Basic scienceConfocal laser scanning microscopy of urinary bladder after intravesical instillation of a fluorescent dye
Section snippets
Material and methods
The study was performed on the bladder of male Copenhagen rats. Experimental studies were done using a CLSM (Zeiss LSM 410 invert beam scanning) and a 40× water immersion objective (Zeiss, C-Apochromat, W Korr) with a numeric aperture of 1.2. For CFM, beam splitters FT 515 and FT 655 (Zeiss), as well as filters LP 515 and RG 665 for excitation at 488 nm (argon laser) and 633 nm (helium-neon laser), were used, respectively. The bladder was illuminated with a power of 38 μW at 488 nm and 45 μW at
Results
CFM allowed imaging of cellular details of the entire urothelium. This included the large, flat, umbrella cells on the surface, as well as the underlying intermediate and basal urothelial cells. In addition, below the urothelium, a tight network of capillaries and the underlying superficial layers of the lamina propria could be studied (Fig. 1). The CFM images were close to those obtained by standard microscopy after conventional hematoxylin-eosin staining (Fig. 1). Cell structure, such as
Comment
CFM enables the study of the microscopic anatomy of normal bladder mucosa and superficial layers of the lamina propria. The study was performed on intact tissue, representing the in vivo situation. To our knowledge, this is the first time that rat bladder tissue has been studied by CFM using a fluorescent marker like SYTO 17. Regarding the possible applications in humans, the use of SYTO 17 is purely experimental. The potential toxicity and mutagenicity, as well as the optimal incubation
Conclusions
To date, the current CFM system allows the study of horizontal tissue planes only. This is a disadvantage, especially if one looks at carcinoma in situ, for which it is important to assess the maturation of the urothelium. However, through automation of the diagnostic process and the use of computer-based technology during endoscopy, cancer screening may become three-dimensional, less time consuming, less invasive, and highly efficient at a lowered expense.7 In the future, with the development
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Intraoperative Tissue Characterization and Imaging
2009, Urologic Clinics of North AmericaCitation Excerpt :CFM can also provide enough resolution to allow distinction between some differentiated and undifferentiated cancers.15 Koenig and colleagues13 have demonstrated the use of CFM for endoscopic microscopy of the bladder and opened the possibility of using this technology for real-time in vivo assessment of bladder lesions. CFM allowed imaging of cellular details of the entire urothelium.
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