Journal: bioRxiv
Article Title: A surface-engineered microfluidic device for antibody-mediated negative selection of high-quality sperm for assisted reproduction
doi: 10.1101/2025.09.02.673619
Figure Lengend Snippet: Fabrication of gold nano-particle-coated channel slides, containing areas of Anti-PS and progesterone coatings (the ‘NGS Device’) and its effect on immobilising low-quality sperm on the surface. (a) Surface coating with PPOx and addition of GNP to achieve covalent immobilization of COOH-GNP, followed by a second application of PPOx; covalent immobilization of anti-PS antibody (200 µg/mL) onto the glass surface next to the inlet well, and electrostatic adsorption of progesterone onto the half of the glass surface next to the outlet well. (b) Sperm attachment on the channel segment coated with Anti-PS. (c) The surface coated with progesterone exhibited a few or no sperm attachment. (d) Proportion of neat and attached sperm labelled by Annexin v was quantified by fluorescent microscopy. (e) The proportion of sperm labeled with TUNEL was quantified by fluorescent microscopy. (f) Representative fluorescent microscope images of neat samples stained with TUNEL (green) and DAI (blue). (g) Representative fluorescent microscope images of attached sperm on the neat and attached-sperm on Anti-PS-GNP coated surface stained with TUNEL (green) and DAPI (blue). (h, i) Bio-conjugation of Anti-PS using protein G and its effect on the reduction of apoptotic and necrotic sperm populations among recovered sperm through coated channel slides. One-way ANOVA followed by post-hoc t-test was used to evaluate differences between treatment groups. *P ≤ 0.01 and **P ≤ 0.001.
Article Snippet: Progesterone release into the culture medium was quantified using a competitive Progesterone ELISA kit (ALPCO, Cat. No. 11-PROHU-E01), according to the manufacturer’s instructions.
Techniques: Adsorption, Microscopy, Labeling, TUNEL Assay, Staining, Conjugation Assay