1 1 Robotic System for In-Vitro Fertilization “Microna IVF-01” Microna Ltd

2 2 Profile of Microna Ltd Microna Ltd is established on 2010 as a spin-off company from the Instiutte of Mechanics. The company patented the technical achievements in the domain of mechatronics and micro-fluidics, integrating them successfully in the field of cell biology and in particular for complex operations with biological cells. Activities Investigation, development and manufacturing of : oRobotic systems and technologies oControl systems and software Marketing and commercialization of products for robotisation of micro & nanotechnologies Consultancy Trainings

3 3 Projects of Microna Ltd (main activity at this stage of development) Name: “Robotic system for in-vitro fertilization” Operative program “Development of the competitiveness of the Bulgarian economy ( )” – Ministry of Economy and Energy Procedure: BG161PO “Development of inovations for starting enterprises” (October 2012 – April 2015). Total amount of the eligible costs: BGN Total abount of the grant: BGN Co-funding: BGN

4 4 Aim of the project Development of a methodology and robotic system for in-vitro fertilization according to ICSI assisted procedure (intra-cytoplasm sperm injection). The improved assisted tool don’t eliminate the embriologist from the whole process but change the skills of the humat with the skills of the robot. Challenges of Microna Ltd Development of an innovative service for in-vitro fertilization, increasing parameters like position accuracy, sterility level, procedure duration. Development of a software for automate control, improving the commercial tele- manipulated systems. Automation of all fluidic processes by a micro-fluidic device. Co-operation with in-vitro clinics in Bulgaria and on abroad, integrating the robotic system in the assisted reproductive medicine.

Сº Z X O Y Z X Y Z X Y X Y Z Design concept of the robotic system for in-vitro fertilization

6 6 Innovative aspect of Microna IVF-01 The system is fully automated and PC controlled The replacement of a Petri-dish with a micro-fluidic device will optimize the control algorithms regarding the velocity and accuracy. Functionality Automate and fast finding of the oocytes in the Petri-dish and learning their coordinates. Positioning of next oocyte in the middles of the working focal space. Automate immobilization of the oocyte by a piezo-manipulator or by a micro-fluidic device. Determination of the orientation of the oocyte, waiting a confirmation for authenticity by operator Assessment of the sperm cells according to shape and velocity, marking of the most suitable and interactive selection by operator.

7 7 Micro-fluidic device: 1-base platform; 2-immobilization module; 3-sperm chamber; 4-oocyte shamber; 5-container for injected cells; 6-bi-directional valve; 7-micro-pump 2; 8-buffer reservoir 2; 9-chamber for the selection module; 10-module for selection and transport; 11-step-moto; 12-buffer reservoir 1; 13-micro-pump 1; 14-micro-hole for immobilisation; 15 – reservoir for non-injected cells Design concept of the micro-fluidic device for ICSI

8 8 Optical system: Automate change of : oObjectives: 5х, 10х, 20х, 40х oFocus: 10nm resolution oIllumination: Digital camera: 5 Mpix, 30 fps, 10 bit Robotic system: 2 piezo-manipulators: Degrees of freedom: - 3 – XYZ Resolution along XYZ: - 1µm Velocity: - up 5mm/sec Range XYZ – 20/20/20mm: XY positioning module: Resolution: - 1µm Range XY: 130/100 mm Micro-fluidic system: Section for sperm cells Section for oocytes: Control: Micro-pumps Functionality o Positioning, immobilization of oocytes; o Release of the injected oocytes. Temperature controlled system: Accuracy: 0.1 C° Computing systems: Work stantion HP Z600 Technical characteristics of Microna IVF-01

9 9 What offers Microna Ltd Selling of the robotic system for in-vitro fertilization Design of such systems, integrating the available module of the client and know- how of the company Services for automated cell manipulations, based on the possibilities of the robotic system for in-vitro fertilization, thus making the manipulations more effective and cheaper (the long work and elimination of the human errors because of the fatigue and loose of concentration). Automation or robotization of processes for testing and validation, different manipulations or injection of cells or other biological objects.

10 Robotisation of the procedure injection of frog oocytes

11 Development, investigation and calibration of a force sensor for measurement of pressure forces over biological cells

12 Development and investigation of MEMS for integration in load cells

13 Experimental measurement of pressure forces over sebrafish embios

14 Thank you for your attention Microna Ltd 1113, Sofia, Bulgaria Acad. G. Bonchev str., bl. 4 Тел.: ,