Department of Physiological Diversity, Helmholtz-Centre for Environmental Research (UFZ), Leipzig, Germany.
German Centre for Integrative Biodiversity Research (iDiv) Halle-Jena-Leipzig, Leipzig, Germany.
Cytometry A. 2022 Sep;101(9):782-799. doi: 10.1002/cyto.a.24658. Epub 2022 Jun 7.
Environmental monitoring involves the quantification of microscopic cells and particles such as algae, plant cells, pollen, or fungal spores. Traditional methods using conventional microscopy require expert knowledge, are time-intensive and not well-suited for automated high throughput. Multispectral imaging flow cytometry (MIFC) allows measurement of up to 5000 particles per second from a fluid suspension and can simultaneously capture up to 12 images of every single particle for brightfield and different spectral ranges, with up to 60x magnification. The high throughput of MIFC has high potential for increasing the amount and accuracy of environmental monitoring, such as for plant-pollinator interactions, fossil samples, air, water or food quality that currently rely on manual microscopic methods. Automated recognition of particles and cells is also possible, when MIFC is combined with deep-learning computational techniques. Furthermore, various fluorescence dyes can be used to stain specific parts of the cell to highlight physiological and chemical features including: vitality of pollen or algae, allergen content of individual pollen, surface chemical composition (carbohydrate coating) of cells, DNA- or enzyme-activity staining. Here, we outline the great potential for MIFC in environmental research for a variety of research fields and focal organisms. In addition, we provide best practice recommendations.
环境监测涉及微观细胞和颗粒的定量分析,如藻类、植物细胞、花粉或真菌孢子。传统的使用常规显微镜的方法需要专业知识,耗时且不适合自动化高通量。多光谱成像流式细胞术 (MIFC) 允许从流体悬浮液中每秒测量多达 5000 个颗粒,并且可以同时捕获每个颗粒多达 12 张明场和不同光谱范围的图像,放大倍数高达 60 倍。MIFC 的高通量具有提高环境监测数量和准确性的巨大潜力,例如用于植物传粉者相互作用、化石样本、空气、水或食物质量的监测,这些目前依赖于手动显微镜方法。当 MIFC 与深度学习计算技术结合使用时,还可以实现对颗粒和细胞的自动识别。此外,还可以使用各种荧光染料对细胞的特定部分进行染色,以突出生理和化学特征,包括:花粉或藻类的活力、单个花粉的过敏原含量、细胞的表面化学成分(碳水化合物涂层)、DNA 或酶活性染色。在这里,我们概述了 MIFC 在各种研究领域和焦点生物的环境研究中的巨大潜力。此外,我们还提供了最佳实践建议。