Clinical Medical Image Library Clin Med Img Lib 10.23937/2474-3682 2474-3682 ClinMed International Library Wilmington, USA 10.23937 A Human Oocyte Stained for Zona Pellucida Glycoproteins Sousa M 10.23937/2474-3682/1510132 Donor surplus human oocytes were transferred to adhesive slides. Oocytes were fixed with 1% paraformaldehyde in PBS. For permeabilization, cells were incubated in 0.1% Triton X-100 (Sigma) plus 0.1% sodium citrate in PBS. Cells were then incubated with 5% Bovine Serum Albumin (BSA; Sigma) to inhibit non-specific binding. Image ARTICLE 5 2 OPEN ACCESS 10.23937/2474-3682/1510132 A Human Oocyte Stained for Zona Pellucida Glycoproteins Mário Sousa Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, 4050-313 Porto, Portugal Mário Sousa
Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, 4050-313 Porto, Portugal
 21 March 2019 Sousa M 2019 A Human Oocyte Stained for Zona Pellucida Glycoproteins Clin Med Img Lib 10.23937/2474-3682/1510132 2019 Sousa M, et al. © This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Donor surplus human oocytes were transferred to adhesive slides. Oocytes were fixed with 1% paraformaldehyde in PBS. For permeabilization, cells were incubated in 0.1% Triton X-100 (Sigma) plus 0.1% sodium citrate in PBS. Cells were then incubated with 5% Bovine Serum Albumin (BSA; Sigma) to inhibit non-specific binding. Afterwards cells were incubated with a primary monoclonal antibody against human ZP1 (ZP1: sc-365435; Santa Cruz Biotechnology, Santa Cruz, California) for 1 h (37 ºC). For each experiment, a negative control was included. A secondary antibody anti-mouse FITC (sc-516140) conjugated with fluorescein isothiocyanate (FITC; Santa Cruz Biotechnology) was applied to cells for 45 min. Oocytes were not counterstained with DAPI and observed in PBS. Cells were observed in an epifluorescence microscope (Eclipse E400; Nikon, Tokyo, Japan). Experiences were performed in triplicate (Figure 1).

zona pellucida; O: oocyte. Bar: 10 µm. https://clinmedjournals.org/articles/cmil/cmil-5-132-001.jpg