Ex vivo Fluorescence Confocal Microscopy (FCM) of Perineal Prostate Biopsies (PPB) allows fast-track examination of MRI-guided targeted biopsies in routine diagnostics

Sievert K-D, Hansen T, Titze B, et al. Ex vivo Fluorescence Confocal Microscopy (FCM) of Perineal Prostate Biopsies (PPB) allows fast-track examination of MRI-guided targeted biopsies in routine diagnostics. European Urology. 2023;83(Suppl. 1):S423-S424

Diagnostic Performance of Ex Vivo Fluorescence Confocal Microscopy in the Assessment of Diagnostic Biopsies of the Prostate

Titze U, Hansen T, Brochhausen C, et al. Diagnostic Performance of Ex Vivo Fluorescence Confocal Microscopy in the Assessment of Diagnostic Biopsies of the Prostate. Cancers. 2021;13(22): 5685.Background: Fluorescence confocal microscopy (FCM) is a novel micro-imaging technique providing optical sections of examined tissue. The method has been well established for the diagnosis of tumors in dermatological specimens. Methods: We compare intraoperative diagnoses of the real-time application of FCM in pre-therapeutic prostate biopsies (35 patients, total number of biopsy specimens: n = 438) with the findings of conventional histology. Results: Prostate carcinoma was reliably diagnosed in all patients. Depending on scan quality and experience of the examiner, smaller lesions of well differentiated carcinoma (ISUP1) could not be consistently differentiated from reactive changes. Furthermore, in some cases there was difficulty to distinguish ISUP grade 2 from ISUP grade 1 tumors. ISUP grades 3–5 were reliably detected in FCM. Conclusions: Despite some limitations, FCM seems to be an effective tool for the timely assessment of prostate biopsies enabling reliable diagnosis of prostate cancer in patients requiring therapy

Ex Vivo Fluorescence Confocal Microscopy (FCM) of Prostate Biopsies Rethought: Opportunities of Intraoperative Examinations of MRI-Guided Targeted Biopsies in Routine Diagnostics

Sievert K-D, Hansen T, Titze B, et al. Ex Vivo Fluorescence Confocal Microscopy (FCM) of Prostate Biopsies Rethought: Opportunities of Intraoperative Examinations of MRI-Guided Targeted Biopsies in Routine Diagnostics. Diagnostics. 2022;12(5): 1146.Background: The diagnosis of prostate carcinoma (PCa) requires time- and material-consuming histopathological examinations. Ex vivo fluorescence confocal microscopy (FCM) can detect carcinoma foci in diagnostic biopsies intraoperatively. Methods: MRI-guided and systematic biopsies were identified in a dataset of our previously published study cohort. Detection rates of clinically relevant tumors were determined in both groups. A retrospective blinded trial was performed to determine how many tumors requiring intervention were detectable via FCM analysis of MRI-guided targeted biopsies alone. Results: MRI-guided targeted biopsies revealed tumors more frequently than systematic biopsies. Carcinomas in need of intervention were reliably represented in the MRI-guided biopsies and were identified in intraoperative FCM microscopy. Combined with serum PSA levels and clinical presentation, 91% of the carcinomas in need of intervention were identified. Conclusions: Intraoperative FCM analysis of MRI-guided biopsies is a promising approach for the efficient diagnosis of PCa. The method allows a timely assessment of whether a tumor disease requiring intervention is present and can reduce the psychological stress of the patient in the waiting period of the histological finding. Furthermore, this technique can lead to reduction of the total number of biopsies needed for the diagnosis of PCa

Ex Vivo Fluorescence Confocal Microscopy in Specimens of the Liver: A Proof-of-Concept Study

Titze U, Sievert K-D, Titze B, et al. Ex Vivo Fluorescence Confocal Microscopy in Specimens of the Liver: A Proof-of-Concept Study. Cancers. 2022;14(3): 590.Simple Summary Fluorescence confocal microscopy (FCM) is a novel micro-imaging technique providing optical sections of native tissue. The method is increasingly used for the routine diagnosis of tumors and inflammatory lesions of the skin and shows promising results for the diagnosis of other organ tumors. Very few publications exist about examinations of liver tissue thus far. In this study, we compare findings of FCM-examinations of biopsies and surgical specimens of the liver with the final diagnoses from conventional histology. Ex vivo Fluorescence Confocal Microscopy (FCM) is a technique providing high-resolution images of native tissues. The method is increasingly used in surgical settings in areas of dermatology and urology. Only a few publications exist about examinations of tumors and non-neoplastic lesions of the liver. We report on the application of FCM in biopsies, surgical specimens and autopsy material (33 patients, 39 specimens) of the liver and compare the results to conventional histology. Our preliminary examinations indicated a perfect suitability for tumor diagnosis (kappa = 1.00) and moderate/good suitability for the assessment of inflammation (kappa = 0.4-0.6) with regard to their severity and localization. Macro-vesicular steatosis was reliably detected, micro-vesicular steatosis tended to be underestimated. Cholestasis and eosinophilic granules in granulocytes were not represented in the scans. The tissue was preserved as native material and maintained its quality for downstream histological, immunohistological and molecular examinations. In summary, FCM is a material sparing method that provides rapid feedback to the clinician about the presence of tumor, the degree of inflammation and structural changes. This can lead to faster therapeutic decisions in the management of liver tumors, treatment of hepatitis or in liver transplant medicine

MRI-Guided Targeted and Systematic Prostate Biopsies as Prognostic Indicators for Prostate Cancer Treatment Decisions

Abd Ali F, Sievert K-D, Eisenblätter M, et al. MRI-Guided Targeted and Systematic Prostate Biopsies as Prognostic Indicators for Prostate Cancer Treatment Decisions. Cancers. 2023;15(15): 3915.The standard procedure for the diagnosis of prostate carcinoma involves the collection of 10-12 systematic biopsies (SBx) from both lobes. MRI-guided targeted biopsies (TBx) from suspicious foci increase the detection rates of clinically significant (cs) PCa. We investigated the extent to which the results of the TBx predicted the tumor board treatment decisions. SBx and TBx were acquired from 150 patients. Risk stratifications and recommendations for interventional therapy (prostatectomy and radiotherapy) or active surveillance were established by interdisciplinary tumor boards. We analyzed how often TBx alone were enough to correctly classify the tumors as well as to indicate interventional therapy and how often the findings of SBx were crucial for therapy decisions. A total of 28/39 (72%) favorable risk tumors were detected in TBx, of which 11/26 (42%) very-low-risk tumors were not detected and 8/13 (62%) low-risk tumors were undergraded. A total of 36/44 (82%) intermediate-risk PCa were present in TBx, of which 4 (9%) were underdiagnosed as a favorable risk tumor. A total of 12/13 (92%) high-risk carcinomas were detected and correctly grouped in TBx. The majority of csPCa were identified by the sampling of TBx alone. The tumor size was underestimated in a proportion of ISUP grade 1 tumors. Systematic biopsy sampling is therefore indicated for the next AS follow-up in these cases

Ex Vivo Fluorescence Confocal Microscopy of MRI-Guided Targeted Prostate Biopsies for Rapid Detection of Clinically Significant Carcinomas—A Feasibility Study

Background: MRI-guided prostate biopsies from visible tumor-specific lesions (TBx) can be used to diagnose clinically significant carcinomas (csPCa) requiring treatment more selectively than conventional systematic biopsies (SBx). Ex vivo fluorescence confocal microscopy (FCM) is a novel technique that can be used to examine TBx prior to conventional histologic workup. Methods: TBx from 150 patients were examined with FCM on the day of collection. Preliminary findings were reported within 2 h of collection. The results were statistically compared with the final histology. Results: 27/40 (68%) of the csPCa were already recognized in the intraday FCM in accordance with the results of conventional histology. Even non-significant carcinomas (cisPCa) of the intermediate and high-risk groups (serum prostate-specific antigen (PSA) > 10 or 20 ng/mL) according to conventional risk stratifications were reliably detectable. In contrast, small foci of cisPCa were often not detected or were difficult to distinguish from reactive changes. Conclusion: The rapid reporting of preliminary FCM findings helps to reduce the psychological stress on patients, and can improve the clinical management of csPCa. Additional SBx can be avoided in individual cases, leading to lower rates of complications and scarring in the future surgical area. Additional staging examinations can be arranged without losing time. FCM represents a promising basis for future AI-based diagnostic algorithms

Ex vivo fluorescence confocal microscopy ensures representative tissue in prostate cancer biobanking: A feasibility study

Titze U, Sommerkamp J, Stege C, et al. Ex vivo fluorescence confocal microscopy ensures representative tissue in prostate cancer biobanking: A feasibility study. European Urology. 2023;83(Suppl. 1):S447-S448

Ex Vivo Fluorescence Confocal Microscopy (FCM) Ensures Representative Tissue in Prostate Cancer Biobanking: A Feasibility Study

Titze U, Sommerkamp J, Stege C, et al. Ex Vivo Fluorescence Confocal Microscopy (FCM) Ensures Representative Tissue in Prostate Cancer Biobanking: A Feasibility Study. International Journal of Molecular Sciences. 2022;23(20): 12103.Background: Biobanking of prostate carcinoma is particularly challenging due to the actual cancer within the organ often without clear margins. Frozen sections are to date the only way to examine the biobank material for its tumor content. We used ex vivo fluorescence confocal microscopy (FCM) to analyze biobank samples prior to cryoasservation. Methods: 127 punch biopsies were acquired from prostatectomy-specimens from 40 patients. These biopsies were analyzed with a Vivascope 2500-G4 prior to their transfer to the biobank. In difficult cases, larger samples of the prostatectomy specimens were FCM scanned in order to locate tumor foci. After patient acquisition, all samples were taken from the biobank and analyzed. We compared the results of the FCM examinations with the results of conventional histology and measured the DNA content. Results: With upstream FCM, the tumor content of biobank samples could be determined with high confidence. The detection rate of representative biobank samples was increased due to the rapid feedback. The biobank samples were suitable for further molecular analysis. Conclusion: FCM allows for the first time lossless microscopic analysis of biobank samples prior to their cryoasservation and guarantees representative tumor and normal tissue for further molecular analysis

Circulating dendritic cell precursors in chronic kidney disease: a cross-sectional study

BACKGROUND: Dendritic cells (DC) are professional antigen-presenting cells in the immune system. They patrol the blood as circulating dendritic cell precursors (DCP). Decreased blood DCP count has been shown to be related to atherosclerotic plaque burden. Since chronic kidney disease (CKD) is associated with chronic inflammation and increased cardiovascular risk, the aim of our study was to investigate a potential effect of CKD on circulating DCP numbers especially in patients with a history of cardiovascular disease. METHODS: The number of circulating myeloid (mDCP), plasmacytoid (pDCP), and total DCP (tDCP) was analysed by flow cytometry in 245 patients with CKD stage 3 (with and without known cardiovascular events) and 85 coronary healthy controls. In addition, data were compared with a historical group of 130 patients with known coronary artery disease (CAD). RESULTS: Compared to controls, patients with CKD 3 revealed a significant decrease in circulating mDCP (-29%), pDCP (-43%), and tDCP (-38%) (P < 0.001, respectively). Compared with CAD-patients, the decrease in circulating DCP in CKD was comparable or even more pronounced indicating a potential role for DCP in cardiovascular risk potentiation due to CKD. CONCLUSIONS: Based on previous findings in CAD, the marked decrease of DCP in CKD implicates a potential role for DCP as a mediator of cardiovascular disease. Whether DCP in CKD may act as new cardiovascular biomarkers needs to be established in future prospective trials