Increased yield of immunogold labeling of epoxy sections by adding para-phenylendiamine in the tissue processing

Brorson, Sverre‐Henning; Halvorsen, I.; Lønning, L.-C.; Slaattun, G.; Sletten, Mira Aaboen; Rashid, Shannon K.

doi:10.1016/s0968-4328(99)00058-x

Cited by 9 publications

(1 citation statement)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…p -Phenylenediamine (PPD) has been used to retard photobleaching in fluorescence microscopy, to increase the intensity of osmium staining [1, 2], and to improve lipid preservation [3, 4]. Phend et al [5] used PPD to improve post-embedding immunolocalization in epoxy-embedded specimens; more recently, Brorson et al [6] recommended including 1% (wt/vol) PPD in all dehydration steps for post-embedding immunolocalization. Experience with a number of animal tissues and microorganisms has resulted in the following protocol, which has been used to improve post-embedding immunolocalization in specimens embedded in either epoxy resins or acrylic resins such as LR White and LR Gold.…”

mentioning

confidence: 99%

p-Phenylenediamine: An Adjunct to and a Substitute for Osmium Tetroxide

Ellis

2010

Micros. Today

View full text Add to dashboard Cite

p-Phenylenediamine (PPD) has been used to retard photobleaching in fluorescence microscopy, to increase the intensity of osmium staining , and to improve lipid preservation . Phend et al. used PPD to improve post-embedding immunolocalization in epoxy-embedded specimens; more recently, Brorson et al. recommended including 1% (wt/vol) PPD in all dehydration steps for post-embedding immunolocalization. Experience with a number of animal tissues and microorganisms has resulted in the following protocol, which has been used to improve post-embedding immunolocalization in specimens embedded in either epoxy resins or acrylic resins such as LR White and LR Gold.

show abstract

mentioning

confidence: 99%

p-Phenylenediamine: An Adjunct to and a Substitute for Osmium Tetroxide

Ellis

2010

Micros. Today

View full text Add to dashboard Cite

show abstract

Immunocytochemistry

Dykstra

Reuss

2003

Biological Electron Microscopy

View full text Add to dashboard Cite

Staining Sectioned Biological Specimens for Transmission Electron Microscopy: Conventional and En Bloc Stains

Ellis

2013

Methods in Molecular Biology

View full text Add to dashboard Cite

Post-staining of ultrathin sections and/or en bloc staining of specimens is necessary for differential contrast and improved resolution of cellular structures. Often specimens are fixed and stained with osmium tetroxide during fixation, but additional contrast is the result of additional heavy metal stains on the sections. The most common post-staining of sections is done on grids by aqueous uranyl acetate followed by lead citrate. When it is apparent that simple, aqueous uranium and lead post-staining is not adequate, other stains are invoked. These procedures can be as simple as en bloc staining with uranyl acetate after primary fixation and osmication. Over the years, several other treatments have been developed for use with the primary fixation or during dehydration. Tannic acid, paraphenylenediamine (PPD), and malachite green can all serve as en bloc stains and can contribute to overall improved visualization of ultrastructural details in biological specimens. Tannic acid and PPD improve membrane preservation, and malachite green is a phospholipid stain. All of these stains are compatible with aqueous fixatives and should be considered when the usual stains are not satisfactory. Marinozzi rings and microwave-assisted post-staining offer alternatives to traditional grid staining. In addition, stain precipitates on grids often can be removed by treatment with 10 % (v/v) acetic acid.

show abstract

Increased yield of immunogold labeling of epoxy sections by adding para-phenylendiamine in the tissue processing

Cited by 9 publications

References 13 publications

p-Phenylenediamine: An Adjunct to and a Substitute for Osmium Tetroxide

p-Phenylenediamine: An Adjunct to and a Substitute for Osmium Tetroxide

Immunocytochemistry

Staining Sectioned Biological Specimens for Transmission Electron Microscopy: Conventional and En Bloc Stains

Contact Info

Product

Resources

About