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Fully Automated On-line Flow Cytometry for Microbial Monitoring Life, Death, and Loneliness of Bacteria
Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
RSC | 2018-05-15
Background
2 | 11 Michael Besmer | onCyt Microbiology AG | © 2018
Mai 2013 – September 2016
Monitoring short-term dynamics of bacterial concentrations in natural and engineered aquatic ecosystems
Michael Besmer 2016
Diss. ETH No. 23830
Regionale Wasserversorgung Basel-Landschaft 21
Gesamtsynthese Adrian Auckenthaler und Urs von Gunten (Hrsg.)
Spin-off of Eawag
since February 2017
River water
Spring water
Pumped groundwater
Treated drinking water
Tap water
Besmer et al. (2014) Frontiers in Microbiology
Besmer et al. (2016) Aqua & Gas
Besmer et al. (2016) Nature Scientific Reports
Besmer and Hammes (2016) Water Research
Besmer et al. (2014) Frontiers in Microbiology
Short-term Fluctuations in Bacterial Concentrations are Everywhere
3 | 11 Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
An Established Technology is Moving Forward
Preparation Sample Measurement
Today: manual sampling and sample preparation beofre manual/automated measurement
New possibility: fully automated sampling, sample preparation, and measurement
Sample Preparation and Measurement
4 | 11 Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
Comparability is Important
5 | 11 Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
Take home: Conventional flow cytometry in routine monitoring and targeted monitoring with on-line flow cytometry should be combined/integrated to achieve comprehensive understanding of processes and systems.
Example 1: The Strength of Combined Total and Intact Cell Monitoring
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0
20’000
40’000
60’000
80’000
100’000
350’000
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450’000
500’000
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Inta
ct c
ell c
once
ntra
tion
(cel
ls m
l-1)
Tota
l cel
l con
cent
ratio
n (c
ells
ml-1
)
Take home: The combined measurement of the total and intact cell concentrations provides essential additional information for the assessment of various processes in complex systems – especially with residual disinfectants.
7 days
Michael Besmer | onCyt Microbiology AG | © 2018
Prest and Hammes (2018) Aqua & Gas
RSC | 2018-05-15
Total cell concentration
Intact cell concentration
Example 2: Monitoring Super Low Concentrations after Membranes
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0
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er th
roug
hput
(m3 h
-1)
Tota
l cel
l con
cent
ratio
n (c
ells
ml-1
)
24 hours
Take home: The reliable measurement of super low concentrations makes online flow cytometry suitable for the increasing application of membrane filtration – both for integrity monitoring and operational optimisation. Michael Besmer | onCyt Microbiology AG | © 2018
In preparation
Total cell concentration
Water throughput
RSC | 2018-05-15
0
200
400
600
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1’000
Cell
conc
entr
atio
n (x
103 c
ells
ml-1
) Example 3: Super High Resolution for Rapid Processes and Kinetics
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80 minutes
Tap water Wastewater
Tap water (wash-out)
Shock chlorination
Total cell concentration
Intact cell concentration
Take home: Thanks to super highly resolved measurements, you can assess very rapid processes in great detail and take targeted optimisation steps in an efficient manner if needed. Michael Besmer | onCyt Microbiology AG | © 2018
Besmer et al. (2017) Frontiers in Microbiology
RSC | 2018-05-15
Putting Together the Pieces
9 | 11 Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
Take home: Online measurements are only the starting point of microbiological monitoring. Meaningful interpretation and integration with other information is crucial but necessary experience for facilitation is available.
Summary
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Short-term fluctuations of bacterial concentrations are omnipresent. Hence, it is important to measure them for proper understanding of your systems and processes and to take targeted optimisation where needed. The combined measurement of intact and total cell concentrations often brings the crucial information that makes the difference for managing and ensuring any primary as well as residual disinfection. Online flow cytometry is also a great tool for water treatment steps including membrane filtration, which result in very low concentrations. The analysis and optimisation of particularly rapid processes (e.g., disinfection kinetics) is now possible with novel super-high resolution online flow cytometry. Online flow cytometry supports informed decisions on the design, operation, and optimisation of water treatment and distribution infrastructure. It is a fantastic tool to answer specific questions and might not be needed as a permanent sensors in most places.
Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
Questions and Discussion
11 | 11 Michael Besmer | onCyt Microbiology AG | © 2018 RSC | 2018-05-15
