Abstract
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.
Highlights
Cell viability assays are used in laboratories around the world to determine the effectiveness of drugs and in the exploration of cell death mechanisms
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
A total of 273 dead and 248 alive cells were imaged across the two samples with cell viability confirmed via fluorescence with the absence of any fluorescent signal indicating that the cells are still alive
Summary
Cell viability assays are used in laboratories around the world to determine the effectiveness of drugs and in the exploration of cell death mechanisms. Cellular staining (or labelling) is used to visualise the morphological changes cells undergo during various physiological processes. There is growing concern about the use of stains and dyes and the potential changes that they may induce in cells. Some studies have shown that their introduction alters the native function of cells which can, in turn, lead to incorrect conclusions being drawn from imaging data [1]. The necessity for non-invasive, high contrast techniques that preserve regular functionality is becoming increasingly widely recognised. Such techniques, broadly termed ‘label free imaging’, seek to introduce contrast based on the exploitation of different native photophysical processes without impacting cellular function
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
