Microfluidics is an interdisciplinary research field that deals with the production and the study of droplets on a micrometric scale. In the last decades microfluidics has gained an increasing interest by the scientific community. Microfluidics has arisen great attention since it controls the transport of small amounts of liquids inside which, for example, it is possible to make chemical reactions occur or through which is possible to transport biological samples or diagnostic markers. Their features, in combination with fast analysis tools, allow for the realization of Lab-on-a-Chips (LOCs) that is to say miniaturized and portable devices able to perform chemical, biological, environmental or medical analyses where bulky laboratory facilities lack. As a matter of fact in microfluidics' standards, imaging techniques are exploited to monitor the droplets generation and/or movement. In these cases, however, bulky microscopes eliminate the obtained advantages with miniaturization. For this reason, it is important to integrate completely on a small scale the optical stage with the microfluidic one achieving a so called opto-microfluidic platform, an objective accomplished in the last few years, for example, through the realization of Lab-On-a-Chip in Lithium Niobate in the Physics and Astronomy department "Galileo Galilei", in Padua. This material has excellent optical properties that consent to host the realization of Ti-indiffused waveguides, by which it is possible to detect the produced droplets and their contents in the engraved microfluidic channel as it will be shown in this thesis. This thesis presents a feasibility study on the use of integrated optics coupled to microfluidics circuitry to investigate the proteins synthesis. Since the process of the protein synthesis could be difficult and require several days, it is important to save as much as possible of the biological samples, by periodically monitoring its concentration both after the final purification process, and during the subsequent experiments. However, this usually implies the use (and waste) of a significant amount of the sample, so that often a compromise between these two aspects has to been found. On the contrary, in principle the use of the opto-microfluidic approach presented in this thesis would allow to comply with both the mentioned needs, since the monitoring of the protein concentration would be realized in a fast ways and without requiring large amount of the biological samples. Moreover, it can allow to monitor the effect of each update that can be introduced to the process to improve the synthesis or add extra elements into the proteins. The work-plan of the thesis includes the realization and validation of the opto-microfluidic platform and the application to the detection of proteins. It consists of the characterization of various experimental setups until reaching the best configuration to monitor droplets containing proteins and a reagent called Bradford through a completely integrated opto-microfluidic device in Lithium Niobate. The study is aimed at understanding how proteins' synthesis can be monitored even in small amounts as some confined in a micrometric droplet. The thesis aims to demonstrate that, by investigating how the protein-based droplet interacts with a light beam. It will be demonstrated that depending on the measurement of the optical transmission of such a droplet it is possible to detect if and when the protein has been synthesized. The final aim of the project, this thesis is included in, is the obtainment of a qualitative proteins detector. The presence of the proteins in the dispersed phase is evaluated by means of the transmitted light using a systematic measurement protocol and having good control on the experimental apparatus thanks to the detailed characterization. The results obtained show that it is possible for our setup to detect qualitatively the presence of proteins in micrometric droplets.

Opto-microfluidic device in Lithium Niobate to investigate the proteins synthesis ​

Turato, Enrico
2021/2022

Abstract

Microfluidics is an interdisciplinary research field that deals with the production and the study of droplets on a micrometric scale. In the last decades microfluidics has gained an increasing interest by the scientific community. Microfluidics has arisen great attention since it controls the transport of small amounts of liquids inside which, for example, it is possible to make chemical reactions occur or through which is possible to transport biological samples or diagnostic markers. Their features, in combination with fast analysis tools, allow for the realization of Lab-on-a-Chips (LOCs) that is to say miniaturized and portable devices able to perform chemical, biological, environmental or medical analyses where bulky laboratory facilities lack. As a matter of fact in microfluidics' standards, imaging techniques are exploited to monitor the droplets generation and/or movement. In these cases, however, bulky microscopes eliminate the obtained advantages with miniaturization. For this reason, it is important to integrate completely on a small scale the optical stage with the microfluidic one achieving a so called opto-microfluidic platform, an objective accomplished in the last few years, for example, through the realization of Lab-On-a-Chip in Lithium Niobate in the Physics and Astronomy department "Galileo Galilei", in Padua. This material has excellent optical properties that consent to host the realization of Ti-indiffused waveguides, by which it is possible to detect the produced droplets and their contents in the engraved microfluidic channel as it will be shown in this thesis. This thesis presents a feasibility study on the use of integrated optics coupled to microfluidics circuitry to investigate the proteins synthesis. Since the process of the protein synthesis could be difficult and require several days, it is important to save as much as possible of the biological samples, by periodically monitoring its concentration both after the final purification process, and during the subsequent experiments. However, this usually implies the use (and waste) of a significant amount of the sample, so that often a compromise between these two aspects has to been found. On the contrary, in principle the use of the opto-microfluidic approach presented in this thesis would allow to comply with both the mentioned needs, since the monitoring of the protein concentration would be realized in a fast ways and without requiring large amount of the biological samples. Moreover, it can allow to monitor the effect of each update that can be introduced to the process to improve the synthesis or add extra elements into the proteins. The work-plan of the thesis includes the realization and validation of the opto-microfluidic platform and the application to the detection of proteins. It consists of the characterization of various experimental setups until reaching the best configuration to monitor droplets containing proteins and a reagent called Bradford through a completely integrated opto-microfluidic device in Lithium Niobate. The study is aimed at understanding how proteins' synthesis can be monitored even in small amounts as some confined in a micrometric droplet. The thesis aims to demonstrate that, by investigating how the protein-based droplet interacts with a light beam. It will be demonstrated that depending on the measurement of the optical transmission of such a droplet it is possible to detect if and when the protein has been synthesized. The final aim of the project, this thesis is included in, is the obtainment of a qualitative proteins detector. The presence of the proteins in the dispersed phase is evaluated by means of the transmitted light using a systematic measurement protocol and having good control on the experimental apparatus thanks to the detailed characterization. The results obtained show that it is possible for our setup to detect qualitatively the presence of proteins in micrometric droplets.
2021-04
92
Lab-On-a-Chip, Lithium Niobate, Microfluidics, Protein synthesis, Laser, Bradford, Bovine serum albumine ​
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12608/28714