It was possible to see structures on an endomicroscopic level detecting different cell structures on a highly focused plane. Additionally, different tissue cause structures, differences between grey and white substances, and arachnoid membranes could be visualized although the scanning field consisted only of 300��m �� 300��m (Figures 2(a)�C2(f)). Figure 2 (a) Grey matter of the pig. (b) White matter of the pig. (c) Arachnoid membrane of the pig. (d) Ventricular wall of the pig. (e) Primary meningioma cell culture. (f) Primary glioblastoma cell culture. The preparation of the tissue before examination proved to be without difficulties. The samples needed no more than a small layer of liquid��in this series isotonic sodium chloride solution��to improve image quality.
Compared to frozen sections done by the neuropathologist, this technique offers a quicker preparation and faster visualisation since staining does not necessarily need to be done. Images of the grey substance, for example, showed a higher density in nuclei compared to white substance, giving impressions of the different structures and scale. The tissue structure was much denser compared to arachnoid mater with a more fibrous pattern including elongated cell bodies and fibrillar cytoplasm. After this first examination, samples were partly stained with methylene blue (MB). For this, the tissue was simply put into MB solution. Depending on the size of the sample, it was best to divide the tissue in small pieces to create a larger contact surface for staining. After 20 minutes of incubation, analysis was performed likewise to the native examination.
Results were mainly not very different from native samples. However, the application of MB helped in the evaluation of the nuclei in selected cases since the nuclei presented themselves darker with a more pronounced contrast depending on how well MB had been absorbed. For further investigation as well as intraoperative use, however, the need of MB staining seems to be questionable, as no significant benefit could be observed. In the second step of analysis, more than 50 tumour specimens were evaluated. It was found that common histological paradigms could not entirely be applied for tumour tissue evaluation. Different endomicroscopic histological criteria were established which were found to be reoccurring amongst different tumour entities.
These criteria mainly included the morphology of the nucleus and its location within the cell, the existence and the shape of the cytoplasm, the presence of psammoma bodies, the cell-to-cell contact, the cellular density Anacetrapib within the specimen, the growth pattern (i.e., diffuse, well sorted), and the presence of blood vessels. Confocal endomicroscopically, glioblastomas showed similarities to normal brain tissue although presenting a higher cellularity. Nuclei were mostly polymorphous and variable in shape, much of how they present themselves in histological findings.