Citifluor Ltd - Antifadent Solutions Citifluor Ltd - Antifadent Solutions

 

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AF1

Properties and storage of AF1 mountant solutions
The solutions are of medium viscosity and are water-white in appearance. They may be stored at room temperature and ideally between 5° and 15° and out of strong sunlight. The cap of the bottle or if using the pipette supplied with the material, the cap which covers the pipette delivery point, should always be replaced after use as a matter of good practice. Samples stored under these conditions for 6 months have shown no apparent deterioration. If the AF1 solution is being used in an assay, a control experiment should always be carried out.

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.

viscosity

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)

AF2

Properties and storage of AF2 mountant solutions
The solutions are of medium viscosity and are water-white in appearance. They may be stored at room temperature and ideally between 5° and 15° and out of strong sunlight. The cap of the bottle or if using the pipette supplied with the material, the cap which covers the pipette delivery point, should always be replaced after use as a matter of good practice and also to prevent the ingress of water (due to the glycerol being hygroscopic). Samples stored under these conditions for 6 months have shown no apparent deterioration. If the AF2 solution is being used in an assay, a control experiment should always be carried out.

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)

AF3

Properties and storage of AF3 mountant solutions
The solutions have a water-white in appearance. They may be stored at room temperature and ideally between 5° and 15° and out of strong sunlight. The cap of the bottle or if using the pipette supplied with the material, the cap which covers the pipette delivery point, should always be replaced after use as a matter of good practice. Samples stored under these conditions for 6 months have shown no apparent deterioration. If the AF3 solution is being used in an assay, a control experiment should always be carried out.

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)

AF87

Properties and storage of AF87
The immersion oil may be stored at 5 to 10°C and should be kept out of strong sunlight. It should not be stored in a refrigerator. Occasionally small crystals are formed in the oil but careful use of a pipette can obviate the crystals interfering with the function of the oil

AF87 is a non-fluorescent immersion oil of medium viscosity and has a refractive index of 1.516, and should be used in the usual way. It is recommended that the microscope objective is wiped clan after use. A useful solvent mixture for removing the immersion oil is composed of ether /ethanol(??/?? v/v). The oil contains an antifadent for those who wish to use it as a mountant solution. The new CFM series of mountant solutions is designed is designed to be used both as a mountant solution and as an immersion oil,

Application Sheet AF87 (PDF 30KB)

AF100 + CFPVOH

Properties and storage of CFPVOH solutions containing AF100
The solutions are of medium viscosity and are water-white in appearance. The CFPVOH solution contains a small amount of sodium azide to prevent fungal growth. The AF100/CFPVOH solution does not keep and the efficacy of the antifadent is markedly reduced over a 10 hour period. With this in mind, it is better to make up the mixture prior to use and not to rely on keeping solutions

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)

CFPVOH

Properties and storage of CFPVOH solutions
The solutions are of medium viscosity and are water-white in appearance. They contain a small amount of sodium azide to prevent fungal growth. They may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water which leads to film formation and ultimately to solidification of the product. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration e.g. no gel formation.

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)

CFPVOH plus antifadent

Properties and storage of CFPVOH plus antifadent solutions
The solutions are of medium viscosity, are water-white in appearance and have a pH of ~8.5. They contain a small amount of sodium azide to prevent fungal growth. They may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water which leads to film formation and ultimately to solidification of the product. Samples stored under these conditions for 2 months have been found to exhibit little apparent deterioration e.g. no gel formation.

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 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 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)

PVP

Properties and storage of Poly(vinyl pyrrolidone) solutions
The solutions are of medium viscosity and are slightly yellow in appearance. They have a pH of ~7. If you require a higher pH you may add a buffer solution such as a tris-solution but this may affect the shelf-life of the solution. The solutions may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water which leads to film formation and ultimately to solidification of the product. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration e.g. no gel formation.

If bleaching of your fluorescent label is a problem use the poly(vinyl pyrrolidone) solution containing antifadent.

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)

PVP + antifadent

Properties and storage of Poly(vinyl pyrrolidone) plus antifadent solutions
The solutions are of medium viscosity and are slightly yellow in appearance. They have a pH of ~9.5. The solutions may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water which leads to film formation and ultimately to solidification of the product. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration e.g. no gel formation.

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)

Tris-MWL 4-88

Properties and storage of MWL solutions
The solutions are of medium viscosity, are water-white in appearance and have a pH of ~8.5. They may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water which leads to film formation and ultimately to solidification of the product. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration e.g. no gel formation.

If you would prefer to have a solution of higher viscosity we can make these to order.

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)

CFM-1 *

Properties and storage of CFM solutions
The solution is of medium viscosity, is water-white in appearance and has a pH of ~6.5. The solution may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration.

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.

A few drops of the solution should be applied to the specimen followed by a cover slip. An immersion oil (which may be CFM solution if its viscosity is appropriate) is applied in the usual way.

Application Sheet CFM-1 (PDF 56KB)

* This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application

CFM-1+ antifadent *

Properties and storage of CFM 1 solutions plus antifadent
The solutions are of medium viscosity, are water-white in appearance and have a pH of ~10. The solution may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration.

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 contain 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.

A few drops of the solution should be applied to the specimen followed by a cover slip. An immersion oil (which may be CFM-1 solution if its viscosity is appropriate) is applied in the usual way.

Application Sheet CFM-1 + AF (PDF 56KB)

* This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application

CFM-2 *

Properties and storage of CFM solutions
The solution is of medium viscosity, is water-white in appearance and has a pH of ~8.5 in order to maximise the signals from FITC labelled specimens. The solution may be stored at room temperature. The cap of the bottles should always be replaced after use, to prevent evaporation of water. Samples stored under these conditions for 6 months have been found to exhibit little apparent deterioration.

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.

A few drops of the solution should be applied to the specimen followed by a cover slip. An immersion oil (which may be CFM solution if its viscosity is appropriate) is applied in the usual way.

Application Sheet CFM-2 (PDF 56KB)

* This and related high refractive index dual mountant solutions and immersion oils are the subject of a current patent application

 

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