Determination of Single Cell Growth Kinetics with Microcantilever Sensors

RAN Pub Date : 2016-04-01 DOI:10.11159/ICNNFC16.125
A. Wańczyk, Bogdan Łabędź, Z. Rajfur
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引用次数: 0

Abstract

Extended Abstract Study of different physical and biochemical parameters of a cell during its life cycle and analysis of underlying regulatory processes are crucial in biomedical and life sciences [1, 2]. One of such parameters is a mass of the cell which is inherently associated with important cellular processes like growth or reproduction. Despite decades of research on cell cycle, there is still a great need for better understanding of many involved process on a single cell level. Such measurements are aided by nano and micro electromechanical systems (NEMS/MEMS) to which nanoand microcantilever biosensors belong to. Cantilever-based devices convert biological or chemical interactions into mechanical response such as cantilever bending amplitude or its resonance frequency change. Objective of this study was the establishment of experimental procedures and analytical tools for comprehensive studies of single cell life cycle using microcantilever biosensors. This work investigated growth kinetics of single cells of Saccharomyces cerevisiae (Instaferm, Lallemand, Poland). S. cerevisiae are commonly used in the cell studies as a model eukaryotic cell. They are widely available and easy to maintain. Here, we demonstrate that it is possible to measure mass changes of single cell during the cell budding using microcantilever biosensor. Analysis of mass changes was based on measuring cantilever resonance frequency changes due to cells adhesion and reproduction. Another important factor in this analysis was the determination of individual cell positions along the cantilever length [3]. Their positions were identified from respective microscopy images. The experiments were performed with Cantisens CSR-801 Concentris (Zurich, Switzerland) cantilever biosensor and Axio Observer Z1 Zeiss (Jena, Germany) microscope. The results of our study provide basic groundwork for further studies of single cell cycle parameters. This work also shows that combination of microcantilever-based sensors and microscopy techniques can be a powerful tool in cell mass kinetics analysis at a single cell level.
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微悬臂传感器测定单细胞生长动力学
在生物医学和生命科学中,研究细胞生命周期中不同的物理和生化参数以及分析其潜在的调控过程是至关重要的[1,2]。其中一个参数是细胞的质量,它与重要的细胞过程(如生长或繁殖)有内在的联系。尽管对细胞周期的研究已经进行了几十年,但在单细胞水平上对许多相关过程的理解仍有很大的需要。这种测量是由纳米和微悬臂生物传感器所属的纳米和微机电系统(NEMS/MEMS)辅助的。基于悬臂梁的装置将生物或化学相互作用转化为机械响应,如悬臂梁弯曲幅度或其共振频率变化。本研究的目的是建立利用微悬臂生物传感器对单细胞生命周期进行综合研究的实验程序和分析工具。本研究研究了酿酒酵母(Instaferm, Lallemand,波兰)单细胞的生长动力学。酿酒葡萄球菌在细胞研究中经常被用作真核细胞的模型。它们广泛可用且易于维护。在这里,我们证明了用微悬臂生物传感器测量细胞出芽过程中单细胞的质量变化是可能的。质量变化的分析是基于测量由于细胞粘附和繁殖引起的悬臂共振频率变化。该分析的另一个重要因素是确定单个细胞沿悬臂长度的位置[3]。从各自的显微镜图像中确定了它们的位置。实验采用瑞士苏黎世悬臂式生物传感器Cantisens CSR-801、德国耶拿Axio Observer Z1 Zeiss显微镜进行。本研究结果为进一步研究单细胞周期参数提供了基础。这项工作还表明,基于微悬臂的传感器和显微镜技术的结合可以成为单细胞水平上细胞质量动力学分析的有力工具。
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