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Investigation of confocal microscopy for differentiation of renal cell carcinoma versus benign tissue. Can an optical biopsy be performed? |
Michael C. Phunga,*(),Andrew R. Rouseb,Jayce Pangilinanc,Robert C. Bellc,Erika R. Bracamontec,Sharfuddeen Mashid,Arthur F. Gmitroe,Benjamin R. Leea
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aDepartment of Urology, University of Arizona College of Medicine, Arizona, USA bDepartment of Medical Imaging, University of Arizona College of Medicine, Arizona, USA cDepartment of Pathology, University of Arizona College of Medicine, Arizona, USA dRinggold Standard Institution, Aminu Kano Teaching Hospital, Kano, Nigeria eBiomedical Engineering, University of Arizona College of Medicine, Arizona, USA |
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Abstract Objective: Novel optical imaging modalities are under development with the goal of obtaining an “optical biopsy” to efficiently provide pathologic details. One such modality is confocal microscopy which allows in situ visualization of cells within a layer of tissue and imaging of cellular-level structures. The goal of this study is to validate the ability of confocal microscopy to quickly and accurately differentiate between normal renal tissue and cancer. Methods: Specimens were obtained from patients who underwent robotic partial nephrectomy for renal mass. Samples of suspected normal and tumor tissue were extracted from the excised portion of the kidney and stained with acridine orange. The stained samples were imaged on a Nikon E600 C1 Confocal Microscope. The samples were then submitted for hematoxylin and eosin processing and read by an expert pathologist to provide a gold-standard diagnosis that can later be compared to the confocal images. Results: This study included 11 patients, 17 tissue samples, and 118 confocal images. Of the 17 tissue samples, 10 had a gold-standard diagnosis of cancer and seven were benign. Of 118 confocal images, 66 had a gold-standard diagnosis of cancer and 52 were benign. Six confocal images were used as a training set to train eight observers. The observers were asked to rate the test images on a six point scale and the results were analyzed using a web based receiver operating characteristic curve calculator. The average accuracy, sensitivity, specificity, and area under the empirical receiver operating characteristic curve for this study were 91%, 98%, 81%, and 0.94 respectively. Conclusion: This preliminary study suggest that confocal microscopy can be used to distinguish cancer from normal tissue with high sensitivity and specificity. The observers in this study were trained quickly and on only six images. We expect even higher performance as observers become more familiar with the confocal images.
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Received: 23 July 2018
Available online: 24 December 2019
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Corresponding Authors:
Michael C. Phung
E-mail: mcphung@gmail.com
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Observer | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | Average | Cases | 118 | 118 | 118 | 118 | 118 | 118 | 118 | 118 | 118 | Correct | 111 | 113 | 113 | 92 | 111 | 106 | 105 | 106 | 107 | Accuracy | 94.1 | 95.8 | 95.8 | 78 | 94.1 | 89.8 | 89 | 89.8 | 90.8 | Sensitivity | 98.5 | 100 | 100 | 100 | 98.5 | 97 | 93.9 | 100 | 98.5 | Specificity | 88.5 | 90.4 | 90.4 | 50 | 88.5 | 80.8 | 82.7 | 76.9 | 81.0 | Positive cases misseda | 1 | 0 | 0 | 0 | 1 | 2 | 4 | 0 | 1 | Negative cases missedb | 6 | 5 | 5 | 26 | 6 | 10 | 9 | 12 | 10 | Area under curve | 0.98 | 0.98 | 0.97 | 0.86 | 0.97 | 0.96 | 0.95 | 0.89 | 0.94 |
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Results of reviewer interpretation of confocal imagery as cancer vs. no cancer.
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Participant number | Sample number | Gold-standard diagnosis | Binary diagnosis | Number of confocal images | 1 | 1 | CCRCC grade 4 | cancer | 6 | 2 | 2 | CCRCC grade 2 | cancer | 9 | 3 | 3 | CCRCC grade 2 | cancer | 6 | 3 | 4 | benign | not cancer | 5 | 4 | 5 | benign | not cancer | 9 | 5 | 6 | CCRCC grade 1 with necrosis | cancer | 6 | 6 | 7 | necrotic tumor | cancer | 5 | 6 | 8 | normal | not cancer | 6 | 7 | 9 | CCRCC grade 1 with fibrosis | cancer | 6 | 7 | 10 | normal with fibrosis | not cancer | 7 | 8 | 11 | CCRCC grade 3 with fibrosis | cancer | 7 | 8 | 12 | normal | not cancer | 9 | 9 | 13 | CCRCC grade 1 | cancer | 7 | 9 | 14 | normal | not cancer | 9 | 10 | 15 | CCRCC grade 1 with hemosiderin | cancer | 8 | 11 | 16 | CCRCC grade 1 | cancer | 6 | 11 | 17 | normal | not cancer | 7 |
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Overview of pathology specimens obtained from enrolled subjectsa.
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Example confocal images (20×) of normal kidney and kidney cancer. The normal kidney (A) has clearly identifiable normal renal tubules. (B) Renal cell carcinoma in which there is a higher cellular density, and when magnified, the shape of the nuclei is also more heterogeneous.
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Operating characteristic curves from eight observers.
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