Comparison of Real-Time Fluorescence Confocal Digital Microscopy With Hematoxylin-Eosin–Stained Sections of Core-Needle Biopsy Specimens

Abstract

IMPORTANCE Strategies to procure high-quality core-needle biopsy (CNB) specimens are critical for making basic tissue diagnoses and for ancillary testing. OBJECTIVES To investigate acquisition of fluorescence confocal microscopy (FCM) images of interventional radiology (IR)-guided CNB in real time in the radiology suite and to compare the accuracy of FCM diagnoses with those of hematoxylin-eosin (H&E)-stained CNB sections. DESIGN, SETTING, AND PARTICIPANTS In this diagnostic study, FCM imaging of IR-guided CNBs was performed in the radiology suite at a major cancer center for patients with an imaging abnormality from August 1, 2016, to April 30, 2019. The time taken to acquire FCM images and the quality of FCM images based on percentage of interpretable tissue with optimal resolution was recorded. The FCM images were read by 2 pathologists and categorized as nondiagnostic, benign/ atypical, or suspicious/malignant; these diagnoses were compared with those made using H&E-stained tissue sections. Cases with discrepant diagnosis were reassessed by the pathologists together for a consensus diagnosis. Data were analyzed from June 3 to July 19, 2019. INTERVENTIONS Each IR-guided CNB was stained with 0.6mM acridine orange, subjected to FCM imaging, and then processed to generate H&E-stained sections. MAIN OUTCOMES AND MEASURES Mean time taken for acquisition of FCM images, quality of FCM images based on interpretable percentage of the image, and accuracy of diagnostic categorization based on FCM images compared with H&E-stained sections. RESULTS A total of 105 patients (57 male [54.3%]; mean [SD] age, 63 [13] years) underwent IR-guided CNBs in a mean (SD) of 7 (2) minutes each. The FCM images showed at least 20% of the tissue with optimal quality in 101 CNB specimens (96.2%). The FCM images were accurately interpreted by the 2 pathologists in 100 of 105 cases (95.2%) (2 false-positive and 3 false-negative) and 90 of 105 cases (85.7%) (6 false-positive and 9 false-negative). A reassessment of 14 discordant diagnoses resulted in consensus diagnoses that were accurate in 101 of 105 cases (96.2%) (1 falsepositive and 3 false-negative). CONCLUSIONS AND RELEVANCE The ease of acquisition of FCM images of acceptable quality and the high accuracy of the diagnoses suggest that FCM may be useful for rapid evaluation of IR-guided CNBs. This approach warrants further investigation.

Digital Biopsy with Fluorescence Confocal Microscope for Effective Real-time Diagnosis of Prostate Cancer: A Prospective, Comparative Study

“…PB is likely to represent another optimal setting to further evaluate FCM, and we aimed to test the technology into the diagnostic workup of PCa. Similar to other studies [8,9], the ability of FCM to detect prostate neoplastic tissue in biopsy was evaluated in terms of agreement with the conventional HE pathway and an almost perfect agreement was reached (95.1%), suggesting that FCM may be almost as sensitive as HE conventional assessment. The proportion of agreement is high either in the biopsy naïve (97.2%) or in the repeated biopsy (94.2%) setting; further details are reported in Supplementary Table 5.…”

“…The diagnostic capacity of FCM has been proved to be promising in the dermatological field [8] and has also been applied to the interpretation of other human tissues [9].…”

“…Ex vivo FCM has been applied in the field of dermatology [8] and was found to display a high level of agreement with conventional frozen sections analysis for basal cell carcinoma diagnosis [8]. Similarly, when applied to the interpretation of the lung, liver, adrenal gland, kidney, bone, pleura, lymph nodes, and soft tissues, FCM has displayed a 95.2% of agreement with conventional hematoxylin-eosin (HE) [9].…”

Current and future perspectives of digital microscopy with fluorescence confocal microscope for prostate tissue interpretation: a narrative review

“…Ex vivo FCM has been applied in the field of dermatology (3)(4)(5) and has been tested in terms of agreement with conventional frozen sections analysis for the detection of basal cell carcinoma and cutaneous inflammatory diseases, resulting in high levels of accuracy for FCM from a prospective study (3). Similarly, when applied to the interpretation of lung, liver, adrenal gland, kidney, bone, pleura, lymph nodes, and soft tissues, it has displayed a 95.2% of agreement with conventional HE (6).…”

Exploring the utility of Deep Red Anthraquinone 5 for digital staining of ex vivo confocal micrographs of optically sectioned skin

“…Ex vivo confocal microscopy (EVCM), using reflectance and fluorescence contrast, has proven itself to be a useful tool for rapid high‐resolution imaging of frozen–thawed and fresh tissue samples. Acridine orange (AO) is commonly used as a nucleus‐specific fluorescent probe and has become the mainstay in investigations of EVCM of skin as well as in ex vivo surgical pathology in the study of breast, prostate, liver, lung, kidney, and other tissues [1–6]. However, AO binding behavior is not restricted to the nucleus, with several reports describing the labeling of acidic organelles, RNA, matrix proteins, muscle fibers, and hemoglobin [7–10].…”