An Emerging Remote Sensing An Emerging Remote Sensing Technology and its Potential Technology and its Potential

Impact on Mine ActionImpact on Mine Action

GeoTec GmbH International Symposium Humanitarian Demining 2010, Sibenik, Croatia International Symposium Humanitarian Demining 2010, Sibenik, Croatia

What we propose:What we propose: To establish a new remote sensing technology suitable for the To establish a new remote sensing technology suitable for the

stand off detection of trace explosives leaking as a tell tale signal stand off detection of trace explosives leaking as a tell tale signal from landmines of all types and builds. from landmines of all types and builds.

For that we join for the first time two high technologies …..For that we join for the first time two high technologies ….. We will employ microbiology to produce a genetically engineered We will employ microbiology to produce a genetically engineered

but safe soil bacterium to use it as a sensitive fluorescent live but safe soil bacterium to use it as a sensitive fluorescent live biosensorbiosensor for trace explosives. for trace explosives.

We will use commercially available We will use commercially available airborne laser technology airborne laser technology (LIDAR)(LIDAR) and modify it to and modify it to searchsearch rapidly and over wide areas for rapidly and over wide areas for fluorescent biosensors on the ground indicating the presence and fluorescent biosensors on the ground indicating the presence and location of landmines.location of landmines.

What are biosensors?What are biosensors? A device that uses biological materials to monitor the presence A device that uses biological materials to monitor the presence

of various chemicals in a substance. of various chemicals in a substance. This could be for example an enzyme-based or antibody-based This could be for example an enzyme-based or antibody-based

sensor “mounted” in a “lab on a chip” using some opto-electric sensor “mounted” in a “lab on a chip” using some opto-electric device for the read out.device for the read out.

What are What are livelive biosensors? biosensors? A living organism or a A living organism or a single living cellsingle living cell capable to sense and capable to sense and

signal by itself the presence of a substance in the environment signal by itself the presence of a substance in the environment it is moving in.it is moving in.

How can a single living cell be How can a single living cell be made to signal something we can made to signal something we can

then detect?then detect? By using the naturally acquired and genetically encoded capability of a By using the naturally acquired and genetically encoded capability of a

cell to sense and either utilize or avoid a substance, and …cell to sense and either utilize or avoid a substance, and … By genetically tying the sensing process to a second and artificially By genetically tying the sensing process to a second and artificially

introduced biological signaling process which makes the cell produce introduced biological signaling process which makes the cell produce fluorescent proteins as a signal detectable for an observer.fluorescent proteins as a signal detectable for an observer.

Bacteria under a Bacteria under a microscope labeled microscope labeled with green and red with green and red fluorescent proteinsfluorescent proteins

Braincells of a mouse Braincells of a mouse labeled with labeled with fluorescent proteinsfluorescent proteins

Fluorescent miceFluorescent mice

What is an airborne LIDAR?What is an airborne LIDAR? The acronym LIDAR stands for The acronym LIDAR stands for LiLight ght DDetection etection aand nd RRanginganging Airborne LIDAR systems integrate a pulsed laser scanner, a Global Airborne LIDAR systems integrate a pulsed laser scanner, a Global

Positioning System (GPS) and an Inertial Measurement Unit (IMU).Positioning System (GPS) and an Inertial Measurement Unit (IMU). A modern airborne LIDAR sends typically about 100.000 ultra A modern airborne LIDAR sends typically about 100.000 ultra

short laser pulses per second to the ground. It measures the total short laser pulses per second to the ground. It measures the total traveltime of each individual light pulse from the LIDAR to the traveltime of each individual light pulse from the LIDAR to the ground and back as a reflection to the receiver unit of the LIDAR. ground and back as a reflection to the receiver unit of the LIDAR. Depending on the flight altitude and flight speed 5 – 10 precision Depending on the flight altitude and flight speed 5 – 10 precision distance measurements per square meter are sampled. distance measurements per square meter are sampled.

Using ultra precise navigation giving the position and orientation Using ultra precise navigation giving the position and orientation of the aircraft for each time a laser pulse was fired to the ground of the aircraft for each time a laser pulse was fired to the ground the system is capable to calculate the position and distance of the system is capable to calculate the position and distance of each laser reflection point on the ground. These data are then each laser reflection point on the ground. These data are then used to generate Digital Elevation Models describing the overflown used to generate Digital Elevation Models describing the overflown land surface with great detail. Digital Elevation Models can be land surface with great detail. Digital Elevation Models can be worked with using normal office computers or lap topsworked with using normal office computers or lap tops

LIDARS were initially developed for military applications like target LIDARS were initially developed for military applications like target acquisition and precision navigation of cruise missiles. A special acquisition and precision navigation of cruise missiles. A special application was submarine hunting using green laser light. We application was submarine hunting using green laser light. We eventually will use such a system and modify it to hunt landmines.eventually will use such a system and modify it to hunt landmines.

Civilian applications nowadays are for example the precision Civilian applications nowadays are for example the precision mapping of urban areas, flood plane mapping, corridor planning mapping of urban areas, flood plane mapping, corridor planning and open pit mining.and open pit mining.

Digital Elevation Model SamplesDigital Elevation Model Samples

Elevations are color codedElevations are color coded Three versions of the same model stackedThree versions of the same model stacked

Digital Elevation Model SamplesDigital Elevation Model Samples

A LIDAR can penetrate the canopy of a forest and in fact detect and A LIDAR can penetrate the canopy of a forest and in fact detect and look at the ground!!!look at the ground!!!

Golden ruleGolden rule: if an observer in a forest can see small patches of the : if an observer in a forest can see small patches of the sky above him the LIDAR can see the ground – and of course sky above him the LIDAR can see the ground – and of course fluorescent biosensors on the ground!fluorescent biosensors on the ground!

Specialized soil bacteria like Pseudomonas putida can use explosives Specialized soil bacteria like Pseudomonas putida can use explosives as their carbon source.They consider TNT as a tasty food. These as their carbon source.They consider TNT as a tasty food. These bacteria can be genetically engineered to produce fluorescent bacteria can be genetically engineered to produce fluorescent proteins in the cell envelop if they detect and consume traces of TNT proteins in the cell envelop if they detect and consume traces of TNT or DNT.or DNT.

Engineered and TNT Engineered and TNT sensitive Pseudomonas sensitive Pseudomonas putida soil bacteria putida soil bacteria encapsulated in encapsulated in alginate pellets. They alginate pellets. They did already produce red did already produce red fluorescent proteins fluorescent proteins after exposure to traces after exposure to traces of TNTof TNT.

Excitation of fluorescent Excitation of fluorescent proteins in biosensors using proteins in biosensors using a green laser to induce a red a green laser to induce a red fluorescent response. Only fluorescent response. Only biosensors which had biosensors which had contact with TNTand contact with TNTand produced fluorescent produced fluorescent proteins will responde with proteins will responde with red fluorescence.red fluorescence.

Resulting red fluorescence Resulting red fluorescence in the biosensors shown on in the biosensors shown on the left. The response time the left. The response time to exitation with green to exitation with green laser light is laser light is instantaniously (speed of instantaniously (speed of light) which allows us to light) which allows us to detect this process in full detect this process in full flight.flight.

back to the back to the Biosensor..Biosensor..

How to use this technology to detect How to use this technology to detect hazardous areas requiring full hazardous areas requiring full

clearance and how to find single clearance and how to find single landmines?landmines? Mix freeze dried starter cultures of biosensors with water to obtain Mix freeze dried starter cultures of biosensors with water to obtain

a sprayable solution of functional biosensors. This can be done in a sprayable solution of functional biosensors. This can be done in the field at the airstrip using clean water, clean barrels and some the field at the airstrip using clean water, clean barrels and some nutritional salts. This process is likely to take several hours.nutritional salts. This process is likely to take several hours.

Spray biosensors over wide areas taking care that no space is left Spray biosensors over wide areas taking care that no space is left untreated. Then wait a few hours for biosensors to interact with untreated. Then wait a few hours for biosensors to interact with their environment and produce fluorescent proteins in case they their environment and produce fluorescent proteins in case they sensed trace explosives. sensed trace explosives.

Fly sensor plane with mounted LIDAR to excite, detect and Fly sensor plane with mounted LIDAR to excite, detect and georeference any fluorescent biosensors on the ground. georeference any fluorescent biosensors on the ground.

After landing of the plane produce Digital Elevation Model (DEM) After landing of the plane produce Digital Elevation Model (DEM) to show exact positions of fluorescent response from the ground. to show exact positions of fluorescent response from the ground. Hopefully the fluorescence on the ground copies the extent of a Hopefully the fluorescence on the ground copies the extent of a Halo of trace explosives in the soil next to a single landmine.Halo of trace explosives in the soil next to a single landmine.

Digital Elevation Model generated by the LIDAR doubles as high Digital Elevation Model generated by the LIDAR doubles as high precision GIS to assist in all further activities.precision GIS to assist in all further activities.

How can biosensors be spread How can biosensors be spread quickly over wide areas of quickly over wide areas of

interest?interest?

Use a crop duster with precision flight management system.Use a crop duster with precision flight management system. The daily capacity of a single crop duster plane is yet unknown for our The daily capacity of a single crop duster plane is yet unknown for our

application. It would depend on the required density of biosensors on the application. It would depend on the required density of biosensors on the ground and the packaging of biosensor (weight).ground and the packaging of biosensor (weight).

Shown plane (AirTractor) has a payload of 3,5 mt and is capable to perform 80 Shown plane (AirTractor) has a payload of 3,5 mt and is capable to perform 80 starts and landings per day. Estimated daily capacity 30 square kilometers.starts and landings per day. Estimated daily capacity 30 square kilometers.

How can billions of fluorescent How can billions of fluorescent biosensors on the ground be detected biosensors on the ground be detected

from the air and in full flight?from the air and in full flight?

A pulsed laser system (LIDAR) scans and measures the surface using a A pulsed laser system (LIDAR) scans and measures the surface using a green laser to generate a Digital Elevation Model. green laser to generate a Digital Elevation Model.

The same green laser light is simultaniously used to excite, detect and The same green laser light is simultaniously used to excite, detect and georeference red fluorescent light from biosensors activated by trace georeference red fluorescent light from biosensors activated by trace explosives. explosives.

Localisation precision of fluorescence: better than 0,5 m in the horizontal Localisation precision of fluorescence: better than 0,5 m in the horizontal plane.plane.

Flight altitude 800 meters, swath width 200m, daily capacity 100 square Flight altitude 800 meters, swath width 200m, daily capacity 100 square kilometers!!kilometers!!

Measuring fluorescence intensity Measuring fluorescence intensity with a LIDARwith a LIDAR

Shown is a Digital Elevation Model of an airstrip.Shown is a Digital Elevation Model of an airstrip. In this case no color coded elevations are shown but gray coded In this case no color coded elevations are shown but gray coded

intensities of the reflected laser light entering the receiver.intensities of the reflected laser light entering the receiver. The painted surfaces on the strip yield the highest reflection The painted surfaces on the strip yield the highest reflection

intensities.intensities. This functionality of a LIDAR can be used to detect fluorescent This functionality of a LIDAR can be used to detect fluorescent

light from biosensors and its intensities.light from biosensors and its intensities.

A color coded Digital Elevation A color coded Digital Elevation Model for the visual orientation of Model for the visual orientation of

the deminer on the groundthe deminer on the ground

The same digital elevation model with The same digital elevation model with „photoshopped“ signals from „photoshopped“ signals from

fluorescent biosensors on the ground fluorescent biosensors on the ground indicating the positions of landminesindicating the positions of landmines

Preliminary results after two years of Preliminary results after two years of researchresearch

BiosensorBiosensor:: A functional and safe fluorescent biosensor for traces of TNT and A functional and safe fluorescent biosensor for traces of TNT and

DNT wasDNT was developed and tested in bio-security labs at developed and tested in bio-security labs at Fraunhofer Institute for Microbiology in Germany.Fraunhofer Institute for Microbiology in Germany.

The biosensor is based on the begnin soil bacterium The biosensor is based on the begnin soil bacterium Pseudomonas putida Pseudomonas putida and produces a large amount of bright red and produces a large amount of bright red fluorescent proteins upon contact with TNT or DNT.fluorescent proteins upon contact with TNT or DNT.

The biosensor is sensitive enough to detect TNT and DNT in The biosensor is sensitive enough to detect TNT and DNT in concentrations between 1ppm – 10 ppm dissolved in liquids and concentrations between 1ppm – 10 ppm dissolved in liquids and 10 ppm dissolved in soil (1 10 ppm dissolved in soil (1 ppm==1mg/kg==1mg/ltr.) ppm==1mg/kg==1mg/ltr.)

This is in the range of reported TNT and DNT This is in the range of reported TNT and DNT concentrations found in soils next to landminesconcentrations found in soils next to landmines. .

As our biosensors settle on the soil surface we might benefit from As our biosensors settle on the soil surface we might benefit from a process where TNT molecules dissolved in water get enriched a process where TNT molecules dissolved in water get enriched at the surface in dried up puddles.at the surface in dried up puddles.

Development of a laboratory scale laser scanner set up in a dedicated Development of a laboratory scale laser scanner set up in a dedicated biosafety laboratory at the Fraunhofer Institute for Lasertechnology in biosafety laboratory at the Fraunhofer Institute for Lasertechnology in Germany. Germany.

The laser scanner was used to test and calibrate the genetically produced The laser scanner was used to test and calibrate the genetically produced live fluorescent biosensor for TNT and DNT.live fluorescent biosensor for TNT and DNT.

Dectetion of the live fluorescent biosensor in the lab over a distance of 12 Dectetion of the live fluorescent biosensor in the lab over a distance of 12 meters (we consider that already a safe stand off distance).meters (we consider that already a safe stand off distance).

Development of a mock up biosensor exhibiting the same spectral response Development of a mock up biosensor exhibiting the same spectral response as the genetically modified live biosensor. This mock up biosensor could be as the genetically modified live biosensor. This mock up biosensor could be used without any restrictions outside of a biosafety lab.used without any restrictions outside of a biosafety lab.

Outdoor tests of the mock up biosensor and detection of small volumes of Outdoor tests of the mock up biosensor and detection of small volumes of mock up biosensors over a distance of 300 meters using a stationary mock up biosensors over a distance of 300 meters using a stationary mounted airborne fluorescence LIDAR employed by the German mounted airborne fluorescence LIDAR employed by the German government in patrol flights over German costal waters to detect oil spills. government in patrol flights over German costal waters to detect oil spills.

Proof of principle for the airborne detection of fluorescent Proof of principle for the airborne detection of fluorescent biosensors.biosensors.

Preliminary results after two years of Preliminary results after two years of researchresearch

Laser scannerLaser scanner::