title: Using dielectrophoresis to study the dynamic response of single budding yeast cells to Lyticase.
abstract: Budding yeast cells are quick and easy to grow and represent a versatile model of eukaryotic cells for a variety of cellular studies, largely because their genome has been widely studied and links can be drawn with higher eukaryotes. Therefore, the efficient separation, immobilization, and conversion of budding yeasts into spheroplast or protoplast can provide valuable insight for many fundamentals investigations in cell biology at a single cell level. Dielectrophoresis, the induced motion of particles in non-uniform electric fields, possesses a great versatility for manipulation of cells in microfluidic platforms. Despite this, dielectrophoresis has been largely utilized for studying of non-budding yeast cells and has rarely been used for manipulation of budding cells. Here, we utilize dielectrophoresis for studying the dynamic response of budding cells to different concentrations of Lyticase. This involves separation of the budding yeasts from a background of non-budding cells and their subsequent immobilization onto the microelectrodes at desired densities down to single cell level. The immobilized yeasts are then stimulated with Lyticase to remove the cell wall and convert them into spheroplasts, in a highly dynamic process that depends on the concentration of Lyticase. We also introduce a novel method for immobilization of the cell organelles released from the lysed cells by patterning multi-walled carbon nanotubes (MWCNTs) between the microelectrodes.
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, VIC, 3001, Australia, shiyang.tang@rmit.edu.au.
authors.forename: Shi-Yang
authors.initials: SY
authors.lastname: Tang
authors.forename: Pyshar
authors.initials: P
authors.lastname: Yi
authors.forename: Rebecca
authors.initials: R
authors.lastname: Soffe
authors.forename: Sofia
authors.initials: S
authors.lastname: Nahavandi
authors.forename: Ravi
authors.initials: R
authors.lastname: Shukla
authors.forename: Khashayar
authors.initials: K
authors.lastname: Khoshmanesh
mesh_terms: D018914:Cells, Immobilized / Q000737:chemistry / Q000166:cytology; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry / Q000494:pharmacology*; D008839:Microelectrodes; D009097:Multienzyme Complexes / Q000737:chemistry / Q000494:pharmacology*; D037742:Nanotubes, Carbon / Q000737:chemistry; D010447:Peptide Hydrolases / Q000737:chemistry / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000187:drug effects; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D013104:Spheroplasts
chemical_list: D009097:Multienzyme Complexes; D037742:Nanotubes, Carbon; C025310:lyticase; D004693:Glucan Endo-1,3-beta-D-Glucosidase; D010447:Peptide Hydrolases