Questions? 800-523-5874 | [email protected]
Mountant Media Containing Antifadents – The Antidote For Photobleaching
Antifadent Solutions reduce the photo-bleaching, or fading of the fluorescence of dyes used for labeling biological species. The fading of fluorochrome dyes is a particular problem in fluorescence microscopy, such as in immunofluorescence studies. Fluorescent dyes are also used as cell markers, for following the uptake and release of calcium from cells for example, and for characterizing cell surfaces. For work at high magnification, a non-fluorescent immersion oil is available.
In fluorescence microscopy, the fluorescence is stimulated by high intensity UV or visible light. Absorption of the light populates an excited state of the dye (usually a singlet state) and this leads to fluorescence. However, the excited state of the dye may undergo chemical reactions which leads to its destruction as evidenced by the fading or bleaching of the fluorescence and consequent loss of the image or in the case of assays, a change in signal intensity during measurement.
To overcome these problems, use antifadent (antibleaching) solutions as mounting media when specimens are examined by fluorescent microscopy or other detection systems and as additives for assays.
The most frequently used solution is AF1 which is an anti-fadent (anti-bleaching agent) contained in a glycerol PBS (phosphate-buffered saline) solution and is particularly useful for examining tissue sections.
AF2 solution contains the antifadent in glycerol which enables users to choose their own buffer, and AF3 is the antifadent in PBS solution. AF3 is particularly useful for examining live cells.
AF87 is a non fluorescent immersion oil containing antifadent. CFPVOH is an aqueous solution of poly (vinyl alcohol) for use as a solid mountant. AF100 is a solution of antifadents (anti bleaching agents) for use with CFPVOH when fading (bleaching) of specimens in a solid mountant is a problem.
None of the antifadents is based on p-phenylenediamine. All solutions may be stored at room temperature.
The table below illustrates the main types of antifadent available.
A more detailed account of our anti-fadent products can be found in the Technical Data Sheets below. The following is a brief overview which we hope helps you to decide on the best solution for your application.
If you are still unsure about which mountant to use, please don't hesitate to contact us, either by phone or email. We have technical staff on hand who are happy to answer the simplest, or the most challenging of questions.
These solutions have a refractive index that matches that of the glass of the coverslip thereby minimising the effects of spherical aberration. Such mountants are invaluable for imaging the internal structures of samples by techniques such as single muylti-photon confocal niche since it acts as an efficient Cleaing Solution (clearing occurs within a few minutes – Sean Speese, Oregon Health and State University, Personal Communication). The beneficial effects of these reagents can be seen in the two figures below where it can be seen that high resolution images have been obtained at a significant depth within the mountant solution. Additional features of these solutions includes their total water miscibility, they are non-odorous and do not quench the fluorescence of fluorochromes (unlike thiodiethanol, another proposed high refractive index mountant) and the solutions are stable for long periods (at least 1 year).
AF87 is an immersion oil having a refractive index of 1.52 and contains an antifadent. It may be used as an immersion oil and also as a mountant. Since AF87 is an oil that is immiscible with water, it is essential that specimens are dehydrated before application of the mountant.
Hardening mountants are aqueous solutions containing a polymer such a poly (vinyl alcohol) (PVOH). When a few microlitres of these solutions are pipetted onto a microscope slide and a coverslip applied, evaporation of water slowly takes place and the formation of a stable film results which immobilises the coverslip.
A variety of hardening mountants is available which provide films possessing a range of hardness. They are based on water-soluble polymers such as poly(vinyl alcohol(PVOH) and poly (vinyl pyrrolidone)(PVP).
PVOH is produced by hydrolysis of poly(vinyl acetate) and most commercial samples of PVOH contain residual (unhydrolysed) acetate groups. These groups undergo hydrolysis, often accelerated by the added antifadents, during storage and this can cause a change in pH and more usually gellation.
This latter process leads to an unpredictable shelf-life. By making up small volumes of PVOH solution containing the required amount of antifadent solution, you have materials of consistent composition and performance as well as making better use of your purchased materials.
Cat # | Description | MSDS | Pack | Price | Quote | Quantity | |
---|---|---|---|---|---|---|---|
Cat #: 17970-25 | Description: Citifluor Af1 | MSDS | Pack: 25 mL | Price: $43.50 |
Add to Quote:
|
|
|
Cat #: 17970-100 | Description: Citifluor Af1 | MSDS | Pack: 100 mL | Price: $114.50 |
Add to Quote:
|
|
|
Cat #: 17971-25 | Description: Citifluor Af2 | MSDS | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17971-100 | Description: Citifluor Af2 | MSDS | Pack: 100 mL | Price: $114.50 |
Add to Quote:
|
|
|
Cat #: 17972-25 | Description: Citifluor Af3 | MSDS | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17972-100 | Description: Citifluor Af3 | MSDS | Pack: 100 mL | Price: $114.50 |
Add to Quote:
|
|
|
Cat #: 17976-10 | Description: Citifluor Af87 | MSDS | Pack: 10 mL | Price: $39.00 |
Add to Quote:
|
|
|
Cat #: 17976-25 | Description: Citifluor Af87 | MSDS | Pack: 25 mL | Price: $63.50 |
Add to Quote:
|
|
|
Cat #: CFP-25 | Description: Citifluor Cfpvoh | MSDS | Pack: 25 mL | Price: $42.50 |
Add to Quote:
|
|
|
Cat #: AF100-5 | Description: Citifluor Af100 | MSDS | Pack: 5 mL | Price: $12.50 |
Add to Quote:
|
|
|
Cat #: 17978-35 | Description: Citifluor Cfpvoh + Af100* | MSDS | Pack: 30 mL | Price: $49.00 |
Add to Quote:
|
|
|
Cat #: 17980-10 | Description: Citifluor Cfm-1 | MSDS | Pack: 10 mL | Price: $36.00 |
Add to Quote:
|
|
|
Cat #: 17980-25 | Description: Citifluor Cfm-1 | MSDS | Pack: 25 mL | Price: $59.50 |
Add to Quote:
|
|
|
Cat #: 17981-10 | Description: Cfm-1 + Antifadent | MSDS | Pack: 10 mL | Price: $37.00 |
Add to Quote:
|
|
|
Cat #: 17981-25 | Description: Cfm-1 + Antifadent | MSDS | Pack: 25 mL | Price: $65.50 |
Add to Quote:
|
|
|
Cat #: 17982-10 | Description: Citifluor Cfm-2 | MSDS | Pack: 10 mL | Price: $36.00 |
Add to Quote:
|
|
|
Cat #: 17982-25 | Description: Citifluor Cfm-2 | MSDS | Pack: 25 mL | Price: $58.50 |
Add to Quote:
|
|
|
Cat #: 17970-125 | Description: Citifluor Af1 + Dapi | MSDS | Pack: 15 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17973-10 | Description: Citifluor Afr3 | MSDS | Pack: 10 mL | Price: $68.00 |
Add to Quote:
|
|
|
Cat #: 17973-25 | Description: Citifluor Af4 | MSDS | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17977-10 | Description: Citifluor Af200 | MSDS | Pack: 10 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17977-25 | Description: Citifluor Af300 | MSDS | Pack: 10 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17977-50 | Description: Citifluor Pvp-1 | MSDS | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17977-100 | Description: Citifluor Pvp-1 + Antifadent | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
||
Cat #: 17977-150 | Description: Citifluor Mwl4-88 | MSDS | Pack: 25 mL | Price: $44.50 |
Add to Quote:
|
|
|
Cat #: 17979-10 | Description: Citifluor Cfmr2 | MSDS | Pack: 10 mL | Price: $52.00 |
Add to Quote:
|
|
|
Cat #: 17979-20 | Description: Citifluor Cfm-3 | MSDS | Pack: 10 mL | Price: $37.00 |
Add to Quote:
|
|
|
Cat #: 17979-30 | Description: Citifluor Cfm-3 | MSDS | Pack: 25 mL | Price: $62.50 |
Add to Quote:
|
|