Anthony Nicholas, Darmanin Connie, Bleackley Mark R, Parisi Kathy, Cadenazzi Guido, Holmes Susannah, Anderson Marilyn A, Nugent Keith A, Abbey Brian
Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Department of Chemistry and Physics, La Trobe Institute for Molecular Science, La Trobe University, Victoria 3086, Australia.
Nanophysics & NIC@IIT, Istituto Italiano Di Tecnologia, Via Enrico Melen 83, 16152 Genoa, Italy.
Biomed Opt Express. 2019 Sep 6;10(10):4964-4974. doi: 10.1364/BOE.10.004964. eCollection 2019 Oct 1.
Characterising and understanding the mechanisms involved in cell death are especially important to combating threats to human health, particularly for the study of antimicrobial peptides and their effectiveness against pathogenic fungi. However, imaging these processes often relies on the use of synthetic molecules which bind to specific cellular targets to produce contrast. Here we study yeast cell death, induced by the anti-fungal peptide, NaD1. By treating yeast as a model organism we aim to understand anti-fungal cell death processes without relying on sample modification. Using a quantitative phase imaging technique, ptychography, we were able to produce label free images of yeast cells during death and use them to investigate the mode of action of NaD1. Using this technique we were able to identify a significant phase shift which provided a clear signature of yeast cell death. Additionally, ptychography identifies cell death much earlier than a comparative fluorescence study, providing new insights into the cellular changes that occur during cell death. The results indicate ptychography has great potential as a means of providing additional information about cellular processes which otherwise may be masked by indirect labelling approaches.
表征和理解细胞死亡所涉及的机制对于应对人类健康威胁尤为重要,特别是在抗菌肽及其对致病真菌有效性的研究方面。然而,对这些过程进行成像通常依赖于使用与特定细胞靶点结合以产生对比度的合成分子。在此,我们研究由抗真菌肽NaD1诱导的酵母细胞死亡。通过将酵母作为模式生物进行研究,我们旨在不依赖样本修饰来理解抗真菌细胞死亡过程。使用定量相成像技术——叠层成像术,我们能够在酵母细胞死亡期间生成无标记图像,并利用这些图像研究NaD1的作用模式。使用该技术,我们能够识别出一个显著的相移,它为酵母细胞死亡提供了清晰的特征。此外,叠层成像术比对比荧光研究更早地识别出细胞死亡,为细胞死亡期间发生的细胞变化提供了新的见解。结果表明,叠层成像术作为一种提供有关细胞过程额外信息的手段具有巨大潜力,否则这些信息可能会被间接标记方法所掩盖。