The lab-free CBRN detection device for the identification of biological pathogens on nucleic acid and immunological level as lab-on-a-chip system applying multisensor technologies

Final Report Summary - MULTISENSE CHIP (The lab-free CBRN detection device for the identification of biological pathogens on nucleic acid and immunological level as lab-on-a-chip system applying multisensor technologies)

Executive Summary:
The goal of the “Multisense Chip” project was to build an integrated “sample-in-result-out” analytical system for the detection and identification of biological pathogens. Two user scenarios were covered within the project resulting in two different systems: A “one-time measurement system” enabling the analysis of samples on demand and a “permanent monitoring system” ensuring a permanent monitoring of potential contamination of sensitive areas with biological pathogens. The “Multisense Chip” systems with a lab-on-a-chip as core component allows for a detection of biological pathogens on molecular and immunological level.
Main characteristics of the “Multisense Chip Analyzer” are a “sample-in-results-out” operation manner, an extremely easy handling even suited for untrained personnel, and finally a fast response time of roughly 60 minutes for the overall process, and finally a lab-free approach. The “Multisense Chip Analyzer” can be operated at the place of need, what differs from the current state of the art, were all biologically challenging steps require equipped laboratories and trained staff.
At first, a set of typical B-agents for e.g. terroristic attacks was chosen as target species that the “Multisense Chip Analyzer” needs to detect comprising Yersinia pestis, Francisella tularensis, Burkholderia mallei, Burkholderia pseidomallei, Brucella melitensis, Brucella abortis, Coxiella burnetti and Bacillus anthracis.
To cope with the envisaged “sample-in-result-out” operation procedure, the “Multisense Chip Analyzer” includes the complete process chain from sample collection followed by sample transfer into the lab-on-a-chip system. The subsequent processes on chip encompass sample preparation for direct extraction of target molecules of various kinds of matrices, the biochemical detection method, and, finally, the readout via different sensor technologies.
Keys to reaching the above project goals were advanced sensor technologies, lab-on-a-chip concepts with its intrinsic advantages of miniaturization and innovative instrumentation. Several enabling technologies based on micro- and nanotechnologies came into place for the realization of the various components of the lab-on-a-chip system, biochemical processes, as well as scientific instrumentation. Information & communication technologies coping with the control of the system, the analysis and presentation of the analytical results, as well as with the communication of eventual findings to first responders and respective authorities were involved.
Due to the multifaceted tasks resulting from bio-analytical, microfluidic, engineer-technical, communication-technical, and, finally, end-user based requirements, the “Multisense Chip” project was composed by a high multidisciplinary project team. Taking the competencies of the “Multisense Chip” project partners in mind, it was ensured that their complementary expertise and overlapping know-how in the critical project paths not only covered the above demand of the project, but directly implied a contingency plan to guarantee a smooth course of the project.

Project Context and Objectives:
The terroristic attacks with non conventional weapons as the B-agents attacks in the US of October 2001, or the avian flu in Asia and Europe, or the Ebola virus outbreaks in West Africa demonstrate not only the limited capacities of the different national analytical labs, but also highlights the handling of such major threats and the proper response to them being massively hindered by the lack of rapid, safe and portable detection and identification methods to securely determine an infection or contamination of people, animals, food or sensitive infrastructure.
Most of the presently available commercial detection methods for biological pathogens are based on three fundamental methods and have different drawbacks. Microbiological and immunological procedures are usually quite cost-efficient, but can only be carried out within a suitably equipped laboratory infrastructure. Furthermore, they are time-consuming and not very sensitive. Analytical instruments based on nucleic acid detection are very specific with respect to the different biological pathogens and also very sensitive, but usually they cannot be used in a mobile environment, are time-consuming and complex, and can only be handled by trained staff. Sample preparation and analysis still need at least several hours, which is unacceptable in critical situations like those involving victims of B-agents and critical infrastructure.
The potential risk of airborne pathogens illustrates the urgent need for sensitivity: airborne pathogenic bacteria require sample enrichment from a large volume of air, its transfer into a small volume of liquid, followed by a detection method as sensitive as possible, ideally reaching a detection limit of one DNA copy per assay. Highly infectious bacteria like Francisella tularensis, Brucella melitensis or Coxiella burnetii have a low infectious dose. Therefore, any device assumed to be used in continuous environmental monitoring should be able to detect minute amounts of hazardous microorganisms in air samples. To ensure early warning, highly efficient air sampling and concentration of airborne particles are critical steps in continuous surveillance.
The final goal of the Multisenses Chip project was the development of analytical instruments to carry out with confidence the detection and identification of biological pathogens on the molecular and immunological levels.
The systems are based on a control instrument and a lab-on-a-chip as a consumable. They do not only carry out the detection reaction, namely the PCR and immunoassays, but also the sample enrichment and the extraction of the target molecules. Finally, the systems also carry out the detection via electrochemical and optical means. All functions after sample uptake, concentration and transfer are integrated on-chip to avoid all manual handling steps as well as the need for equipped laboratories and trained personnel. This means a “sample in, result out” system is at hand, directly highlighting the analytical results on the molecular and immunological levels.
The systems developed in the Multisense Chip project cover two user scenarios:
1. Permanent monitoring of airborne pathogens: In this user scenario bacterial pathogens are spread into air, meaning that large volumes of air have to be sampled, target molecules have to be transferred in a liquid phase that is concentrated and a defined volume is moved on the lab-on-a-chip device.
2. One-time measurement of liquid and swab samples: This scenario implies that bacteria or other pathogens are exposed on surfaces like letters inside envelopes, dispersed onto food or in liquids/beverages and that the suspicious sample will be collected with a swab or directly as liquid. This required the establishment of a methodology to collect the sample in a standard manner, to extract the sample from the swab, and to quantitatively transfer the sample from the sampling vessel to the chip.
To cover both user scenarios “permanent monitoring” and “one-time-use” different lab-on-a-chip solutions and two control instruments to run the chip were developed.
The overall project target, the realization of a “sample in, result out” system for the detection and identification of biological pathogens on the molecular and immunological levels was achieved by a combination of the following enabling technologies:
1. Nano- and microfabrication technologies: Nano- and microfabrication technologies enabled a miniaturization of the system, a massively reduced sample and reagent consumption, a decrease in the overall reaction time, the integration of several biological steps, a parallel processing of two detection reactions without any manual handling steps after loading of the sample.
2. Innovative detection technologies: Electrochemical sensors, based on fabrication strategies developed in the course of the project were directly integrated on the plastic chip and used for the detection on the immunological and molecular levels in an extremely sensitive manner.
3. Biological process steps: A prerequisite for the detection bacterial DNA, which must also be very specific to avoid false positive reactions, is the adequate construction of highly specific DNA primers and probes. Suitable reagents were constructed and validated for the following six bacterial agents of category A&B: Francisella tularensis, Yersinia pestis, Brucella melitensis, Burkholderia mallei, Burkholderia pseudomallei and Coxiella burnetii.
4. Scientific instrumentation: Advanced technologies and novel solutions were combined to both final instruments that cover the complete analytical process: control the fluidic, thermal, electronic and optical functions of the lab-on-a-chip device and, as an add-on, covers sample enrichment from air.
The overall system was realized via the combination of all mentioned technologies, merging engineering science with the biological and diagnostic fields, advanced sensor technologies, and scientific instrumentation. This combination of different disciplines to a multidisciplinary network of enabling technologies posed the highest technological challenge.
The overall systems comprise the following functionalities, which are implemented in the following manner:
Air sampling: For the sample uptake from air, the Coriolis system from Bertin was used as a basic ingredient and adopted to the special needs of the project namely a high recovery rate enabled by long sampling and special process conditions: A dry air sampling process and a permanent operation mode for the system were developed and successfully implemented achieving a significantly higher recovery rate.
Swab sampling: For the sample uptake of suspicious components swabs were the method of choice. A special sampling vessel was developed that is prefilled with buffer in which the swab is inserted after sampling. The innovative features are two characteristics: The sampling vessel can directly mounted on the lab-on-a-chip device with the help of an a proper integrated fluidic interface and due to an intrinsic septum, the vessel can be removed without any contamination risk allowing to send a fraction of a sample to other analytical sides. The swab was transferred in a reaction vessel prefilled with sampling buffer.
Liquid samples: Liquid samples were directly gathered in a sampling vessel and mounted to the instrument. After finishing the analysis of the sample, the vessel can be disposed or be sent to other analytical sides.
Sample preparation and concentration: To isolate DNA and antigens and to reduce the overall sampling volume a special sample preparation module was developed that combines concentration and sample preparation. For sampling a certain volume was transferred to the concentration module and the overall volume of the sample was reduced to a defined volume. During this step not only the DNA and antigens for the molecular and immunological assay were isolated but also the sample was concentrated.
Immunological detection: On chip, a standard immune reaction took place. The specific antibodies were presented in two manners: In the Permanent Monitoring System (PMT) they were immobilized on filter frits that allow for a sample concentration since the complete sample is flushed through these frits as porous elements catching the respective target molecules. In the One-time Measurement System (OTM) the antigens were attached to the gold surface of the electrochemical detection electrodes and the sample was flushed over the array.
Detection on molecular level: After the sample preparation for the DNA pathogens, a PCR (Polymerase Chain Reaction) for the amplification of the target DNA was carried out using a continuous flow approach in the Permanent Monitoring System and a stationary approach in the One-time Measurement System.
Electrochemical detection: This detection technology was used in the One-time Measurement System and required an additional detection reaction in order to generate an electrochemical signal. The electrochemical set-up was read out by electrochemical sensors. For this assay for both detection levels, the molecular and immunological detection, an enzyme label and substrate were employed and the generated product was detected using an novel integrated pluri-potentiostat coping with 64 parallel detections.
Fluorescence detection: Fluorescence detection was used in the Permanent Monitoring System. This detection method required a fluorescence label that was integrated in the DNA during the PCR.
Photometer detection: The last detection method was used to detect the antigens in the Permanent Monitoring System. The basic function is equal to the electrochemical detection. Detection antibody were immobilized on a filtration frit after binding the antigen a detection antibody followed by a detection solution were exposed to the frits. In case of positive detection the color change (reduction of light transmission) of the frit was measured using a photometer.
IT – Software & communication: Efforts on the IT side were made on three levels: First of all, the control software for the instruments to run the chip was successfully established on both instruments. The second level was the software for the read-out of the detection reaction. The third level was the information and communication software that was developed and implemented to the system.
System integration: All functionalities of the instrument ranging from air collection, fluid and thermal management, optical and electronic read-out, data analysis and the communication tool to e.g. first responders and others authorities were successfully integrated into both systems, the Permanent Monitoring System and the One-time Measurement System.
Finally two integrated systems were at hand to cope with the Multisense Chip project goal, namely to analyze biological samples in a sample-to-answer fashion without any manual interaction besides the sample introduction. This demonstrates that besides the technical challenge of the realization of the consumable lab-on-a-chip device and the respective instrument all biological tasks were combined on the platform and that the multidisciplinary team achieved a working system that was successfully evaluated with first responders.

Project Results:
The “Multisense Chip Analyzer” is an easy to handle but technologically complex system combining a multidisciplinary approach from engineering, science and information and communication technology in order to cope with a blead-to-read fashion of the analysis of biological samples. This required development on technical level in order to realize the lab-on-a-chip system and the respective instrument, on biological level to establish all steps from sample uptake over sample preparation to the final assay, and on the communication level to allow for spreading the analytical results in a predefined network of first responders according to a fixed communication chain. Finally, ethical aspects were covered in detail keeping the later user scenario touching the diagnostic area in mind: Critical items such as the potential use of animal or human samples were discussed and respective documents were put in place. The evaluation of the system revealed, however, that neither human nor animal material was used. Furthermore, the handling of restricted information and hazardous substances was covered within the ethical work package.
In order to cope with this multidisciplinary challenge a consequent risk minimization strategy was implemented. Core element of this risk minimization were two elements: A modular approach and standardization: Basis of the modular approach was the division of the overall task in technical module and sub-modules allowing for a parallel development e.g. of single biological steps, evaluating instrument and fluidic behavior separately from the biology etc. In addition, parts of the instrument as well as the different detection technologies were realized and evaluated in parallel. This ensured a concerted development of all biological process steps on technical and biological level. Apart from the modular approach, a standardization concept was implemented defining outer geometries of the lab-on-a-chip device and principle functional elements of the required consumables and interfaces to the instrument. This enabled a parallel development of instrument modules and the lab-on-a-chip device.
Enormous effort on the S&T level in the different areas ended up not only in two integrated systems but also in a wide variety of innovations that are on their way to commercial use. This covers the fields of sensor technology, biological assay development, microfluidic devices and systems as well as respective instruments, the communication technology and finally system integration and system validation. The interdependency of the different tasks was a organizational challenge in order to ensure the parallel efficient development and to draw conclusions from e.g. biological results for the microfluidic device choice.
S&T area ASSAY DEVELOPMENT WP 2: Assay definition and assay development was the starting point, since with the definition of the assays themselves, the targets, the antibodies and first process chain of the biological steps was set. A biological flow chart was developed that was transferred in a fluidic flow chart defining sequences, volumes, timing and thermal or electrical actions.
The biological processes were all developed on chip and for benchmarking the gold standard technology was applied.
In brief, the biological tasks were completely implemented on chip covering sample enrichment, clean-up as well as the analytical detections on immunological and molecular level in a sensitive and quick manner, e.g. by decreasing single analytical steps from hours to minutes by the miniaturized approach. The topic of reagent storage in dry and liquid format was addressed successfully. Single results of this work will be exploited as well as the final implemented assay that will take further development work.
S&T area SENSOR TECHNOLOGY WP 3:
The development of a sensitive sensing technology based on electrochemical or electrochemiluminescence detection to be embedded on chip was the goal. Different work aspects needed to be covered: First task was the electrode development for both sensing technologies covering various design cycles. Secondly, new chemistries were set-up addressing the linker of the catcher molecules to be immobilized on the electrode surface, the material to cover the remaining electrode surface called “back-filler” to avoid unspecific binding, methods to amplify a signal either by multiplying through release of encapsulated particles and working using the enhancing effect of nanoparticles. The implementation of the process of screen-printed as well as lithographically made electrodes and their benchmark was a major task with the achieved aim to work with screen-printed electrodes that can be fabricated in a cost-efficient manner.
In brief: With the help of electrochemical- and electrochemiluminescence-tester set-ups both technologies were implemented successfully, both on screen-printed and lithographically made surfaces. Finally, a great sensitivity was achieved combined with even better results for the screen printed electrodes based on cheap manufacturing technologies and in the biological assay no cross-talk in the arrays with all targets embedded demonstrating the maturity of the technological result chosen.
S&T area MICRO- AND NANOFABRICATION TECHNOLOGIES WP 4:
According to the fabrication chain for the realization of polymer based lab-on-a-chip systems, several fabrication steps needed to be addressed in detail, mainly covering assembly technologies, surface functionalization and electrode fabrication.
Major achievements are the direct integration of the electrodes as hybrid elements working on an adhesive-free assembly principle and an adhesive-based assembling technology of hybrid elements made from different materials in polymer-based devices making use of a fluidic network. Immobilization technologies and respective chemical procedures allowed for a well-defined fabrication of the protein- and DNA-arrays by spotting. The most important outcome was the novel approach for the screen-printed electrodes: Moving from one-sided electrode array to double sided electrode structuring was achieved by a novel technology for implementing contracts through the electrode back-bone material. This technology can be implemented in a cost-efficient manufacturing manner and allows for a massive shrinkage of the electrode footprint, decreasing spotting cost by the order of the size reduction of the electrode device and chip cost through a smaller chip device.
S&T area MICROFLUIDICS AND MICROSYSTEM INTEGRATION WP 5:
Development of microfluidic modules and accessories was key for the later integration of all task on one lab-on-a-chip platform. Therefore, results for a straight forward exploitation are lab-on-a-chip modules for the three implemented PCR technologies, sample preparation units, sampling vessels and fluidic interfaces.
Special highlights are both integrated chips, the Multisense Chip design 14 (MUSE 14) for the Permanent Measurement System and Multisense Chip design 15 (MUSE 15) for the One Time Measurement System. All biological steps were implemented successfully ending up with fluorescence and colorimetric detection for the Permanent Measurement System and an electrochemical detection for the Permanent Measurement System.
S&T area SOFTWARE, COMMUNICATION & INFORMATION MANAGEMENT WP 6:
The development of the communication module was also successfully achieved and implemented. Three different communication strategies namely Ethernet, GSM and satellite based communication were integrated. A major achievement was the final integration of the satellite communication and having access to such communication tool. Satellite modem proved to be easy to handle, even if not as easy as S-band device as envisaged with the first choice provider that could not be further addressed during the course of the project.
The integration and test of the ICT platform was performed successfully. The communication module proved to handle messages correctly through all communication channels planned (Ethernet, GSM, and satellite). Also, hardware and communication channel controls proved to work efficiently as well as error handling so that messages are still correctly sent, if a piece of hardware, software, or network is down. The integration on the embedded control PC and other components was solved including user interface for configuration of the communication parameters.
S&T area INSTRUMENTATION AND SYSTEM INTEGRATION WP 7:
Modules as building blocks covering PCR, sample preparation, sample concentration, air sampling and various detection technologies were developed, validated and the most appropriate ones were integrated in the final systems. The integration of these modular building blocks beginning with the air sampling module over the EC-tester setup and two different PCR approaches was accomplished within the Permanent Monitoring System (PMS) and the One-Time-Measurement-System (OTM).
The PMS for automated sampling and pathogen detection has the following key features: integrated air sampling module, cooled reagent storage, sample concentration module, continuous flow PCR module, fluorescence based DNA detection, light transmission based antigen detection, embedded PC with touch screen control, and communication module for communication via GSM, Ethernet and satellite.
The OTM for on-site pathogen detection from liquid and swab samples has the following key features: sample concentration module, reagent storage on chip, waste management on chip, electrochemical detection of DNA and antigens, and embedded PC controlled via touch screen.
The fluidic protocols developed for the permanent monitoring and on-time detection chips were transferred into automated control protocols for both instruments. These protocols were tested fluidically and with biological samples and reagents. Furthermore, the information transfer from the analysis software to the operator of the system has been developed and evaluated.
S&T area complete process implementation: VALIDATION, END-USER EVALUATION (WP 8) and DEMONSTRATION OF PRE-PRODUCTION PROTOTYP (WP 9):
Basis for the integration of the complete process was the validation of all single technical modules and the implemented biological procedures.
Field and end-user were integral part of the work plan: Field test with the air sampling module and demonstration of the communication technology were carried out successfully also mimicking different scenarios like human environment in sport settings or in dusty surroundings in stables. The microfluidic modules such as the PCR module for continuous flow and stationary PCR, the sample preparation module for magnetic bead-based on-chip sample preparation, the sample preparation module by thermal concentration, or the air sampling module were validated, which allowed for optimized integration of these modules in the entire lab-on-a-chip system.
Both integrated instruments, the Permanent Monitoring System and the One-time Measurement System, with their appropriately integrated microfluidic chips Multisense design 14 (MUSE 14) and Multisense design 15 (MUSE 15). Although some issues on the Multisense design 15 (MUSE 15) were revealed during the electrochemical detection validation, it could be shown that both chips gained comparable results for reliability and reproducibility as the single modules. The overcoming of the remaining identified issues has to be a major task for the product development phase of both systems.
The validation of the physical parameters of the instruments and chips was accomplished: Robust reaction conditions are of prime importance for a reliable and reproducible detection result. It was demonstrated that the instruments in combination with the chips provide such stable conditions.
Validation of the control software and the communication module showed no issues the software and the detection protocols run reliable and were easy to use.
An end-user demonstration event took place evaluating the handiness and functionality of the Multisense Chip Permanent Monitoring System and the Multisense Chip One-Time Measurement System. End users from the “Brigade de Sapeurs-Pompiers de Paris” and the “Wehrwissenschafliches Institut für Schutztechnologien, Munster” had the opportunity to test extensively and define the list of topics to be improved for both systems.
It was stressed that the test results were correct, there were no false positive detections and the overall system worked.
During the tests several issues regarding performance and handiness of both instruments could be addressed transferred in the recommendation list for improvement combined with a positive feedback of the end users. The acceptance of the end users was high. And the end-users feedback has been very fruitful and will be the base for the product development phase.
In summary, the project goal was reached resulting in two integrated system covering the application scenario of permanent and one-time measurement. The final integrated systems combine hardware, software, fluidic performance and assay implementation aspects and were successfully validated to function in a sample-to-answer manner for the analysis of bacterial pathogens on immunological and molecular level.

Potential Impact:
Potential impact: The potential impact covers first of all the direct use of the developments carried out within the project for the target and other applications as well as the technologies that were established. Gathered reputation and economic results are further impact that needs to be stressed. Due to the untypically high and prominent representation of female scientists in the project, a special socio-economic impact can be reported.

Impact of the systems and their application
The two systems developed within the project, namely the “Multisense Chip Analyzers” for one-time and permanent measurements have a significant impact on how biological analysis, be it for the safety and security scenario, environmental and food screening of medical diagnostic can be carried out.
Due to the fact that with these systems a lab-free-approach to analyze biological samples – usually complex, time consuming, requiring lab space and trained staff – is at hand. Taking the permanent monitoring system that samples air over defined periods, this system has an impact in the security and safety area e.g. for critical infrastructure monitoring, in stadiums, metro stations, air ports or moving in the civilian application to monitor climate control units e.g. in hospitals seeking for critical and usually multi resistant bacteria.
The one-time measurement system allows to quickly analyze suspicious samples at the place of interest and can be placed in critical infrastructure, post offices, airports etc. wherever this material might come up. Expanding the use and sample matrices the medical diagnostic market has a great potential, enabling test labs as well as the respective units of hospital or even the practitioner to carry out the analysis. A future scenario is e.g. home testing of the users health status. Obviously, the impact on the health market are great, despite this requires a major development effort.
Other application in food and environmental testing would require special developments as well but in particular contamination testing would be of major interest. Targeted development projects would quickly allow to develop solutions for this area.
Impact on safety & security & international traffic
The envisaged benefit of the project is the provision of versatile B-detection system to enforce the European safety and security actions and the tools being available.
In particular under the current threat scenario of the IS and availability of technologies to culture and distribute biological agents, the member states are urgently needing such detection systems as developed within Multisense Chip.
Taking a different user scenario into account, internationalization and upcoming epidemic illnesses like Dengue, Zika or Yellow fever virus induced ones can make use of the same technological base.
Impact of the system in a wider scope
Taking the grand challenges of an aging society into account, home testing of the health status combining the lab-on-a-chip approach with the communication tools is a future, challenging but in particular important future impact. This obviously will require enormous effort to the ease of use, system size and cost, consumable cost and the use of smart phone in combination with health apps and communication strategies. Enabling tools were developed within the scope of Multisense Chip.

Impact of the technologies
The technologies that were developed will be included in lab-on-a-chip systems with increased performance e.g. through novel reagent implementation options, sensor integration technologies or new fluidic building blocks used to developed the integrated cartridges. Both involved SMEs (iMicroQ and MFCS) and their customers and partners will benefit from these developments.
Impact on reputation & actions deriving thereof
Due to the fact that the project brought the team members in the positions, to present their work the relevant players, on a long term view recognition of the partners was achieved. This led e.g. to the participating of MFCS in the expert group (PASAG – Protection and Security Advisory Group) of the European Commission to give advice for the follow-up Research program of Horizon 2020.
The active dissemination activities gave e.g. MFCS an awareness that the company and a system based on work carried out within Multisense Chip was shown on Germany TV celebrating the German Reunification as example of “Eastern Success Stories” showing the promised “Blühende Landschaften” (Prosperous Landscapes) from old chancellor Kohl.

Impact on the economy
The project enabled the involved industry to develop new products, build know-how and benefit from a multidisciplinary consortium.
Finally, it is envisaged to achieve a double digit Mio. Euro annual turnover based on the outcome of the project. E.g. “Multisense Chip” allowed MFCS to create new business and was one of the key enabler to employ 10 more team members in the company group, partly for the project itself, but mainly to cover new topics deriving from the project’s outcome. From project application to finalization of the project, MFCS has doubled its staff also demonstrating the economic impact of the technology and the effect of EU financial support on international competitiveness. Currently two spin-off-companies are in discussion that will benefit from Multisense Chip outcome.

Impact for Europe
The benefit for Europe through Multisense Chip matures on three levels, starting with scientific excellence, over the economic impact to finally the Multisense Chip products covering the security and safety related perspective. And the achievements already made, nicely demonstrate that the funding provided was fruitfully used to achieve the promised goals.

Impact – wider societal implications
Apart from these results related to the work done in the project, this project had an additional impact as side effect. Both, project coordinator and project manager as well as other leading scientist at the partners URV and FLI were female. This is still completely unusual in a technical field – in particular in a ‘high tech’ area. This project coped with an outstanding way to the still interesting item of gender issues via allowing this team not only to prove to run this project successfully but also enabled the female scientists to present the work at high level events for international audience predominated by the usual gender distribution. With the help of this project it was clearly shown on an international floor, how Europe supports women and their impact in science and business. For girls and young women, who have been addressed by the project as well e.g. via “Girls’ Days”, it gives an insight in an innovative technology field and an impression how fascination for science can be turned into fun, scientific excellence, and economic success leading to more young women in science.
Furthermore, with Multisense Chip the project coordinator Dr. Claudia Gärtner was two times finalist in the Women Innovator Competition of the European Commission and Dr. Gärtner was decorated in 2014 with the Emily-Röbling-Prize for female business power.

Main dissemination activities
The complete bundle of dissemination tools was applied during Multisense Chip: Starting with the creation of an Exploitation Committee followed by the preparation of dissemination templates, the project homepage and material like flyers and posters a roadmap how to identify and address the respective players and create business from the project’s outcome was defined. Major methods to create awareness were conference, congresses and trade fairs combined with scientific publications. MFCS achieved own Multisense Chip sessions at one of the major conferences of the SPIE dealing with safety and security. Working with interest groups and joining panel discussions on the safety and security topic lead to a well appreciated awareness of the projects and the coordinator MFCS as a known player in this field. More than 20 participating in relevant exhibitions and trade fairs, more than 45 oral presentations, 8 peer reviewed papers, 25 published papers, posters and activities addressing the local population demonstrate the active work in the dissemination area.
Special highlights of the dissemination events were being panelist of the event to present the “Masterplan civilian security economy” of the German Ministry of Economy and Technology, the Panel discussion on “Flagship EU-projects” during the Millipol 2013 and the EDA conference “Security Matters” 2014. Special visibility created the presence in the IB-Consulting series on NCT (Non conventional threats) CBRNE that resulted in awareness and partners for distribution and promotion of the Multisense Chip results.

Exploitation of results
Exploitation of results happened on four levels taking also the different kind partners into account: The first level has been the creation of awareness, reputation and collaboration being relevant for all partners: This included publications, being member of interest groups, getting awareness as player in the field, contact to distribution partners or partners for further collaborations and follow-up R&D projects. All partners have been active on this level. Also the second level “Know-how” has been exploited by all partners in respect to new technological and application know-how. “Products”, described as exploitation level three, has been the most visual outcome of the project, the relevant level all industrial partners benefit from. The fourth level has been the “End-user” view represented by FLI who will finally use the results.
The exploitation of results has already started, a general exploitation scope can be given and some prominent examples are highlighted.

General exploitation scope
The final results of the project were two analytical systems covering the detection of biological pathogens as a permanent monitoring system and as a one-time measurement system.
These systems channel the way to an improved, easier, and faster detection of biological pathogens and will enable responders and authorities to quickly get aware of potential contaminations and to be able to decide on potential contaminations with such agents leading to a possibility for a fast and appropriate measure. Far beyond this security driven application, veterinarian and medical diagnostics will benefit from such systems in long term.
Besides the complete system, also modules thereof such as improved air sampling system with added functionalities, and a set of microfluidic toolbox components, e.g. lab-on-a-chip consumables and instruments for sample preparation, enzyme assays, PCR etc., visualize the results of the project.
Both integrated systems and modules have the potential to impact laboratory or analytical routine. A major outcome is that in future untrained personnel can operate complex analytical tasks allowing for a new pathway to operated risk situation not only in the security but in particular to address outbreak scenarios in the veterinarian field.
The integration level provided by the communication and information aspect covered in this project will further facilitate the use of the system in critical environments and will give an automatism at hand, how analytical results will be transferred into actions, such as generating a message and starting a communication process directly by the “Multisense Chip Analyzer”.
Next to the direct products that came out of the project, their use for the partners involved in this project and the future users, the network established within this project has been of major benefit for the partners. The SMEs benefit from the interdisciplinary know-how provided the access to special infrastructure at the institutes, the experiences in hardening and validation of Bertin as larger company and their existing network, whereas Bertin gets access to a new technology and potential novel products.
Projects making use of the outcome of Multisense Chip
The created awareness through the Multisense Chip project turned into the partnering in two R&D projects within the European framework and an EDA-Contract: The FP 7 project EDEN: Achievements of the Multisense Chip fluidic design was used to create further designs and to be able to join a demo in the food area in April 2016. A Horizon 2020 project: Project to start mid 2016 were the PCR fluidic part will be complemented with a real time optical detection. A development contract for the EDA (European Defence Agency) called RAMBO combines the real time PCR to be read-out optically with a RAMAN-based detection. The project is ongoing. For this project a Bertin air sampler is in use.
Current plans for development with external partners are a Dengue and Yellow Fever platform based on work carried out within Multisense Chip.
Module – side products with direct exploitation impact
In order to realize the overall integrated system a modular approach was chosen to minimize the development risk and being able to develop single units in parallel. The outcome of this building block-work are several new products: A blister test-platform for liquid storage, a lab-on-a-chip mini-thermocycler with and without real-time detection and a novel multipotentiostat for a 64 channel. Furthermore, a novel air sampler option and sample preparation units for sample concentration and communication tools are exploitable on the short term from the companies involved.

Impact, dissemination & exploitation – summary
Due the fact that the Multisense Chip project was constantly represented at conferences, exhibitions and public events a high level of awareness for the project could be brought up
In particular the activities to address the CBRNE community were accelerated, e.g. with a participation in the CBRNE meeting in Berlin in October 2012 or presentations on the SPIE Security and Defense meeting in May 2013 in Baltimore. Further events in 2013 as the CBRNE ASIA or the Medical B-Defence conference followed. Besides the directly CBRNE focused events Multisense Chip results have been presented on numerous trade fairs partners being present.
For the exploitation in the CBRNE field, the integrated instruments are the final exploitable results. Starting with test users the return on invest will mature on a longer time frame compared to the exploitation of technologies and sub-modules that can be used for various applications.
Publications are important for exploitation and dissemination as well. Various publications were done. Besides conference presentations, several proceeding papers and peer reviewed papers were published. This included e.g. several papers and posters at the SPIE Defense, Security & Sensing or the SPIE Photonic West.
The fact that exploitation took place on different levels, ensured that all partners got a return on invest and that the risk of complete failure was minimized. In particular the SMEs profited from technologies and modules that could be exploited shortly after the project end resulting in a broader product and technology range, increased turnover, and the creation of new job.
The networking within the consortium and via contacts to further groups and networks mediated by other partners was strengthened. All involved parties were able to create new contacts and improve already existing ones for future collaboration on R&D as well as commercial level.
New R&D projects and contacts to distributors are the direct outcome initiated through the Multisense Chip project that will be a base for the commercialization of the integrated Multisense Chip products.

List of Websites:
www.multisense-chip.com
Contact: Dr. Claudia Gärtner
microfluidic ChipShop GmbH, Stockholmer Str. 20 , 07747 Jena

Final Report Summary - MULTISENSE CHIP (The lab-free CBRN detection device for the identification of biological pathogens on nucleic acid and immunological level as lab-on-a-chip system applying multisensor technologies)

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