Temperature, Peroxide Concentration, and Immunohistochemical Staining Method Affects Staining Intensity, Distribution, and Background

Development of Ig-synthesizing cells in the bursa of chick embryo was studied by immunohistochemical staining method as well as by in vitro incorporation of leucine-3H into Ig. Ig-synthesizing cells are first detected in the bursa of a 14 day old chick embryo and increase with the maturation of the embryo. Acrylamide gel analysis of leucine-3H-labeled Ig shows that synthesis of nonsecretory IgM-H0 precedes that of secretory IgM-H, reflecting an ontogenetic sequence of development of lymphoid cells synthesizing IgM. Since IgM-H0 is not secreted, we further studied biochemical differences between two heavy chains. The difference is attributable to lack of galactose attachment to H0 chains. It is proposed that during differentiation of lymphoid cells synthesizing and secreting Ig, attachment of galactose may play an obligatory role in the development of cellular capacity for Ig secretion.

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Double Immunohistochemical Staining Method for HIF-1α and its Regulators PHD2 and PHD3 in Formalin-fixed Paraffin-embedded Tissues

Applied Immunohistochemistry & Molecular Morphology ◽

10.1097/pai.0b013e3181d6bd59 ◽

2010 ◽

Vol 18 (4) ◽

pp. 375-381 ◽

Cited By ~ 6

Author(s):

Mary M. Vaughan ◽

Karoly Toth ◽

Sreenivasulu Chintala ◽

Youcef M. Rustum

Keyword(s):

Immunohistochemical Staining ◽

Staining Method ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Double Immunohistochemical Staining ◽

Formalin Fixed

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A Novel Immunohistochemical Staining Method Allows Ultrarapid Detection of Lymph Node Micrometastases While Conserving Antibody

ACTA HISTOCHEMICA ET CYTOCHEMICA ◽

10.1267/ahc.11006 ◽

2011 ◽

Vol 44 (3) ◽

pp. 133-139 ◽

Cited By ~ 20

Author(s):

Hiroshi Toda ◽

Yoshihiro Minamiya ◽

Masami Kagaya ◽

Hiroshi Nanjo ◽

Yoichi Akagami ◽

...

Keyword(s):

Lymph Node ◽

Immunohistochemical Staining ◽

Staining Method ◽

Lymph Node Micrometastases

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Quantitative Comparison of Immunohistochemical Staining Intensity in Tissues Fixed in Formalin and Histochoice

Analytical Cellular Pathology ◽

10.1155/1997/607965 ◽

1997 ◽

Vol 15 (2) ◽

pp. 119-129 ◽

Cited By ~ 18

Author(s):

D. Geoffrey Vince ◽

Abdelghani Tbakhi ◽

Ajeetkumar Gaddipati ◽

Robert M. Cothren ◽

J. Fredrick Cornhill ◽

...

Keyword(s):

Immunohistochemical Staining ◽

Staining Intensity ◽

Quantitative Comparison ◽

Color Images ◽

Cross Linking ◽

Fixed Tissue ◽

Tissue Samples ◽

Significant Difference ◽

Immunohistochemical Stains

Formaldehyde fixatives have traditionally been used to preserve tissues as they impart excellent morphological preservation. Formaldehyde fixes tissue by cross linking, a process which can reduce the antigenicity of tissue and weakens the intensity of immunohistochemical stains. Preliminary studies have shown that Histochoice tissue fixative offers equal or greater staining intensity than neutral buffered formalin (NBF). This study compares these fixatives quantitatively and presents the results in unambiguous statistical terms.Tissue samples were collected, bisected, and fixed in NBF or Histochoice. The sections were stained with a total of 21 antibodies, and color images were collected. The hue, saturation, and value were determined for each positive pixel and an ANOVA performed. Small differences in hue were noted in 8 of 21 cases. Histochoice‐fixed tissue gave a greater mean saturation than NBF with 57.1% of the antibodies tested. No significant difference in the saturation was detected in 28.6% of the cases; NBF gave higher mean saturation levels with only 14.3% of the antibodies. Histochoice‐fixed tissue was found to give lower values in 66.7% of cases than those prepared with NBF, indicating darker staining. These results show that Histochoice produces staining intensity that is comparable, and in many cases superior, to formalin.

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