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.
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 (ĸ = 1.00) and moderate/good suitability for the assessment of inflammation (ĸ = 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.
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 tumours in dermatological specimens. Preliminary results found good feasibility when this technique was used to examine prostate cancer (PCa) specimens. Methods We report on the application of FCM in magnet resonance imaging (MRI)-fused prostate biopsies (10 patients, total number of biopsy specimens: n=121) and compare the results to conventional histology. Results Specific structures of the prostatic tissue were very well represented in the FCM images comparable to conventional histology. Prostate carcinoma was diagnosed with good sensitivity (79/68%) and high specificity (100%) by two pathologists with substantial/almost perfect levels of agreement with the results of conventional histology (kappa 0.79/0.86). Depending on the quality of the scans, malignant lesions of 1.8 mm and more in diameter were reliably diagnosed. Smaller lesions were rated as suspect for malignancy, but could not be consistently differentiated from reactive changes. Optimal image qualities were achieved in focus depths of up to 50 µm, whereas deeper scans led to insufficient representation of cytological features. Pre-treatment with acridine orange (AO) did not alter immunoreactivity of the tissue or its feasibility for fluorescence in situ hybridization (FISH) analyses and adequate amounts of DNA could be extracted for further polymerase chain reaction (PCR)-based examinations. Conclusions FCM seems to be a promising tool for the timely diagnosis in cases of PCa in patients requiring therapy. In particular, this technique is a material-sparing method that conserves the biopsies as unfixed material for further analysis such as molecular tumour companion diagnosis.
A previously symptomless 86-year-old man received the first dose of the BNT162b2 mRNA COVID-19 vaccine. He died 4 weeks later from acute renal and respiratory failure. Although he did not present with any COVID-19-specific symptoms, he tested positive for SARS-CoV-2 before he died. Spike protein (S1) antigen-binding showed significant levels for immunoglobulin (Ig) G, while nucleocapsid IgG/IgM was not elicited. Acute bronchopneumonia and tubular failure were assigned as the cause of death at autopsy; however, we did not observe any characteristic morphological features of COVID-19. Postmortem molecular mapping by real-time polymerase chain reaction revealed relevant SARS-CoV-2 cycle threshold values in all organs examined (oropharynx, olfactory mucosa, trachea, lungs, heart, kidney and cerebrum) except for the liver and olfactory bulb. These results might suggest that the first vaccination induces immunogenicity but not sterile immunity.
Postarthroplasty lymphadenopathy should be kept in mind particularly in patients with tumor (eg, melanoma) with a long‐term history of total joint replacement therapy. Microscopy is mandatory in establishing diagnosis and is thus helpful for therapy.