Reliable replicative lifespan determination of yeast with a single-channel microfluidic chip
Saccharomyces cerevisiae is a powerful model for aging research due to its short lifespan and genetic malleability. Microfluidic devices offer an attractive approach enabling rapid monitoring of hundreds of cells during their entire replicative lifespan (RLS). Yet, key operational issues such as contaminations, cell loss, and cell-aggregates-dependent flow obstruction can hinder RLS experiments. We report the development of a microfluidic device configuration that effectively prevents flow blockage. We conducted comprehensive performance characterization, evaluating trapping efficiency, cell retention, budding orientation, and cell aggregate formation. The optimized device successfully supported long-term culturing and reliable RLS measurements of budding yeast strains. For accurate lifespan determination, a detailed workflow is provided that includes device fabrication, live microscopy setup, and characterization of cell age distribution. This work describes an accessible and reliable microfluidic device for yeast RLS studies, promoting further exploration in aging research.
Authors
- Jerónimo Bravo (ORCID: https://orcid.org/0000-0001-6695-2846)
- Guido Berlin
- Pablo S. Aguilar (ORCID: https://orcid.org/0000-0002-9312-2474)
- Valentina Salzman (ORCID: https://orcid.org/0000-0002-2451-7632)
- Marjolaine Willems (ORCID: https://orcid.org/0000-0002-2959-0935)
- Laura C. Estrada (ORCID: https://orcid.org/0000-0003-4024-4627)
- Marı́a E. Torres (ORCID: https://orcid.org/0000-0002-1512-9507)
- Martín G. Bellino (ORCID: https://orcid.org/0000-0002-6713-6688)
- Nahuel Tarkowski
- Francisco G. Correa Tedesco
- M. Mercuri (ORCID: https://orcid.org/0000-0002-3000-8017)
- Dante G. Mercado
Institutions
- Consejo Nacional de Investigaciones Científicas y Técnicas (AR)
- National University of General San Martín (AR)
- Universidad de Buenos Aires (AR)
- Institut Català de Nanociència i Nanotecnologia (ES)
- Universidad Nacional de San Martín (PE)
- Instituto de Física de Buenos Aires (AR)
- Fundación Ciencias Exactas y Naturales (AR)
- Comisión Nacional de Energía Atómica (AR)
Publication Details
- Journal
- Biology Open
- Published
- 2024-10-31
- DOI
- https://doi.org/10.1242/bio.060596
- Primary Topic
- Gene Regulatory Network Analysis
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Consejo Nacional de Investigaciones Científicas y Técnicas
- Universidad de Buenos Aires
- Agencia Nacional de Promoción Científica y Tecnológica
- Secretaría de Ciencia y Técnica, Universidad de Buenos Aires