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Introduction: Thick histological samples are difficult to image without proper tissue clearing methods. Among these methods ethyl cinnamate (ECi)-based clearing preserves antigenicity and is compatible with immunofluorescent labeling. In contrast to many other clearing protocols, ECi-based clearing is fast and is done as a final step after standard immunofluorescent labeling protocols. Aim: We aimed to develop a simple method of ECi-based tissue clearing approach of thick (100 µm) sections attached to a slide glass. Materials and methods: Samples of human colorectal cancer underwent fixation and frozen section. We used indirect immunofluorescence to label Von-Willebrand factor, which is expressed in blood vessels. After completion of labeling and nuclear DAPI staining, the material was dehydrated using alcohols. Finally, the material was cleared and mounted with ECi and subsequently visualized via standard widefield epifluorescence microscopy. Image analysis of z-stacks were evaluated for the depth of visible signals and compared them to non-cleared samples. Results: Compared to the non-cleared sections, the ECi-cleared sections provided a 2.5-fold increase in the observable tissue thickness following immunofluorescent staining. Further, the proposed approach is time efficient (4 days from tissue preparation to final imaging) as compared to other tissue clearing methods and low cost as it minimizes the use of large amounts of reagents. Conclusion: ECi-based clearing is a very effective simple extension of standard immunofluorescent protocols and can be implemented in future experiments.

Low grade fibromyxoid sarcoma (LGFMS) is an uncommon variant of fibrosarcoma with high risk of local recurrence, immense metastatic potential and frequently protracted period between tumour presentation and metastasis. This unusual malignancy rarely affects the region of the head and neck which makes cases of laryngeal LGFMS extremely infrequent. To date, LGFMS of the larynx has been scatteredly mentioned in the literature. Neither incidence nor causes and risk factors for laryngeal LGFMS have been clarified so far. To the authors’ knowledge, this is the first case report that discusses the clinical course, imaging diagnosis, histopathological evaluation and surgical approach to radiation-induced laryngeal LGFMS.We present a case of a 70-year-old man who developed a LGFMS after previous radiotherapy (RT) for squamous cell carcinoma (SCC) of the larynx. The latency period between the time of radiation exposure and the diagnosis of LGFMS was twenty-seven months. After re-confirming the diagnosis with second biopsy and extensive imaging evaluation the patient was subjected to an open partial resection of the larynx. Owing to the rarity of the tumour, there is no established protocol with follow-up recommendations.This case highlights the importance of considering the RT history of the patient in order to monitor radiotherapy-related complications, including the occurrence of LGFMS.