Lorena Postiglione, M. Biomedical Eng. Tutor: Dr. Diego di Bernardo XXIX Cycle – 1 st year presentation Towards Microfluidics-based Automatic Control of.

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Lorena Postiglione, M. Biomedical Eng. Tutor: Dr. Diego di Bernardo XXIX Cycle – 1 st year presentation Towards Microfluidics-based Automatic Control of gene expression in mammalian cells

Lorena Postiglione 2 Control Engineering in Biology Aim: To force a population of living cells to produce a desired (fixed or time varying) amount of a protein DESIRED fluorescence EXPERIMENTAL fluorescence tetracycline

Microfluidic device and input administration Martin Kolnik et al., Lab on a Chip Inducer Input Untreated Input Syringe Treated medium Syringe Untreated medium Lorena Postiglione 3

4 Experimental Platform tetracycline treated or untreated media

Lorena Postiglione 5 Target reference: 50% of initial steady state fluorescence Calibration time: 3 hrs Control duration: 3500 min Set point control: In vitro control Relay controllerPI-PWM controller

Lorena Postiglione 6 Conclusions & Future Plans Develop model-based controllers Perform tracking control experiment Develop a single cell control strategy DidacticsSeminarsResearch activity 1 st year CFU A feedback control strategy can successfully regulate the gene expression in mammalian cells. Relay and PI controllers don’t achieve optimal performances Automatic control of gene expression in a mammalian cell line Chiara Fracassi, Lorena Postiglione, Gianfranco Fiore, and Diego di Bernardo, ACS Synthetic Biology (in preparation) The Second International Mammalian Synthetic Biology Workshop, April 2015,Massachusetts Institute of Technology Cambridge, MA, USA

Lorena Postiglione 7 Acknowledgements Systems and Synthetic Biology Lab

Lorena Postiglione 8

9 An algorithm computes a mask on the phase contrast image T s =15 min Image acquisition and real-time processing

Controller design: Relay controller with hysteresis u er + system yp - In silico relay control Lorena Postiglione 10

Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM) input uer PI + PWM system yp - Lorena Postiglione 11

Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM) input uer PI + system yp - PWM In silico PI-PWM control Lorena Postiglione 12

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