Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-cd9895bd7-q99xh Total loading time: 0 Render date: 2024-12-26T15:47:39.972Z Has data issue: false hasContentIssue false

Chapter 7 - Troubleshooting Immunohistochemistry

Published online by Cambridge University Press:  16 June 2022

By
David Gan
Edited by
Trung Nguyen
Trung Nguyen
Affiliation:
Peter MacCallum Cancer Centre, Australia
Get access

Summary

Immunohistochemistry assays can be technically difficult. There are countless opportunities for errors. Some faults can be fixed, and this chapter contains a precise appraisal of common problems encountered along with suggested solutions. Incorporating both pre-analytic conundrums and analytic challenges from a staining perspective, it is written by a medical scientist able to think outside the square and offer resolutions ranging from the practical to the unconventional. All prescribed advice has been tested and proven successful. Suggesting corrective actions is an exercise in giving the worker options and workarounds for certain situations. Doing more with less and less testing material and lifting sections from slides to be used again when the paraffin block is exhausted are two examples of what can be achieved. Ultimately, it is about tuning protocols to make the most of the precious test tissue given whilst eliminating unnecessary wastage.

Keywords

lifting sectionsartefacttechnical difficultysolutionpracticalconundrumworkaroundprotocol
Type
Chapter
Information
Immunohistochemistry
A Technical Guide to Current Practices
 , pp. 183 - 209
Publisher: Cambridge University Press
Print publication year: 2022

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Athanasou, NA, Quinn, JMW, Heryet, A, Woods, CG, Mcgee, JO 1987. Effect of decalcification agents on immunoreactivity of cellular antigens. J Clin Pathol 40(8):874–8.CrossRefGoogle ScholarPubMed
Boenisch, T, Farmilo, AJ, Stead, RH, Key, M, Welcher, R, Harvey, R, Atwood, KN, comps. Handbook on immunohistochemical staining methods. 3rd edition. DAKO; 2001.Google Scholar
Bussolati, G, Leonardo, E 2008. Technical pitfalls potentially affecting diagnoses in immunohistochemistry. J Clin Pathol 61(11):1184–92.CrossRefGoogle ScholarPubMed
CAP 2013. College of American Pathologists Anatomic Pathology Checklist CAP Accreditation Program. College of American Pathologists; 35–7.Google Scholar
Churukian, CJ 2008. Pigments and minerals. In Bancroft, JD, Gamble, M, eds. Theory and practice of histological techniques. Churchill Livingstone; 233–60.Google Scholar
Economou, M, Schöni, L, Hammer, C, Galván, JA, Mueller, DE, Zlobec, I 2014. Proper paraffin slide storage is crucial for translational research projects involving immunohistochemistry stains. Clin Transl Med. 17;3(1): 4.CrossRefGoogle Scholar
Goldschlager, T, Abdelkader, A, Kerr, J, Boundy, I, Jenkin, G 2010. Undecalcified bone preparation for histology, histomorphometry and fluorochrome analysis. JoVE 35(1707):1–3.Google Scholar
Grizzle, WE, Stockard, CR, Billings, PE 2001. The effects of tissue processing variables other than fixation on histochemical staining and immunohistochemical detection of antigens. J Histotechnol 24:213–19.Google Scholar
Hammond, ME, Hayes, DF, Dowsett, M, Allred, DC, Hagerty, KL, Badve, S, Fitzgibbons, PL, Francis, G, Goldstein, NS, Hayes, M, Hicks, DG, Lester, S, Love, R, Mangu, PB, McShane, L, Miller, K, Osborne, CK, Paik, S, Perlmutter, J, Rhodes, A, Sasano, H, Schwartz, JN, Sweep, FC, Taube, S, Torlakovic, EE, Valenstein, P, Viale, G, Visscher, D, Wheeler, T, Williams, RB, Wittliff, JL, Wolff, AC 2010. American Society of Clinical Oncology/College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. Arch Pathol Lab Med 134(6):907–22.CrossRefGoogle Scholar
Jimenez-Joseph, D, Gangi, MD 1986. Application of Diatex compound in cytology: Use in preparing multiple slides from a single routine smear. Acta Cytol 30(4):446–7.Google ScholarPubMed
Kamino, H, Tam, ST 1991. Immunoperoxidase technique modified by counterstain with azure B as a diagnostic aid in evaluating heavily pigmented melanocytic neoplasms. J Cutan Pathol 18(6):436–9.CrossRefGoogle ScholarPubMed
Kubier, P, Miller, RT 2002. Tissue protection immunohistochemistry: A useful adjunct in the interpretation of prostate biopsy specimens and other selected cases in which immunostains are needed on minute lesions. Am J Clin Pathol 117(2):194–8.CrossRefGoogle ScholarPubMed
Lars, JN, JøRgen, JS, Klaus, G 1991. A method for histological preparation of undecalcified bone sections containing acrylic bone cement. Biotechnic & Histochemistry 66(2):82–6.Google Scholar
Li, X, Deavers, MT, Guo, M, Liu, P, Gong, Y, Albarracin, CT, Middleton, LP, Huo, L 2013. The effect of prolonged cold ischemia time on estrogen receptor immunohistochemistry in breast cancer. Mod Pathol 26(1):71–8.Google Scholar
Li, LX, Crotty, KA, Kril, JJ, Palmer, AA, McCarthy, SW 1999. Method of melanin bleaching in MIB1-Ki67 immunostaining of pigmented lesions: A quantitative evaluation in malignant melanomas. Histochem J 31(4):237–40.CrossRefGoogle ScholarPubMed
Lunn, G, Sansone, EB 1991. The safe disposal of diaminobenzidine. Applied Occupational and Environmental Hygiene 6(1):49–53.Google Scholar
Matthews, JB, Mason, GI 1984. Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue. Histochem J 16(7):771–87.Google Scholar
Medawar, PB 1941. The rate of penetration of fixatives. J Royal Microscopical Society 61:46–7.CrossRefGoogle Scholar
RCPA 2020. Guideline subject: Fixation of Tissues (online). Available at www.rcpa.edu.au/getattachment/d6f7f095-e8b7-45eb-8dcb-6a9d9bd5a88a/Fixation-of-Tissues.aspx# (Accessed 1 November 2020).Google Scholar
Shen, H, Wu, W 2015. Study of melanin bleaching after immunohistochemistry of melanin-containing tissues. Appl Immunohistochem Mol Morphol 23(4):303–7.CrossRefGoogle ScholarPubMed
Torlakovic, EE, Nielsen, S, Vyberg, M, Taylor, CR 2015. Getting controls under control: The time is now for immunohistochemistry. J Clin Pathol 68(11):879–82.CrossRefGoogle ScholarPubMed
Williams, JH, Mepham, BL, Wright, DH 1997. Tissue preparation for immunocytochemistry. J Clin Pathol 50(5):422–8.Google Scholar
Wolff, AC, Hammond, ME, Hicks, DG, Dowsett, M, McShane, LM, Allison, KH, Allred, DC, Bartlett, JM, Bilous, M, Fitzgibbons, P, Hanna, W, Jenkins, RB, Mangu, PB, Paik, S, Perez, EA, Press, MF, Spears, PA, Vance, GH, Viale, G, Hayes, DF 2014. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. Arch Pathol Lab Med 138(2):241–56.CrossRefGoogle Scholar
Yildiz-Aktas, IZ, Dabbs, DJ, Bhargava, R 2012. The effect of cold ischemic time on the immunohistochemical evaluation of estrogen receptor, progesterone receptor, and HER2 expression in invasive breast carcinoma. Mod Pathol 25(8):1098–1105.Google Scholar
Yoo, TK, Moon, HG, Kim, J, Lee, JW, Kim, MK, Lee, E, Kim, J, Han, W, Noh, DY 2014. Effect of prolonged cold ischemic time on immunohistochemical testing of estrogen receptor, progesterone receptor, and HER2 expression in breast cancer. J Clin Onco 32:15 suppl e11505.CrossRefGoogle Scholar

Save book to Kindle

To save this book to your Kindle, first ensure [email protected] is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×