Investigation of daughter cell dissection coincidence of single budding yeast cells immobilized in microfluidic traps
| Autor: | Feng Xu, Dejing Pan, Yingying Wang, Zhen Zhu, Yangye Geng, Zixin Wang, Mario Andrea Marchisio, Xingyu Xu |
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| Rok vydání: | 2020 |
| Předmět: |
Materials science
Cell division Saccharomyces cerevisiae Microfluidics Finite Element Analysis 02 engineering and technology Dissection (medical) 01 natural sciences Biochemistry Analytical Chemistry Single-cell analysis medicine Budding biology 010401 analytical chemistry Equipment Design Cells Immobilized Microfluidic Analytical Techniques 021001 nanoscience & nanotechnology biology.organism_classification medicine.disease Budding yeast Yeast 0104 chemical sciences Saccharomycetales Biophysics Hydrodynamics Single-Cell Analysis 0210 nano-technology Cell Division |
| Zdroj: | Analytical and bioanalytical chemistry. 413(8) |
| ISSN: | 1618-2650 |
| Popis: | Microfluidic methodologies allow for automatic and high-throughput replicative lifespan (RLS) determination of single budding yeast cells. However, the resulted RLS is highly impacted by the robustness of experimental conditions, especially the microfluidic yeast-trapping structures, which are designed for cell retention, growth, budding, and daughter cell dissection. In this work, four microfluidic yeast-trapping structures, which were commonly used to immobilize mother cells and remove daughter cells for entire lifespan of budding yeast, were systematically investigated by means of finite element modeling (FEM). The results from this analysis led us to propose an optimized design, the yeast rotation (YRot) trap, which is a “leaky bowl”–shaped structure composed of two mirrored microcolumns facing each other. The YRot trap enables stable retention of mother cells in its “bowl” and hydrodynamic rotation of buds into its “leaky orifice” such that matured progenies can be dissected in a coincident direction. We validated the functions of the YRot trap in terms of cell rotation and daughter dissection by both FEM simulations and experiments. With the integration of denser YRot traps in microchannels, the microfluidic platform with stable single-yeast immobilization, long-term cell culturing, and coincident daughter dissection could potentially improve the robustness of experimental conditions for precise RLS determination in yeast aging studies. |
| Databáze: | OpenAIRE |
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