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. 2017 May-Aug;21(2):211-217.
doi: 10.4103/jomfp.JOMFP_234_16.

Validation of immunoexpression of tenascin-C in oral precancerous and cancerous tissues using ImageJ analysis with novel immunohistochemistry profiler plugin: An immunohistochemical quantitative analysis

Deepa Rajesh Mane  1 , Alka D Kale  1 , Chetan Belaldavar  1
Affiliations

Validation of immunoexpression of tenascin-C in oral precancerous and cancerous tissues using ImageJ analysis with novel immunohistochemistry profiler plugin: An immunohistochemical quantitative analysis

Deepa Rajesh Mane et al. J Oral Maxillofac Pathol. 2017 May-Aug.

Abstract

Background: Immunohistochemistry (IHC) is a molecular technique that has grown tremendously over the years. However, the assessment is only qualitative which is subjective and causes errors. Due to this limitation, several excellent markers have not gained importance and reached clinical trials. Hence, we aimed to quantify IHC by ImageJ analysis with a novel IHC profiler plugin. ImageJ has not been tried in oral precancerous tissues with minimal attempt for matrix markers.

Aim: This study aimed to validate the quantification of immunoexpression of tenascin-C (TN-C) in oral precancerous tissues and oral squamous cell carcinoma (OSCC) using ImageJ software with IHC profiler plugin.

Materials and methods: After IHC staining for TN-C and image acquisition, ImageJ analysis was performed as per the standard recommended algorithm. Assessment was done by two observers by blinding the histopathological diagnosis. The immunoscore was assessed for interobserver variability using Kohen's kappa statistics.

Results: All our cases were in agreement and found to be statistically significant with P < 0.005. Moderate agreement was for mild dysplasia, moderate dysplasia and oral lichen planus. Substantial agreement was for oral submucous fibrosis and OSCC and almost perfect agreement noted for cases of severe dysplasia.

Conclusion: IHC can now be quantified using freely downloadable software ImageJ analysis in oral precancerous tissues and OSCC. This software with good threshold control can quantify matrix marker such as TN-C. Hence, herewith, we propose that IHC markers should be quantified using ImageJ by our entire oral pathology fraternity so as to have a standard immunoscore for all markers.

Keywords: ImageJ; immunohistochemistry; immunohistochemistry profiler plugin; tenascin-C.

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Conflict of interest statement

There are no conflicts of interest.

Figures

Figure 1
Figure 1
Photomicrograph of the ImageJ software showing immunohistochemistry profiler plugin
Figure 2
Figure 2
Photomicrograph of the ImageJ software showing the selection of “create selection” option
Figure 3
Figure 3
Photomicrograph of the ImageJ software showing the selection of immunohistochemistry profiler macro option
Figure 4
Figure 4
Photomicrograph of immunohistochemistry image of tenascin-C (a), deconvoluted images (b-d), highlighted and selected immunoexpression (e), histogram and log of quantified immunoexpression of tenascin-C in oral epithelial dysplasia (f)
Figure 5
Figure 5
Photomicrograph of immunohistochemistry image of tenascin-C (a), deconvoluted images (b-d), highlighted and selected immunoexpression (e), histogram and log of quantified immunoexpression of tenascin-C in oral lichen planus (f)
Figure 6
Figure 6
Photomicrograph of immunohistochemistry image of tenascin-C (a), deconvoluted images (b-d), highlighted and selected immunoexpression (e), histogram and log of quantified immunoexpression of tenascin-C in oral submucous fibrosis (f)
Figure 7
Figure 7
Photomicrograph of immunohistochemistry image of tenascin-C (a), deconvoluted images (b-d), highlighted and selected immunoexpression (e), histogram and log of quantified immunoexpression of tenascin-C in oral squamous cell carcinoma (f)
See this image and copyright information in PMC

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