Porcine eyes were obtained from the ex vivo perfusion experiments. For TEM analysis, cornea-scleral slices from one quadrant per eye were fixed in Karnovsky’s solution (2.5% glutaraldehyde and 2.5% paraformaldehyde in a 0.1 M cacodylate buffer) for 24 h [23]. After rinsing in the 0.1 M cacodylate buffer, postfixation was accomplished in a mixture of 1% OsO4 and 0.8% potassium ferrocyanide in a 0.1 M cacodylate buffer for 3.5 h at 48 °C. The eyes were then dehydrated in a graded series of ethanol and embedded in Epon (Serva, Heidelberg, Germany). Semithin sections (1 µm) were collected on uncoated glass slides and stained with methylene blue/azure II (LIT). Ultrathin sections were mounted on uncoated copper grids, stained with uranyl acetate and lead citrate, and examined on a Zeiss Libra transmission electron microscope (Carl Zeiss AG). The number of NPs in the trabecular meshwork was quantified and related to an area of 1000 μm². Particles were counted in the entire trabecular meshwork and divided into the outer and inner part of the trabecular meshwork. Particles that did not penetrate deeply into the trabecular meshwork were attributed to the outer trabecular meshwork, while particles that were found deeper in the trabecular meshwork were assigned to the inner trabecular meshwork. Images were analyzed with ImageSP (TRS, Moorenweis, Germany) and QuPath (version 0.2.3, University of Edinburgh, Edinburgh, UK).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.