hypoxia Reagent (Invitrogen™)

Image-iT Green Hypoxia Reagent is a novel, fixable fluorogenic compound for measuring hypoxia in live cells. It is non-fluorescent when live cells are in an environment with normal oxygen concentrations and becomes fluorescent when oxygen levels are decreased. Image-iT Green Hypoxia Reagent sustains its fluorescence when cells/tissue return to normal oxygen levels, allowing the cells/tissue to be fixed with minimal loss of fluorescent signal.

Features of Image-iT Green Hypoxia Reagent include:
• Indicates hypoxia in live cells by fluorescing in low oxygen environments
• Fixable—sustains fluorescent signal when cells/tissue return to normal oxygen levels
• Easy to use—just add to cell culture media and image

Image-iT Green Hypoxia Reagent is a fluorogenic compound that is live cell permeable and becomes fluorescent in environments with low oxygen concentrations. These properties makes it a highly useful tool for detecting cells and tissue under hypoxic conditions. Image-iT Green Hypoxia Reagent is a very sensitive oxygen detector. Unlike pimonidazole adducts that respond only to very low oxygen levels, Image-iT Green Hypoxia Reagent begins to fluoresce when atmospheric oxygen levels are less than 5%. It responds quickly to such environments, and the fluorescence is sustained after the oxygen levels return to normal. These properties make Image-iT Green Hypoxia Reagent an ideal tool for detecting hypoxic conditions around tumors, 3D cultures, spheroids, neurons, etc.

ProLong™ Gold Antifade Mountant (Invitrogen™)

ProLong® 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, and delivers this protection without significantly quenching the initial fluorescence signal. ProLong® Gold Antifade Mountant is a curing mountant that forms a near-perfect optical path (1.47 RI) and allows longer-term storage of the sample. It comes ready-to-use (room temperatures storage and no mixing required)—just apply a drop to the sample, add a coverslip, cure, and image. ProLong® Gold mountant is available with or without DAPI nuclear stain.

ProLong® 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, ProLong® Diamond Antifade Mountant is recommended. Learn more about key differences between ProLong® Gold and ProLong® Diamond antifade mountants.

Key Attributes:

• Protects dyes from fading during imaging
• Ready-to-use liquid that cures for longer-term storage
• Mounted samples are stable for months
• Maintains fluorescence signal—little to no quenching
• Ideal for Alexa Fluor® dyes

Get Photobleaching Protection Without Quenching
Many mountants can destroy initial fluorescence signal strength 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.

Stable Signals During Imaging and for Longer-Term Storage
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. ProLong® Gold Antifade Mountant is the next-best option for archiving samples. And it is the best option for preserving signal strength during prolonged imaging sessions, such as repeated scanning under confocal laser scanning illumination. It's easy to use—just air-dry your stained samples to remove excess water and mount. For immediate viewing of the sample, choose our non-curing mountant, SlowFade® Gold Antifade Mountant.

Achieve Excellent Refractive Index Matching Without Acrylic Resins
For modern high-resolution imaging applications, refractive index matching is critical. ProLong® Gold Antifade Mountant will cure to an Rf of 1.47. Additionally, the curing property means you can avoid messy acrylic resins (i.e., finger nail polish), which can damage or ruin modern microscope objectives if it comes into contact with them.

Probe Set Diluent QT (Invitrogen™)

Probe Set Diluent QT, 12 mL is an aqueous solution containing formamide, detergent, and blocker. It is an individual component of the ViewRNA™ ISH Tissue Assay Kits, 2-Plex, 24 assays.

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.