Aknoun Sherazade, Savatier Julien, Bon Pierre, Galland Frédéric, Abdeladim Lamiae, Wattellier Benoit, Monneret Serge
Aix-Marseille Université, Centre National de la Recherche Scientifique, Centrale Marseille, Institut Fresnel UMR 7249, 13013 Marseille, FrancebPHASICS S.A., Parc technologique de Saint Aubin, Route de l'Orme des Merisiers, 91190 Saint Aubin, France.
Aix-Marseille Université, Centre National de la Recherche Scientifique, Centrale Marseille, Institut Fresnel UMR 7249, 13013 Marseille, France.
J Biomed Opt. 2015;20(12):126009. doi: 10.1117/1.JBO.20.12.126009.
Single-cell dry mass measurement is used in biology to follow cell cycle, to address effects of drugs, or to investigate cell metabolism. Quantitative phase imaging technique with quadriwave lateral shearing interferometry (QWLSI) allows measuring cell dry mass. The technique is very simple to set up, as it is integrated in a camera-like instrument. It simply plugs onto a standard microscope and uses a white light illumination source. Its working principle is first explained, from image acquisition to automated segmentation algorithm and dry mass quantification. Metrology of the whole process, including its sensitivity, repeatability, reliability, sources of error, over different kinds of samples and under different experimental conditions, is developed. We show that there is no influence of magnification or spatial light coherence on dry mass measurement; effect of defocus is more critical but can be calibrated. As a consequence, QWLSI is a well-suited technique for fast, simple, and reliable cell dry mass study, especially for live cells.
单细胞干质量测量在生物学中用于跟踪细胞周期、研究药物作用或探究细胞代谢。采用四波横向剪切干涉术(QWLSI)的定量相成像技术可实现细胞干质量的测量。该技术的设置非常简单,因为它集成在类似相机的仪器中。它只需连接到标准显微镜上,并使用白光照明源。首先解释其工作原理,从图像采集到自动分割算法以及干质量定量。针对不同类型的样本和不同实验条件,对整个过程的计量学进行了研究,包括其灵敏度、重复性、可靠性和误差来源。我们表明,放大倍数或空间光相干性对干质量测量没有影响;散焦的影响更为关键,但可以进行校准。因此,QWLSI是一种非常适合用于快速、简单且可靠的细胞干质量研究的技术,特别是对于活细胞。