WP6 Patho-Stick and Patho-doc development and prototyping General Assembly Meeting WP6 Patho-Stick and Patho-doc development and prototyping

Objectives Design, blueprint and prototyping : Validating functions Patho-stick (UCBL) Patho-doc (Bee) Validating functions Preparation of the pre-industrial specifications

Period achievements Two technical meetings Patho-stick: phone conference BEE, UCBL  Technical_Meeting_Stick_phone_17-07-2015 Extraction: in Molzyme premises (Bremen) MOL, UCBL  Technical_Meeting_Extraction_Bremen_21_22-07-2015 List of requirements for stick and doc Blueprint of patho-doc Prototyping patho-stick

List of requierements (D6.1) Parameter Options Impact Status DNA extraction systems Blood volume used 1mL Size of patho-doc, volume of reagents, sensitivity 1 mL Plastic for consumable Printed or molded Price/material/number of vials Printed but transferable to molding. Polyacrylate for printing. Use of the patho-stick for which sample Blood, urine, Broncho washing, feaces From Swap. Sampling, Extraction, Reagent Blood for development steps, urine in a second time NO SWAP Assay design Quantitative approach YES/NO All WP YES Detection principle Steps and detection tools Particles, colorimetry, fluorescent Colorimetry (enzyme or particles) Amplification Classical PCR or isothermal Patho-doc functions Isothermal To be confirm by AIT Surface chemistry Glass, polystyrene Material availability, probe modification Polystyrene

List of requierements (D6.1) Patho-stick Compatibility between stick configuration for extraction and assay YE/NO Complexity of production   YES, the same stick configuration shall be used for extraction and assay. Only changing volumes and reagents Size 10-100mm Printing technology 15cm*5cm*5cm (500mL bottle size) 3D organization Flat Layered Assembly Either Stick pre-filled Liquid reagent Solid reagents Valves and protocol Yes, as possible Both maybe Array measurement In the stick Outside the stick Transparency of at least one material Integrated measurement Detector in the stick or in the doc Technological complexity of the stick Optical detector on the patho-doc Size of the array Permit naked eye reading 200µm with 600µm pitch at least Price of the patho-stick Material cost around 2€-20€.

List of requierements (D6.1) Patho-doc Footprint   Patho-doc footprint is likely to be approximately 300x350mm Pipetting Which volume Type of liquid handling system involved assay and extraction shall work using 0.5-1mL range volume Function to be implemented Pipetting, heating, cooling, lightning, filtration, imaging… Complexity of the machine Heating 18°C-90°C (hopefully isothermal) Imaging Moving parts (rotating device is foreseen) Filtrate under vacuum (600 mBar) Sterilization Possibility of machine sterilization Materials used YES, UV is foreseen Waste container In the stick or bellow Size of the stick Bellow (12mL) To be confirmed.

List of requierements (D6.1) Prototyping duty Patho-stick for extraction and assay production and prototyping duty   UCBL and MOL for determination of extraction protocol UCBL and Bee for prototype models UCBL for prototyping

Blueprint patho-doc Patho-doc & Patho-Stick Conceptual designs Compact instrument Suitable for running both Extraction and Micro-Array protocols Simple interface Connection to external PC for interpretation of results

Blueprint patho-sticks Patho-doc & Patho-Stick Conceptual designs Patho-Stick (Extraction) Moulded carousel component for 1ml extraction protocol. Positions for reagents and buffers to process single sample. Heated position for mixing (W0). Vacuum cartridge is separate to patho-stick. Patho-Stick (Micro-Array Assay) Separate moulded carousel component Heated reaction plate position. Reaction plate to be a separate transparent moulded part, suitable for imaging.

Blueprint patho-doc Patho-doc & Patho-Stick Conceptual designs 5 Axis instrument Proven linear actuators and drives Stepper motor control

Blueprint patho-doc Patho-doc & Patho-Stick Conceptual designs Conceptual design work is ongoing.

Patho-stick prototyping One stick for assay One stick for extraction One through for Patho-Robot