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AF1
AF1 is a mountant solution composed of glycerol, phosphate buffered saline and an antifadent. It was specifically designed to stop the photobleaching of the fluorescein moiety of FITC labelled biological specimens. Its application is not however limited to FITC labelled materials and has been used with advantage with many other fluorochromes including rhodamines and DAPI. It is ideal for examining tissue sections and dead cells. The solution has a pH of ~10. The solution should be pipetted onto the specimen and then a cover slip applied. If the slides are stored in a refrigerator, the viscosity of the mountant solution increases thereby helping too keep the cover slip in place. There is no need to seal the cover slip with nail varnish. Specimens mounted in AF1 solutions have been kept in this way for many months without suffering damage. Obtaining the correct viscosity for your application If the viscosity of the AF1 solution is too high for your purposes, it may be admixed with AF3 mountant solution. As the amount of AF 3 solution is increased so the viscosity decreases. Conversely, if you wish to have a higher viscosity add AF2 mountant solution to the AF1 solution. Increasing the amount of AF2 solution increases the viscosity - see below. |
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If you think this trick may be of value, why not order 1X25ml AF1 together with 1 X 25ml AF3 and obtain a discount? Details on Orders pages. Application Sheet AF1 (PDF 63KB) |
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AF2
AF2 is a mountant solution composed of glycerol and an antifadent. It was specifically designed to stop the photobleaching of the fluorescein moiety of FITC labelled biological specimens. Its application is not however limited to FITC labelled materials and has been used with advantage with many other fluorochromes including rhodamines and DAPI. It is ideal for examining tissue sections and dead cells. An aqueous solution (75%AF2 to 25% water v/v) has a pH of ~10. The solution should be pipetted onto the specimen and then a cover slip applied. If the slides are stored in a refrigerator, the viscosity of the mountant solution increases thereby helping too keep the cover slip in place. There is no need to seal the cover slip with nail varnish. Specimens mounted in AF2 solutions have been kept in this way for many months without suffering damage. Obtaining the correct viscosity for your application If the viscosity of the AF2 solution is too high for your purposes, it may be admixed with either AF1 or AF3 mountant solution. As the amount of AF1 or AF3 solution is increased so the viscosity decreases.
Application Sheet AF2 (PDF 62KB) |
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AF3
AF3 is a mountant solution composed of phosphate buffered saline and an antifadent. It was specifically designed to stop the photobleaching of the fluorescein moiety of FITC labelled biological specimens. Its application is not however limited to FITC labelled materials and has been used with advantage with many other fluorochromes including rhodamines and DAPI. It is useful for examining tissue live cells although it should be noted that cell lysis will take place over a period of time. The solution has a pH of ~10. The solution should be pipetted onto the specimen and then a cover slip applied. Specimens mounted in AF3 solutions have been kept in this way for many months without suffering damage. Application Sheet AF3 (PDF 58KB) |
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AF4
AF4 is a mountant solution composed of glycerol and
the antifadent n-propyl gallate. N-Propyl
gallate is a well established antifadent and is particularly useful
for stopping the photobleaching of DAPI and Alexa dye, Cye (cyanine)
dyes, Hoechst dye stained as well as FITC labelled materials. It
is ideal for examining tissue sections and dead cells. An aqueous
solution (75%AF4 to 25% water v/v) has a pH of ~5. AF4 may be
mixed with buffer solutions in order to attain the correct pH for the
fluorochrome being used but the resulting solutions may start to show
discolouration within a few hours due to oxidation of the phenol. Application Sheet AF4 (PDF 30KB) |
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AF87
AF 87 is a non-fluorescent immersion
oil of medium viscosity and has a refractive index of 1.516, and may
be used as an immersion oil or as a mountant medium. Application Sheet AF87 (PDF 30KB) |
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AF100 + CFPVOH
AF100 is a solution of an antifadent in phosphate buffered saline and CFPVOH is an aqueous solution of poly(vinyl alcohol). Addition of AF100 to CFPVOH gives a solution which on drying yields a clear semi-hard film which serves as a permanent mountant. The presence of the antifadent helps to reduce photobleaching of the fluorophore. Solutions should be made up by mixing 1 part by volume of AF100 with 9 parts by volume ofCFPVOH. These solutions should be used within 10 hours as the efficacy of the antifadent reduces with time. A few drops of the solution should be applied to the specimen followed by a cover slip. The water evaporates to give a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet AF100 + CFPVOH (PDF 58KB) |
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AF200 + Permafix-1 or UVM-2
AF200 is a solution of an antifadent in glycerol and has been specially prepared for Permafix PF and UVM-2 solutions where DAPI is the staining material. The solutions may be used with other fluorochromes such as fluorescein, Alexa dyes and Hoechst dyes. Suitable for Cye (cyanine) dyes. The presence of the antifadent helps to reduce photobleaching of the fluorophore. Solutions should be made up by mixing 1 part by volume of AF200 with 3 parts by volume of Permafix-1 or UVM-2. The solution made with Permafix-1 should be used within 10 hours as the efficacy of the antifadent reduces with time. A few drops of the solution should be applied to the specimen followed by a cover slip. When used with Permafix PF evaporation of the water leaves a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. The mixture of AF200 and UVM-2 may be cured with light in the usual way to give a clear film. |
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AF300 + UVM-2
AF 300 is a solution of an antifadent in glycerol and has been specially prepared for UVM-2 solutions where fluorescein is the staining material. The solutions may be used with other fluorochromes such as rhodamines etc. The presence of the antifadent helps to reduce photobleaching of the fluorophore. Solutions should be made up by mixing 1 part by volume of AF300 with 3 parts by volume of UVM-2. A few drops of the solution should be applied to the specimen followed by a cover slip. The mixture of AF300 and UVM-2 may be cured with light in the usual way to give a clear film. Application Sheet AF300 + UVM-2 (PDF 69KB) |
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CFPVOH
CFPVOH is an aqueous solution of poly(vinyl alcohol) for use as a permanent mountant. A few drops of the solution should be applied to the specimen followed by a cover slip. The water evaporates to give a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet CFPVOH (PDF 58KB) |
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CFPVOH plus antifadent
CFPVOH plus antifadent is an aqueous solution of poly(vinyl alcohol) containing an antifadent and is intended for use where it is desired to have a permanent mountant for a specimen which has been fluorescently labelled. The antifadent helps retard bleaching of fluorescent label with the extent of retardation being very dependent upon the type of label, and the molecular structure of the specimen. A few drops of the solution should be applied to the specimen followed by a coverslip. If the edges of the coverslip are sealed with nail varnish, the solution acts as an excellent gel mount. If the edges of the coverslip are not sealed water evaporates to leave a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. If bleaching of the label remains unacceptably high, try a mixture of CFPVOH plus antifadent with AF100 mixed in the ratio of 10:1 (v/v). This solution will have a short shelf life (a few days). Application Sheet CFPVOH + AF (PDF 59KB) |
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PVP
Aqueous poly(vinyl pyrrolidone) is very useful as a permanent mountant which possesses some antifadent properties. A few drops of the solution should be applied to the specimen followed by a cover slip. The water evaporates to give a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet PVP (PDF 58KB) |
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PVP + antifadent
Aqueous poly(vinyl pyrrolidone) plus antifadent is very useful as a permanent mountant which possesses antifadent properties. A few drops of the solution should be applied to the specimen followed by a cover slip. The water evaporates to give a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet PVP + AF (PDF 58KB) |
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Tris-MWL 4-88
Tris-MWL 4-88 solution is a solution of poly(vinyl alcohol)[Mowiol™ 4-88] in a water/glycerol/tris buffer mix and has a pH of 8.5. Following loss of water it forms good clear films and is therefore used as a permanent mountant. A few drops of the solution should be applied to the specimen followed by a cover slip. The water evaporates to give a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. If bleaching of the fluorescent marker is a problem, you may add AF100 as you would to CFPVOH (i.e.9 parts MWL solution plus 1 part AF100) These mixtures have a limited shelf-life. Application Sheet Tris-MWL (PDF 60KB) |
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CFM-1 *
This glycerol-phosphate buffered saline based mountant solution has
been specially formulated to have a refractive index of ~ 1.52 (at
room temperature) i.e. a refractive index similar to glass so that
effects of refraction on image quality are minimised. This mountant
solution may also be used as an immersion oil. The CFM solutions will
be particularly useful for high magnification work where immersion
oils are use to minimise distortion of the image due to refraction
of the viewing light. The solutions should also be particularly
valuable for lasing scanning microscopy where three-dimensional imaging
of specimens is carried out e.g. with confocal microscopy. The CFM-1 mountant/immersion oils should only be used with fixed i.e. cross-linked samples since linear biopolymers e.g. linear proteins, are solubilised by the mountant/immersion oils due to them containing a surfactant-type species Application Sheet CFM-1 (PDF 44KB) * This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application |
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CFM-1+ antifadent *
This glycerol-phosphate buffered saline based mountant solution has
been specially formulated so as to have a refractive index of ~ 1.52
(at room temperature) and contains an antifadent to retard the bleaching
of fluorochromes. Its refractive index is similar to glass and
therefore effects of refraction on image quality are minimised. The
solution may also be used as an immersion oil. The CFM-1 plus antifadent
solution will be particularly useful for high magnification work where
immersion oils are use to minimise distortion of the image due to refraction
of the viewing light and where bleaching of the fluorochrome occurs. The
solutions should also be very valuable for laser scanning microscopy
where three-dimensional imaging of specimens is carried out e.g. with
confocal fluorescence microscopy where integrity of the image has to
be maintained. The CFM-1 plus antifadent mountant/immersion oils should only be used with fixed i.e. cross-linked samples since linear biopolymers e.g. linear proteins, are solubilised by the mountant/immersion oils due to them containing a surfactant-type species Application Sheet CFM-1 + AF (PDF 44KB) *This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application |
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CFM-2 *
This glycerol- tris-buffered based mountant solution has been specially
formulated to have a refractive index of ~ 1.52 (at room temperature)
i.e. a refractive index similar to glass so that effects of refraction
on image quality are minimised. This mountant solution may also
be used as an immersion oil. The CFM solutions will be particularly
useful for high magnification work where immersion oils are use to
minimise distortion of the image due to refraction of the viewing light. The
solutions should also be particularly valuable for lasing scanning
microscopy where three-dimensional imaging of specimens is carried
out e.g. with confocal microscopy. The CFM-2 mountant/immersion oil should only be used with fixed i.e. cross-linked samples since linear biopolymers e.g. linear proteins, are solubilised by the mountant/immersion oils due to them containing a surfactant-type species Application Sheet CFM-2 (PDF 44KB) * This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application |
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CFM-3 *
This glycerol-phosphate based mountant solution containing
an antifadent has been specially formulated to have a refractive
index of ~ 1.52 (at room temperature) i.e. a refractive index similar
to glass so that effects of refraction on image quality are minimised. This
mountant solution may also be used as an immersion oil. The CFM solutions
will be particularly useful for high magnification work where immersion
oils are use to minimise distortion of the image due to refraction
of the viewing light. The solutions should also be particularly valuable
for lasing scanning microscopy where three-dimensional imaging of specimens
is carried out e.g. with confocal microscopy. The CFM-1 mountant/immersion oils should only be used with fixed i.e. cross-linked samples since linear biopolymers e.g. linear proteins, are solubilised by the mountant/immersion oils due to them containing a surfactant-type species Application Sheet CFM-3 (PDF 45KB) * This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application |
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Permafix-1
Permafix-1 is an aqueous solution of a chemically modified poly(vinyl alcohol) for use as a permanent mountant. The solution may be used with either AF100 or AF200 where it is important to reduce the photobleaching of fluorochromes (see Application Sheets for AF100 and AF200 for details). A few drops of the solution should be applied to the specimen followed by a cover slip. As the water evaporates crosslinking occurs which produces a clear hard robust film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet Permafix-1 (PDF 29KB) |
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Permafix-1(HRF)
Permafix-1 (HRF) is an aqueous solution of a chemically modified poly(vinyl alcohol) containing additives for use as a permanent mountant having a high refractive index (~1.52). The solution may be used with either AF100 or AF200 where it is important to reduce the photobleaching of fluorochromes (see Application Sheets for AF100 and AF200 for details) but dilution with these solutions will reduce the refractive index of the dried films. Permafix-1 HRF should only be used with specimens that have been fixed i.e. the biopolymers have been crosslinked. The additives in the formulation will dissolve linear proteins etc. leading to problems with obtaining good images. A few drops of the solution should be applied to the specimen followed by a cover slip. Evaporation of the water leads to formation of a clear film which holds the cover slip in place thereby aiding the safe storage of the specimen. Application Sheet Permafix-1(HRF) (PDF 29KB) |
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UVM-1
UVM-1 (a UV Mount medium) is a methacrylate-based solution which on exposure to light hardens to give clear film. It is totally water miscible and therefore can be used with non-dried specimens e.g. specimens that have been washed in buffer solutions. The light source may either be a desk lamp or sunlight! Since the solution cures to give a hard film there is no need to seal the outer edges of an applied coverslip with nail varnish etc. The preparation does not contain any volatile solvent but, as with all methacrylate solutions, care should be taken not to spill any of the material onto the skin (use appropriate gloves). If fading of fluorochromes is a problem, AF100 may be added (ratio of UVM-1 to AF100 being 10:1) Application Sheet UVM-1 (PDF 29KB) |
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UVM-2
UVM-2 (a UV Mount medium) is a methacrylate based solution containing an antibleaching agent, which on exposure to light hardens to give clear film. It is totally water miscible and therefore can be used with non-dried specimens e.g. specimens that have been washed in buffer solutions. The light source may either be a desk lamp or sunlight! Since the solution cures to give a hard film there is no need to seal the outer edges of an applied coverslip with nail varnish etc. The preparation does not contain any volatile solvent but, as with all methacrylate solutions, care should be taken not to spill any of the material onto the skin (use appropriate gloves). Application Sheet UVM-2 (PDF 29KB) |
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UVM-3
UVM-3 (a UV Mount medium) is a high refractive index methacrylate- based solution designed to be used with AF100 where fluorochrome fading is a problem. The solutions on exposure to light harden to give clear film. UVM-3 is totally water miscible and therefore can be used with non-dried specimens e.g. specimens that have been washed in buffer solutions. The light source may either be a desk lamp or sunlight! Since the solution cures to give a hard film there is no need to seal the outer edges of an applied coverslip with nail varnish etc. The preparation does not contain any volatile solvent but, as with all methacrylate solutions, care should be taken not to spill any of the material onto the skin (use appropriate gloves). It is important that the UVM-3 be only used with specimens that have been fixed i.e. the biopolymers have been crosslinked. The additives in the formulation will dissolve proteins etc. leading to problems with obtaining good images. Application Sheet UVM-3 (PDF 29KB) |
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