Home LiteratureArticle Details
PMID: 825524 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Improved procedures for immunoferritin labeling of ultrathin frozen sections.

The Journal of cell biology ·Vol. 71 ·No. 3 ·1976-12-00 ·Pages 894-906

Tokuyasu KT, Singer SJ

Abstract

In employing fixed frozen ultrathin sections as substrates for immunoferritin labeling of intracellular antigens, we have found that conventional glutaraldehyde fixation sometimes permits very little specific staining of the sections, either because it inactivates certain protein antigens, or because it renders them inaccessible to the antibody stains. We have developed several fixation procedures that are chemically milder and allow a uniform but less extensive cross-linking of the specimen. With these procedures and precautions in the handling of the more fragile frozen sections, excellent structural preservation and specific immunoferritin labeling has been achieved with several systems.

MeSH Terms
Antigens/analysis Erythrocytes/analysis Fixatives Fluorescent Antibody Technique Formaldehyde Frozen Sections Glutaral Microtomy Muscle, Smooth/analysis Muscles/analysis Specimen Handling
Chemicals
Antigens Fixatives Formaldehyde Glutaral
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tokuyasu K T
Singer S J
References (9)
9 references, click to expand
  1. A technique for ultracryotomy of cell suspensions and tissues.
    J Cell Biol. 1973 May;57(2):551-65 PMID: 4121290
  2. Immunoferritin localization of intracellular antigens: the use of ultracryotomy to obtain ultrathin sections suitable for direct immunoferritin staining.
    Proc Natl Acad Sci U S A. 1973 Jun;70(6):1649-53 PMID: 4124304
  3. Anionic sites of human erythrocyte membranes. II. Antispectrin-induced transmembrane aggregation of the binding sites for positively charged colloidal particles.
    J Cell Biol. 1973 Nov;59(2 Pt 1):395-406 PMID: 4141707
  4. Bifunctional imidoesters as cross-linking reagents.
    Biochem Biophys Res Commun. 1966 Jun 13;23(5):730-9 PMID: 4164335
  5. Membranes as observed in frozen sections.
    J Ultrastruct Res. 1976 May;55(2):281-7 PMID: 58070
  6. A disulfide-bridge bifunctional imidoester as a reversible cross-linking reagent.
    Biochem Biophys Res Commun. 1975 Mar 17;63(2):417-23 PMID: 1168466
  7. Tissue fixation with diimidoesters as an alternative to aldehydes. I. Comparison of cross-linking and ultrastructure obtained with dimethylsuberimidate and glutaraldehyde.
    J Histochem Cytochem. 1974 Apr;22(4):223-9 PMID: 4208871
  8. Two improved methods for preparing ferritin-protein conjugates for electron microscopy.
    J Cell Biol. 1975 Feb;64(2):331-9 PMID: 803973
  9. Alteration of the conformation of proteins in red blood cell membranes and in solution by fixatives used in electron microscopy.
    J Cell Biol. 1968 Apr;37(1):117-21 PMID: 5645836
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1976-12-00
Pages
894-906
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2109786
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]