Muskat Linda C, Kerkhoff Yannic, Humbert Pascal, Nattkemper Tim W, Eilenberg Jørgen, Patel Anant V
University of Applied Sciences Bielefeld, Fermentation and Formulation of Biologicals and Chemicals, Faculty of Engineering Sciences and Mathematics, Bielefeld, Germany.
Georg-August-University Goettingen, Agricultural Entomology, Department of Crop Sciences, Goettingen, Germany.
MethodsX. 2021 Jan 7;8:101218. doi: 10.1016/j.mex.2021.101218. eCollection 2021.
The present work describes a new computer-assisted image analysis method for the rapid, simple, objective and reproducible quantification of actively discharged fungal spores which can serve as a manual for laboratories working in this context. The method can be used with conventional laboratory equipment by using bright field microscopes, standard scanners and the open-source software ImageJ. Compared to other conidia quantification methods by computer-assisted image analysis, the presented method bears a higher potential to be applied for large-scale sample quantities. The key to make quantification faster is the calculation of the linear relationship between the gray value and the automatically counted number of conidia that has only to be performed once in the beginning of analysis. Afterwards, the gray value is used as single parameter for quantification. The fast, easy and objective determination of sporulation capacity enables facilitated quality control of fungal formulations designed for biological pest control.•Rapid, simple, objective and reproducible quantification of fungal sporulation suitable for large-scale sample quantities.•Requires conventional laboratory equipment and open-source software without technical or computational expertise.•The number of automatically counted conidia can be correlated with the gray value and after initial calculation of a linear fit, the gray value can be applied as single quantification parameter.
本研究描述了一种新的计算机辅助图像分析方法,用于快速、简单、客观且可重复地定量活性释放的真菌孢子,可为从事该领域工作的实验室提供指南。该方法可通过使用明场显微镜、标准扫描仪和开源软件ImageJ,与传统实验室设备配合使用。与其他通过计算机辅助图像分析进行分生孢子定量的方法相比,本方法在处理大规模样本量方面具有更高的应用潜力。实现更快定量的关键在于计算灰度值与自动计数的分生孢子数量之间的线性关系,该计算只需在分析开始时进行一次。之后,灰度值将作为唯一的定量参数。快速、简便且客观地测定产孢能力有助于对用于生物防治害虫的真菌制剂进行质量控制。
• 适用于大规模样本量的真菌产孢的快速、简单、客观且可重复的定量。
• 需要传统实验室设备和开源软件,无需技术或计算专业知识。
• 自动计数的分生孢子数量可与灰度值相关联,在初步计算线性拟合后,灰度值可作为唯一的定量参数。