The device will be an outdoor rack enclosure with a power supply connection. ... System (GCS), the concentrator system,
CYTO-WATER
New slet ter # 3 February 2018
EDITORIAL Dear readers, Welcome to the third CYTO-WATER newsletter. After the development of the technologies that constitute the CYTO-WATER device, the different components of the CYTO-WATER system have been successfully validated in t he laboratory and are now being integrated in a single device. Soon, t he validat ion in real environment s w ill t ake place, with the objective of detecting potential issues derived from the installation, operation and maintenance of the device under real conditions. The on-site validation will be carried out first in the Zoo of Barcelona. The validation will continue in Hospital de Terrassa (CST), also in Barcelona. In this third issue, two technologies of the CYTO-WATER device are explained: the microfluidic system , provided by microTEC, and the reader, developed by ICFO. Likewise, the newsletter includes an interview to Helge Bohlman, from microTEC, and Marc Jofre, from ICFO, who offer an overview of the main activities of both partners. Finally, the last section is focused on the CYTO-WATER project Final Workshop, which will be held in Barcelona on May 23rd . The event will be focused on microbiological detection trends and will feature presentations by public administration, end-users and the CYTO-WATER partners, who will present the device. Besides, other European projects related to this issue will be presented. This workshop is mainly intended for end-users, administrations and companies interested in this topic. The CYTO-WATER team keeps working hard to face the last months of the project. For further information about the project, the final event or if you want to subscribe to the newsletter, please feel free to contact us via w w w.cytow ater.eu, where all the project information and related news are periodically updated. Best regards, CYTO-WATER Team
This project has received funding from the European Union?s Horizon 2020 research and innovation programme under Grant Agreement no. 642356.
LATEST NEWS CYTO-WATER is ready for end-users' validat ion CYTO-WATER was designed with the aim of providing a new online system for Legionella and Escherichia coli detection in industrial and environmental waters respectively. The main advantage of this system over traditional microbiology analysis is the reduced time-to-results and the system capability of analysing water samples autonomously. To develop the CYTO-WATER system, t hree technologies are being integrated in a single automated device that can be easily connected direct ly to different facilit ies such as cooling towers or sanitary hot water systems. The system automatically performs the concentration of the required volume, the sample preparation and the detection of the target microorganism. The result s from t he sample concent rat ion are obt ained in around 120 minutes. Besides, the CYTO-WATER system is adapt able for t he detect ion of ot her pat hogens in other sectors, such as food and beverage or clinical, by only modifying the specific markers.
After the development of the technologies that make up the CYTO-WATER device, the different components of the CYTO-WATER system have been successfully validated in t he laboratory by Labaqua to make sure they meet market specifications. After the laboratory validation, the technologies are being integrated into a single plat form.
CYTO-WATER device
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ACHIEVEMENTS So far, the main achievements regarding the system performance are:
The automated concentrator The automated concentration system is capable of processing w ater samples automat ically. The samples (100mL-1L) are concentrated to a final volume of 500?L. This first product from the project, besides being able to be integrated in the CYTO-WATER system, can be used in any type of facility to concent rate a sample automat ically and to be connected w it h ot her detect ion systems. In the laboratory validation stage, the concentrator has been evaluated with 50 natural samples (seawater, cooling towers and hot water sanitary systems) for the subsequent detect ion of Legionella or Escherichia coli.
The microfluidic
Integration
The microfluidic system is t he connect ion bet ween t he concent rator and t he detecto, and it is composed by disposable cart ridges with a microfluidic chamber w here microorganisms labelling and detect ion steps occur. Its performance has been evaluated with different matrices (types of water).
The reader The performance of the reader has been evaluated in three different matrices: pot able and indust rials w aters for Legionella, and seaw ater for Escherichia coli detect ion. More than 100 artificial contaminated samples have been analyzed in parallel by the new reader and other methods (culture and flow cytometry) in order to obtain a correlation. The result s show a correlat ion w it h t he t wo ot her detect ion met hods. Together with this, the system has been tested using natural samples with unknown concentration of microorganisms. The limit of detection of the integrated system will be established in the final laboratory validation.
In parallel to the laboratory validation of these technologies, t he integrat ion of t he system is being carried out . The different components of the CYTO-WATER -concentrator, microfluidic and reader- are being integrated in a single device. The device will be an outdoor rack enclosure with a power supply connection. It will also feature stabilized temperature and dust isolation by a pressurize air fan system. Besides, the device will include the Global Control System (GCS), the concentrator system, a sample preparation fluidic system, the CYTO-WATER reader (isolated from the background light) and one sample chip installed in an automatic controlled rail for sequential measurements.
Somorrostro Beach, Barcelona
The on-site validation will start soon On-site validation of the CYTO-WATER system will be carried out by Cetaqua during the following months. The aim of testing the developed technology in selected demonstration sites is to ident ify potent ial issues derived from t he inst allat ion, operat ion and maintenance under real condit ions. Two demo sites have been selected for CYTO-WATER on-site validation. The first one is Barcelona Zoo, which uses seawater from Somorrost ro Beach as sea animal?s environment. In this demo site, CYTO-WATER analysis will be focused on E.coli, which is the main indicator for bathing water quality. For that purpose, the device will be directly connected to the pipe that continuously provides seawater from the shore to the Zoo.
The second demo site is Hospit al de Terrassa (CST), where the validation will consist in analysing water from the hot sanitary water network. In this case, critical points from the hot water sanitary network will be monitored to detect Legionella.
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INTER VIEWS Helge Bohlmann MSc. in electrical engineering, PhD in precision engineering. Helge Bohlmann met the ?micron? during collaborative work with Volkswagen AG. After different functions joined microTEC in 2000 as R&D project manager focusing today on industrial applications of microfluidics and microsystems. W hat kind of product s does microTEC develop and supply?
W hich are t he most import ant challenges for microTEC in t he fut ure?
microTEC Gesellschaft für Mikrotechnologie mbH (Duisburg, Germany) is active in the development, production and supply of polymer-based 3D printed microstructures and microsystems. For over 20 years, we have offered a full service package to translate product ideas into tangible solutions, from concept and prototyping to volume production. We produce at two sites in Duisburg, Germany and under exclusive license at a partner site in Belgium.
In short, the most important challenge is to stay ahead of competition by listening carefully to the market needs. microTEC is a technology oriented company constantly seeking for new applications of their unique additive manufacturing technologies with still unrivalled performance. However the market for 3D-printing applications is developing quickly and new business strategies and offers to the market must be developed.
In w hat fields does microTEC work? microTEC has been commercializing solutions in those fields where the application of micro technologies plays an important role for the success of a product. In the field of life science, microfluidics plays an important role to realize Lab-on-Chip Systems with applications in environmental monitoring, agriculture and food industry, or medical point-of-care analysis. Furthermore, product solutions are offered, where highly miniaturised systems are needed. This is the case of photonics information and communication applications, where sensors, control systems, batteries, among others, require integration in smallest space. How import ant is innovat ion to microTEC? microTEC is an SME operating in a global and strongly networked environment, where new technologies and their applications quickly find their way out of Europe to all other places in the world. The need for? ideally worldwide - protection of own Intellectual Property (IP) is well known and desired, but expensive and time-consuming. Defending IP is often not possible for SMEs, due to long time frames (until patents are issued) and costs (associated with all procedures). Therefore, a good option for microTEC is to offer innovative product solutions within short development cycles and stay ahead of the competition.
"The main challenge for microTEC is to stay ahead of competition by listening carefully to the market needs" W hat does microTEC expect from t he CYTO-WATER project ? In this project, microTEC will deliver a disposable, custom-made microfluidic cell for the CYTO-WATER System. On top of that, a full microfluidic device is being developed, which is able to perform the CYTO-WATER assay is a seamless way ? from the sample input to the right output for the optical reader-. The expectation from CYTO-WATER project is to be able to deliver new technical solutions and be able to serve the market with a full service package comprising device and other consumables. A new business line will result out of it.
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INTER VIEWS Marc Jofre Marc Jofre received the 2013 PhD Thesis Award for his contribution to industrial research from the Institute of Photonic Sciences (ICFO) in Castelldefels, Spain. He continued his work at ICFO as a postdoctoral researcher. He has a very transversal engineering profile with know-how in different technologies and biology including: optical systems, quantum optics, CMOS image sensors, lens-free microscopy, and immuno-labelling of micro-organisms.
In w hat fields does ICFO focus it s research? Since 2002, ICFO is a centre for research excellence devoted to the science and technology of light. How does ICFO?s research concern t he societ y and t he environment ? ICFO research themes gravitate around solving problems in Health, the Environment, Energy, Information and Safety, all of which are essential to society. ICFO programs create new understanding, solutions, and tools to help industry and the society-at-large cope with its main challenges. How import ant is t he t raining of researchers and technicians to ICFO?
fut ure
ICFO considers graduate students and postdoctoral researchers their most-valued assets. The central mission of the institute is to train the next generation of scientists and technicians by providing a battery of courses, seminars, workshops and mentoring, as well as state-of-the art experimental tools.
"ICFO helps industry and the society-at-large cope with its main challenges" and Technology Transfer (KTT) Office as a key interface with the industrial and corporate worlds. Through its KTT Office, ICFO is particularly proactive in translating newly-generated knowledge into new products and technologies, which has so far led to the constitution of 6 spin-offs since 2002. W hat does ICFO expect from t he CYTO-WATER project ? ICFO expects to learn from the experience of having participated in the design and integration of the CYTO-WATER System. Once the project is completed, ICFO will continue applying the obtained knowledge on the field of bacteria measurement through the creation of a new ICFO spin-off, named SixSenso.
How does research developed w it hin ICFO connect w it h indust ry? ICFO is a leading partner for the optics and photonics industry and integrates a Knowledge www.cytowater.eu
TECHNO LOGIES Microfluidic system A fully automated microfluidic stystem
microTEC has provided an automated microfluidic system for the CYTO-WATER device. The microfluidic system is t he connect ion bet ween t he different component s and serves to process t he concent rated sample. In the core of this system, t he labelling of cells and measuring is performed in a disposable cartridge, which has an internal capacity of 800µl. The CYTO-WATER device will include a four-chamber microfluidic system that does not require human interact ion. As soon as the eluted sample is available in a tube, the system starts processing it automatically and provides a labelled sample at the output, within the targeted time. Besides, the system has an integrated automated cleaning funct ion.
The disposable cartridge The current cartridge, which is manufactured following RMPD® technologies, is based on a polymer st ruct ure, including a top lid and bottom interface made of glass. The shape inside has been optimized to enable bubble-free filling and mixing of the liquids, and the surfaces inside the cartridge do not require any functionalization, which allows low-cost and disposabilit y. A magnetic stirrer is integrated to enable mixing from the outside via an external magnetic field. There are 8 fluidic interfaces to supply the sample, reagents and buffer. It also has a waste channel and ventilation channel. Besides, integrated sealing and a clamping structure ensure a leak-free and ?click-in?attachment to the reader. To manage the supply of the different liquids into the cartridge additional elements are needed, such as external pumps, valves, microelectronic components and software. These addit ional part s supply t he cart ridge w it h t he right amount s of liquids at t he right t ime and for t he proper durat ion.
Cost reduction In contrast to other systems, reagent s and buffers are stored in appropriate cont ainers inside t he device and cooled w here required, which allows reducing the costs of consumables. Precision pumps ensure correct and reliable metering of the volumes and, internally, small volumes reduce reagent consumption. The cartridge works without blister reservoirs for liquids and has no need for cartridge integrated, costly valves. This is a similar approach to modern inkjet printers, where the cartridges are nowadays also kept as simple as possible to reduce the overall costs of disposable parts.
Adaptability
microTEC's disposable cartridge
In conclusion, the automated microfluidic system is designed in a modular fashion. This makes it highly flexible and therefore adapt ive to any kind of assay. There are separate pumps serving each liquid, w hich can be operated independent ly from one ot her. If a different assay needs less reagent the protocol is simply adjusted using the graphical user interface. This makes the system quite versatile for other applications. www.cytowater.eu
TECHNO LOGIES CYTO-WATER's reader by Cedric Hurth
The reader, assembled by ICFO, is the eye of the CYTO-WATER system. It s mission is to see a few tens to a hundred bacteria in a 1mL sample once all the filtering, concentration, and sample preparation steps have been performed. The reader?s detection algorithm can only discriminate between bacteria and debris if the latter are at a low concentration and the bacteria are efficiently labelled providing a high fluorescence signal. The ICFO reader consist s of different element s: - A compact Light-Emit t ing Diode (LED) light source coupled to an optical fibre. - Filters to direct t he opt ical light pat h for excitation and detection with minimal losses. - A detector that quantifies the amount of fluorescent photons received and a soft w are algorit hm that relates it to the number of microorganisms. - Holders and mount s to provide mechanical stability. All these components were selected for their low-cost against performance profile. They are off-the-shelf components with a short lead-time. The main challenges are then to provide stable reliable power sources and acquisition hardware and software to deliver a functional reader.
Integration with the microfluidic system The reader is coupled w it h t he microTEC microfluidic system through the microfluidic chip using collimating optics to ensure efficient signal collection. A bar magnet placed on a motor controls a stir bar integrated in the chip to provide convection during the measurement to ensure an efficient bacteria quantification.
(Top) Connectivity of the fluorescence reader. (Bottom) Assembled optical components.
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UPCOMING EVENTS CYTO-WATER Final Workshop May
rd 23
Barcelona Spain
We are glad to announce that the CYTO-WATER Final Workshop will be held on M ay 23r d in Bar celon a. The event will be focused on microbiological detect ion and will feature presentations by public administ rat ion, end-users and the CYTO-WATER part ners, who will present the device. Finally, other European research project s related to this topic will be presented. This workshop is intended mainly for end-users, administrations and companies interested in this topic. For further information, please visit www.cytowater.eu
The CYTO-WATER project is formed by 6 partners from 4 different European countries, led by Spanish company Labaqua.
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