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In cell culture, a typical formaldehyde fixing procedure would involve using a 4% formaldehyde solution in phosphate buffered saline (PBS) on ice for 10 minutes. In histology and pathology specimens preparation, usually, the fixation step is performed using 10% Neutral Buffered Formalin (4% formaldehyde) for, at least, 24 hours.
The most commonly used fixative in histology is formaldehyde. It is usually used as a 10% neutral buffered formalin (NBF), that is approx. 3.7%–4.0% formaldehyde in phosphate buffer, pH 7. Since formaldehyde is a gas at room temperature, formalin – formaldehyde gas dissolved in water (~37% w/v) – is used when making the former fixative.
It was invented by French biologist Pol Bouin and is composed of picric acid, acetic acid and formaldehyde in an aqueous solution. [2] Bouin's fluid is especially useful for fixation of gastrointestinal tract biopsies because this fixative allows crisper and better nuclear staining than 10% neutral-buffered formalin.
If the PAS stain will be performed on tissue, the recommended fixative is 10% neutral-buffered formalin or Bouin solution. For blood smears, the recommended fixative is methanol. Glutaraldehyde is not recommended because free aldehyde groups may be available to react with the Schiff reagent, which may result in false positive staining. [4]
If the glacial acetic acid is replaced by 5 ml of formalin (37–40% formaldehyde), the resulting solution is Helly's fixative, also sometimes called "formol-Zenker".Helly is stable for only a few hours because the formaldehyde and dichromate components react, producing formic acid and chromium(III) ions; the orange solution becomes greenish.
Neutral red (toluylene red, Basic Red 5, or C.I. 50040) is a eurhodin dye used for staining in histology. It stains lysosomes red. [ 1 ] It is used as a general stain in histology, as a counterstain in combination with other dyes, and for many staining methods.
Buffer capacity falls to 33% of the maximum value at pH = pK a ± 1, to 10% at pH = pK a ± 1.5 and to 1% at pH = pK a ± 2. For this reason the most useful range is approximately p K a ± 1. When choosing a buffer for use at a specific pH, it should have a p K a value as close as possible to that pH.
Buffered oxide etch (BOE), also known as buffered HF or BHF, is a wet etchant used in microfabrication. It is a mixture of a buffering agent, such as ammonium fluoride NH 4 F, and hydrofluoric acid (HF). Its primary use is in etching thin films of silicon nitride (Si 3 N 4) or silicon dioxide (SiO 2), by the reaction: SiO 2 + 4HF + 2NH 4 F → ...
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