Vicente Vieira Rocha

Abstract
Optical tweezers are an interesting tool to enable single cell analysis, especially when coupled with optical sensing and advanced computational methods. Nevertheless, such approaches are still hindered by system operation variability, and reduced amount of data, resulting in performance degradation when addressing new data sets. In this manuscript, we describe the deployment of an automatic and intelligent optical tweezers setup, capable of trapping, manipulating, and analyzing the physical properties of individual microscopic particles in an automatic and autonomous manner, at a rate of 4 particle per min, without user intervention. Reproducibility of particle identification with the help of machine learning algorithms is tested both for manual and automatic operation. The forward scattered signal of the trapped PMMA and PS particles was acquired over two days and used to train and test models based on the random forest classifier. With manual operation the system could initially distinguish between PMMA and PS with 90% accuracy. However, when using test datasets acquired on a different day it suffered a loss of accuracy around 24%. On the other hand, the automatic system could classify four types of particles with 79% accuracy maintaining performance (around 1% variation) even when tested with different datasets. Overall, the automated system shows an increased reproducibility and stability of the acquired signals allowing for the confirmation of the proportionality relationship expected between the particle size and its friction coefficient. These results demonstrate that this approach may support the development of future systems with increased throughput and reliability, for biosciences applications.