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Analytical Cellular Pathology
Volume 15, Issue 2, Pages 119-129

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

D. Geoffrey Vince,1 Abdelghani Tbakhi,2 Ajeetkumar Gaddipati,1,3 Robert M. Cothren,1 J. Fredrick Cornhill,1 and Raymond R. Tubbs2

1Whitaker Biomedical Imaging Laboratory, Department of Biomedical Engineering, Ohio, USA
2Department of Clinical Pathology, The Cleveland Clinic Foundation, Cleveland, Ohio, USA
3Biomedical Engineering Center, The Ohio State University, Columbus, Ohio, USA

Received 10 March 1997; Revised 7 June 1997; Accepted 10 June 1997

Copyright © 1997 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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.