May 27, 2025
Neuromelanin phagocytosis assay
- Lucas Blasco-Agell1,2,
- Meritxell Pons-Espinal1,2,
- Jara Montero-Muñoz1,2,
- Antonella Consiglio1,2,
- Miquel Vila3
- 1Department of Pathology and Experimental Therapeutics, Bellvitge University Hospital- IDIBELL, 08908 Hospitalet de Llobregat, Spain;
- 2Institute of Biomedicine of the University of Barcelona (IBUB), Carrer Baldiri Reixac 15-21, Barcelona 08028, Spain;
- 3VHIR-CIBERNED-ASAP
- Vilalab Public
Protocol Citation: Lucas Blasco-Agell, Meritxell Pons-Espinal, Jara Montero-Muñoz, Antonella Consiglio, Miquel Vila 2025. Neuromelanin phagocytosis assay. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvrq5o3lmk/v1
License: This is an open access protocol 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
Protocol status: Working
We use this protocol and it's working
Created: May 26, 2025
Last Modified: May 27, 2025
Protocol Integer ID: 218934
Keywords: neuromelanin phagocytosi
Funders Acknowledgements:
ASAP
Grant ID: ASAP-020505
Abstract
Neuromelanin phagocytosis assay
Troubleshooting
One-week microglia (hMG) are plated in plastic 12-wp and placed on a Zeiss AXIOIMAGER Z1 maintaining constant levels of temperature (37 ºC) and CO2 (5%).
5 ng/mL of NM are added to cultures and 2 images are taken from every well in duplicate every 2 minutes for 16 hours, using the phase-contrast mode.
Cells are tracked and particles of uptaked NM are counted for every hour.
Cells that died or moved out of the frame are excluded from the analysis.
Accumulated NM is plotted during time and AUC is quantified to perform statistical analysis.
After 24 hours in culture with NM, cell pellets and supernatants of non-stimulated and NM-stimulated cultures are collected and stored at -80 ºC for further analyses.