Winters Michela, Aru Violetta, Howell Kate, Arneborg Nils
School of Agriculture & Food, Faculty of Veterinary & Agricultural Science, University of Melbourne, Parkville, 3010, Australia.
Department of Food Science, University of Copenhagen, Frederiksberg, 1958, Denmark.
Biotechniques. 2022 Mar;72(3):100-103. doi: 10.2144/btn-2021-0120. Epub 2022 Feb 7.
Filamentous growth in is a stress response commonly induced under nutrient deprivation and by certain alcohols. It is a compound phenotype characterized by pseudohyphal growth, invasion and a shift to more polarized budding. Previous methods have not allowed the time-resolved determination of filamentous growth. Here we present a new method for budding pattern characterization that enables the measurement of filamentous growth and metabolite concentration during yeast cell growth at precise time intervals. By combining chemical cell immobilization and single-cell imaging using an oCelloScope™, this method provides more accurate budding pattern classification compared with previous methods. The applications of the method include, for example, investigation of quorum sensing-controlled yeast filamentous growth and metabolism under stress and identification of toxic metabolites.
丝状生长是一种应激反应，通常在营养剥夺和某些醇类作用下诱导产生。它是一种复合表型，其特征为假菌丝生长、侵袭以及向更极化的出芽转变。以往的方法无法实现对丝状生长的时间分辨测定。在此，我们提出一种用于出芽模式表征的新方法，该方法能够在酵母细胞生长过程中以精确的时间间隔测量丝状生长和代谢物浓度。通过结合化学细胞固定和使用oCelloScope™进行单细胞成像，与以往方法相比，该方法能提供更准确的出芽模式分类。该方法的应用包括，例如，研究群体感应控制的酵母在应激条件下的丝状生长和代谢，以及鉴定有毒代谢物。
