TY - JOUR
T1 - Construction and use of a microfluidic dissection platform for long-term imaging of cellular processes in budding yeast
AU - Huberts, Daphne H.E.W.
AU - Sik Lee, Sung
AU - González, Javier
AU - Janssens, Georges E.
AU - Vizcarra, Ima Avalos
AU - Heinemann, Matthias
PY - 2013/6
Y1 - 2013/6
N2 - This protocol describes the production and operation of a microfluidic dissection platform for long-term, high-resolution imaging of budding yeast cells. At the core of this platform is an array of micropads that trap yeast cells in a single focal plane. Newly formed daughter cells are subsequently washed away by a continuous flow of fresh culture medium. In a typical experiment, 50-100 cells can be tracked during their entire replicative lifespan. Apart from aging-related research, the microfluidic platform can also be a valuable tool for other studies requiring the monitoring of single cells over time. Here we provide step-by-step instructions on how to fabricate the silicon wafer mold, how to produce and operate the microfluidic device and how to analyze the obtained data. Production of the microfluidic dissection platform and setting up an aging experiment takes ∼7 h.
AB - This protocol describes the production and operation of a microfluidic dissection platform for long-term, high-resolution imaging of budding yeast cells. At the core of this platform is an array of micropads that trap yeast cells in a single focal plane. Newly formed daughter cells are subsequently washed away by a continuous flow of fresh culture medium. In a typical experiment, 50-100 cells can be tracked during their entire replicative lifespan. Apart from aging-related research, the microfluidic platform can also be a valuable tool for other studies requiring the monitoring of single cells over time. Here we provide step-by-step instructions on how to fabricate the silicon wafer mold, how to produce and operate the microfluidic device and how to analyze the obtained data. Production of the microfluidic dissection platform and setting up an aging experiment takes ∼7 h.
KW - Cell Culture Techniques
KW - Cell Division/physiology
KW - Microfluidic Analytical Techniques/instrumentation
KW - Nanotechnology/methods
KW - Saccharomycetales/physiology
KW - Silicon
UR - http://www.scopus.com/inward/record.url?scp=84879366491&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/nprot.2013.060
DO - https://doi.org/10.1038/nprot.2013.060
M3 - Article
C2 - 23640166
SN - 1754-2189
VL - 8
SP - 1019
EP - 1027
JO - Nature protocols
JF - Nature protocols
IS - 6
ER -