![]() Here, we briefly describe these controls and how they should be correctly interpreted.Įssential to valid interpretation of immunohistochemical staining is the selection of antibodies that have been validated to detect specific epitopes. Moreover, journals and their editors should require reviewers to comment on whether appropriate minimal positive and negative controls were used. To avoid inaccurate conclusions on a false-positive or a false-negative result, a minimum of a positive and a negative control should be run for all immunohistochemical assays. This sadly influences investigators to have an uncritical attitude towards using valid controls in their studies.Īccordingly, we recommend that the guidelines for authors for all journals should include a requirement that controls must be described and included in manuscripts that report results of immunohistochemical studies. This informal survey sheds light on one reason that controls are often omitted from publications: journals (and, by inference, their reviewers and editors) either do not understand the basic principles of immunohistochemical controls or they do not consider them sufficiently important to require inclusion in their articles. Of the 100 articles in nine journals, up to 80% of papers do not mention controls and 89% of guidelines for authors for these journals do not require or even mention controls. We also reviewed guidelines for authors of all of the selected journals. Specifically, we were interested to know the extent to which control results are readily identifiable in articles that included immunohistochemistry in these journals. To get a snapshot of the “attitude” of many journals for the importance of controls in papers that incorporate immunohistochemical methods, we examined a sample of 29 articles that had immunohistochemical data in the Journal of Histochemistry & Cytochemistry and 71 additional publications in eight other high-impact cell biology and pathology-oriented journals that routinely publish data and images based on immunohistochemistry. This is the best that can be done, but it must be done correctly to be credible and reproducible. With proper controls, however, the investigator can build a convincing case for the presence or absence of a probed molecule. It is important to understand that controls-either positive or negative-can never prove the identity-or the presence or absence-of a molecule in a tissue sample using immunohistochemical techniques. ![]() The possibility of both errors must be considered when designing an immunohistochemical assay. ![]() Both can lead to erroneous scientific conclusions and clinical misdiagnoses. It’s equally important not to conclude that a molecule is present when in fact it is not, as it is to conclude that it is absent when in fact it is present. Simply stated, an immunohistochemical assay that lacks controls cannot be validly interpreted. Furthermore, controls must be described and included in the ensuing publications. Realizing that controls are an essential component of experimental design in all scientific investigations, we insist that valid interpretations of immunohistochemical assays cannot be made in the absence of minimally appropriate controls. Contributing to this problem is the fact that many high-impact journals that publish immunohistochemical data do not require authors to report controls for immunohistochemistry and, what is perhaps more regrettable, the controls that are published often are inappropriate or misinterpreted. This neglect has resulted in the publication of unverified and irreproducible findings in the literature ( Couchman 2014 Collins and Tabak 2014). ![]() 2013), it is the experience of the authors that critical controls are often not performed or reported in ensuing publications. Unfortunately, in the case of immunohistochemistry, although the need for controls is well established ( Baskin 2009 Burry 2000, 2010 2011 Frevert et al. The sine qua non of validating research findings is the use of appropriate controls in the design and performance of experiments and assays.
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