Asset Details
Do you wish to reserve the book?
Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide
Do you wish to request the book?
Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide
2015
Overview
Background
Viability staining with SYTO9 and propidium iodide (PI) is a frequently used tool in microbiological studies. However, data generated by such routinely used method are often not critically evaluated for their accuracy. In this study we aim to investigate the critical aspects of this staining method using
Staphylococcus aureus
and
Pseudomonas aeruginosa
as the model microorganisms for high throughput studies in microtiter plates. SYTO9 or PI was added alone or consecutively together to cells and the fluorescence intensities were measured using microplate reader and confocal laser scanning microscope.
Results
We found that staining of
S. aureus
cells with SYTO9 alone resulted in equal signal intensity for both live and dead cells, whereas staining of
P. aeruginosa
cells led to 18-fold stronger signal strength for dead cells than for live ones. After counterstaining with PI, the dead
P. aeruginosa
cells still exhibited stronger SYTO9 signal than the live cells. We also observed that SYTO9 signal showed strong bleaching effect and decreased dramatically over time. PI intensity of the culture increased linearly with the increase of dead cell numbers, however, the maximum intensities were rather weak compared to SYTO9 and background values. Thus, slight inaccuracy in measurement of PI signal could have significant effect on the outcome.
Conclusions
When viability staining with SYTO9 and PI is performed, several factors need to be considered such as the bleaching effect of SYTO9, different binding affinity of SYTO9 to live and dead cells and background fluorescence.
Publisher
BioMed Central,BioMed Central Ltd
Subject
