Joana Magalhães Teixeira

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.

Abstract
Optical trapping provides a way to isolate, manipulate, and probe a wide range of microscopic particles. Moreover, as particle dynamics are strongly affected by their shape and composition, optical tweezers can also be used to identify and classify particles, paving the way for multiple applications such as intelligent microfluidic devices for personalized medicine purposes, or integrated sensing for bioengineering. In this work, we explore the possibility of using properties of the forward scattered radiation of the optical trapping beam to analyze properties of the trapped specimen and deploy an autonomous classification algorithm. For this purpose, we process the signal in the Fourier domain and apply a dimensionality reduction technique using UMAP algorithms, before using the reduced number of features to feed standard machine learning algorithms such as K-nearest neighbors or random forests. Using a stratified 5-fold cross-validation procedure, our results show that the implemented classification strategy allows the identification of particle material with accuracies up to 80%, demonstrating the potential of using signal processing techniques to probe properties of optical trapped particles based on the forward scattered light. Furthermore, preliminary results of an autonomous implementation in a standard experimental optical tweezers setup show similar differentiation capabilities for real-time applications, thus opening some opportunities towards technological applications such as intelligent microfluidic devices and solutions for biochemical and biophysical sensing. © Published under licence by IOP Publishing Ltd.

Abstract
Optical trapping is a versatile and non-invasive technique for single particle manipulation. As such, it can be widely applied in the domains of particle identification and classification and thus used as a tool for monitoring physical and chemical processes. This creates an opportunity for integrating the method seamlessly into optofluidic chips, provided it can be automatized. Yet even though OT is well established in multiple scientific domains, a full stack approach to its integration into other technological devices is still lacking. This calls for solutions in tasks such as automatic trapping and signal analysis. In this manuscript, we describe the implementation of an algorithm seeking autonomous particle location and trapping. The methodology is based upon image-processing, allowing for particle location using real time image segmentation. A local thresholding algorithm is applied, followed by morphological techniques for closing shapes and excluding non-bounded regions - after which only the particles remain on the image. Once the centroid is identified, the stage is translated accordingly by piezo-electric actuators, followed by the laser activation. In this way, trapping is achieved, and one may proceed to analyze the forward scattered optical signal, after which a new particle inside the actuators range may be automatically trapped. This development, when compared with existent solutions involving holographic optical tweezers, allows for similar capabilities without using a spatial light modulator, thus dramatically reducing the setup costs of autonomous OT solutions. Therefore, when combined with particle classification techniques, this method is well suited for integration into possible optofluidic chips for autonomous sensing and monitoring of biochemical samples. © Published under licence by IOP Publishing Ltd.