Anti fluorescence decay sealing agent Citifluor ™

A series of products specially designed for immunofluorescence reaction encapsulation, with unique formulas that prevent the fluorescence of dyes from fading.Common anti fluorescence decay sealing agents includeAF1TheAF2TheAF3TheAF4TheAF87TheAF100TheAF200TheAF300TheCFM-1TheCFM-2wait. Some of the products includeDAPI,Can stain the nucleus or stain the nucleusDNAThe experiment.

Regarding anti fluorescence quenchers, there are three common types: phenolic propyl gallaten-Propyl gallate (NPG)Amine type p-phenylenediaminep-Phenylenediamine (PPD)Amine type1,4-Diazabicyclo-octane（DABCO）Triethylenediamine. The basic theory of fluorescence quenching is as follows: ordinary dissolved oxygen in tissue slices becomes singlet oxygen when excited, which is similar to oxygen free radicals. This is the main culprit of fluorescence quenching, and the anti quenching agent we add mainly reacts with this culprit to directly remove singlet oxygen. Note that adding an anti quenching agent to the sealing agent often reduces the total fluorescence in the primary stage of excitation, and alsoDNARisks associated with dye reactions. Take a look at the advantages and disadvantages of these three reagents:PPDCan react with cyanine dyes (especiallyCy2）Therefore, this type of dye sealing film is not considered to containPPDThe sealing agent;NPGNon toxic, this type of sealing agent is preferred for fluorescent staining of live cells, butNPGInterference with the process of cell apoptosis, students conducting such research should pay more attention and try to avoid containingNPGThe sealing agent;DABCOunlikePPDThat is effective and has low toxicity, and can be used for live cells, but it isNPGSimilarly, it interferes with the process of cell apoptosis.

How to quickly find the one you need with a large amount of sealant piled up here? The simple idea is:

The first step is solidification or non solidification? Solidification makes storage and retrieval very convenient, but it is important to consider the shrinkage of tissue pieces during the solidification process and the shrinkage that requires dehydration steps. Non solidified type requires consideration of the evaporation and side leakage of the sealing agent, and of course, a cover glass (sometimes also requiring a silicone sealing gasket) is neededspacer）And often the cover glass needs to be sealed, usually nail polish, dental waxVALAP (equal mixture of Vaseline, lanolin, and paraffin mixed on a heating plate 60°C)Apply it around the cover glass. It takes a long time for nail polish to dry, which may pollute the lens; Dental wax can solidify quickly when removed at room temperature;VALAP (equal mixture of Vaseline, lanolin, and paraffin mixed on a heating plate 60°C)It is commonly used in live cell sealing.

Step 2: Do you need glycerols (water-soluble) or non glycerols (lipophilic)? Glycerol can be miscible with any proportion of water, can be miscible with ethanol, but is not miscible with oil. Knowing these makes the choice much simpler. What is your chemical reaction system like? If it contains water, choose glycerol based (water-soluble) ones. After strict dehydration treatment, non glycerol based (oil soluble) is selected.

Step three, the refractive index of the sealing agent. The refractive index of the stained film is selected using a sealing agent1．52to1．54The best transparency between them; It is best to use a sealing agent with high or low refractive index to seal unstained tissues, which can eliminate spherical aberration and obtain the best image.

Introduce the film-forming agents used:

polyvinyl alcoholpoly (vinyl alcohol)Organic polymer substance, non-toxic, soluble in water, commonly used as a film-forming agent. Assign different numbers based on their degree of polymerization and alcoholysis, for exampleMusic ® 4-88.PVAOther substances or reactions can also be added for modification treatment, such asPermafix-1It's modifiedPVA.

Polyvinylpyrrolidone(polyvinyl pyrrolidone)abbreviationPVP, is a non-ionic polymer compound and a commonly used film-forming agent.

Product selection guide:

l AF1:The basic ingredients are glycerolPBSBuffer solution, anti fluorescence quencher, other components, refractive index, etc1.46，PH10Left and right, suitable for detecting tissue slices, dead cells, etc. soaked in glycerol FISH (including CARD-FISH)Waiting for the film.

l AF2:The basic ingredients are glycerol, anti fluorescence quencher, other components, etc., without its own buffer solution. refractive index1.47The advantage of this product is that it does not have its own buffer and can be compatible with any other buffer.

l AF3:The basic components arePBSBuffer solution, anti fluorescence quencher, other components, refractive index1.34Especially suitable for live cell testing.

l AFR3:The basic components arePBSBuffer solution, a latest type of anti fluorescence quencher (non amine non phenolic), other ingredients, etc.

l AF4:also known asGlycerol/n-propyl gallate mounting mediumSealing agent, the added anti fluorescence decay agent is propyl gallaten-Propyl gallate (NPG).

l AF87The special feature is that it contains non fluorescent immersion oil, which is a specially designed sealing agent for observation under high magnificationDAPIDyeing is particularly useful（DAPIThere is a tendency for dispersion in glycerol based tablets.

l AF100:The basic components arePBSBuffer solution, anti fluorescence decay agent, other ingredients, etc. This is a glycerol free sealing agent that needs to be combined withCFPVOHUsed in conjunction with the solution. Pre mixing is requiredAF100andCFPVOHSolution, mix as much as needed, after mixing10Used up within hours. The refractive index of the sealing agent1.34-1.35.

l AF200:andAF100Similar, but different,AF200It's glycerol based,Amino based anti fluorescence decay agent, suitable for packaging anthocyanins, Alexa, Hoeches, and other products.

l AF300:andAF100Similar, but different,AF300It is a glycerol based, phenolic based anti fluorescence decay agent.

l CFMThe refractive index of the series of sealing agents is1.52Equal to or approximately equal to the refractive index of the carrier and cover glass used, which is very helpful for obtaining perfect images. Mainly used under laser confocal microscopy or as a substitute for immersion in oil for high-power mirrors.CFM-1ofpHabout6.5，.CFM-1 Plus AntifadingofPHThe value is approximately10，CFM-2ofpH ofabout~8.5，CFM-3ofPHThe value is close to neutral.

Ordering information: