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Journal of Histochemistry and Cytochemistry
Volume 52 (1): 141-144, 2004
Copyright ©The Histochemical Society, Inc.

BRIEF REPORT

Post-embedding Double-labeling of Antigen-retrieved Ultrathin Sections Using a Silver Enhancement-controlled Sequential Immunogold (SECSI) Technique

Nigel P. Goode, Michael Shires, Doreen M. Crellin, Tahir N. Khan and Andrew F. Mooney

Renal Research Unit (NPG,MS,DMC) and Departments of Histopathology (TNK) and of Renal Medicine (AFM), St James's University Hospital, The Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom

Correspondence to: Dr. N.P. Goode, Renal Research Unit, Clinical Sciences Building, St. James's University Hospital, Leeds LS9 7TF, UK. E-mail: n.p.goode{at}leeds.ac.uk

In electron microscopy, the post-embedding immunogold technique provides a high degree of resolution and the possibility of quantitation owing to the intrinsic characteristics of the colloidal gold marker. Application of this technique to the subcellular localization of multiple antigens by differential labeling using gold markers of different sizes, or to double labeling using the same primary antibody isotype with serial silver enhancement, has been reported. We have incorporated this double labeling technique into a modified procedure that produces excellent labeling and ultrastructural preservation, even after exposure of ultrathin sections large enough to cover a 300-µm-diameter single-hole grid to hot antigen retrieval solutions and prolonged labeling protocols.

(J Histochem Cytochem 52:141–144, 2004)

Key Words: immunogold • silver enhancement • antigen retrieval • post-embedding labeling • electron microscopy • LR Gold


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Mechanisms of Heat-induced Antigen Retrieval: Does pH or Ionic Strength of the Solution Play a Role for Refolding Antigens?
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[Abstract] [Full Text] [PDF]



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