Single cell manipulation techniques have important applications in laboratory and clinical procedures such as intracytoplasmic sperm injection (ICSI) and polar body biopsy for preimplantation genetic diagnosis (PGD). Conventionally, manipulation of cells conducted in these procedures have been performed manually, which entails long training hours and stringent skills. Conventional single cell manipulation also has the limitation of low success rates and poor reproducibility due to human fatigue and skill variations across operators. This research focuses on the integration of computer vision microscopy and control algorithms into a system for the automation of the following single cell manipulation techniques: (1) sperm immobilization, (2) cell aspiration into a micropipette, and cell positioning inside a micropipette, and (3) rotational control of cells in three dimensions. These automated techniques eliminate the need for significant human involvement and long training. Through experimental trials on live cells, the automated techniques demonstrated high success rates.

Single cell manipulation techniques have important applications in laboratory and clinical procedures such as intracytoplasmic sperm injection (ICSI) and polar body biopsy for preimplantation genetic diagnosis (PGD). Conventionally, manipulation of cells conducted in these procedures have been performed manually, which entails long training hours and stringent skills. Conventional single cell manipulation also has the limitation of low success rates and poor reproducibility due to human fatigue and skill variations across operators. This research focuses on the integration of computer vision microscopy and control algorithms into a system for the automation of the following single cell manipulation techniques: (1) sperm immobilization, (2) cell aspiration into a micropipette, and cell positioning inside a micropipette, and (3) rotational control of cells in three dimensions. These automated techniques eliminate the need for significant human involvement and long training. Through experimental trials on live cells, the automated techniques demonstrated high success rates.

State-of-the-art machine learning methods such as convolutional neural networks (CNNs) are frequently employed in computer vision. Despite their high performance on unseen data, CNNs are often criticized for lacking transparency — that is, providing very limited if any information about the internal decision-making process. In some applications, especially in healthcare, such transparency of algorithms is crucial for end users, as trust in diagnosis and prognosis is important not only for the satisfaction and potential adherence of patients, but also for their health. Explainable artificial intelligence (XAI) aims to open up this “black box,” often perceived as a cryptic and inconceivable algorithm, to increase understanding of the machines’ reasoning.XAI is an emerging field, and techniques for making machine learning explainable are becoming increasingly available. XAI for computer vision mainly focuses on image classification, whereas interpretability in other tasks remains challenging. Here, I examine explainability in computer vision beyond image classification, namely in semantic segmentation and 3D multitarget image regression. This thesis consists of five chapters. In Chapter 1 (Introduction), the background of artificial intelligence (AI), XAI, computer vision, and optics is presented, and the definitions of the terminology for XAI are proposed. Chapter 2 is focused on explaining the predictions of U-Net, a CNN commonly used for semantic image segmentation, and variations of this architecture. To this end, I propose the gradient-weighted class activation mapping for segmentation (Seg-Grad-CAM) method based on the well-known Grad-CAM method for explainable image classification. In Chapter 3, I present the application of deep learning to estimation of optical aberrations in microscopy biodata by identifying the present Zernike aberration modes and their amplitudes. A CNN-based approach PhaseNet can accurately estimate monochromatic aberrations in images of point light sources. I extend this method to objects of complex shapes. In Chapter 4, an approach for explainable 3D multitarget image regression is reported. First, I visualize how the model differentiates the aberration modes using the local interpretable model-agnostic explanations (LIME) method adapted for 3D image classification. Then I “explain,” using LIME modified for multitarget 3D image regression (Image-Reg-LIME), the outputs of the regression model for estimation of the amplitudes. In Chapter 5, the results are discussed in a broader context. The contribution of this thesis is the development of explainability methods for semantic segmentation and 3D multitarget image regression of optical aberrations. The research opens the door for further enhancement of AI’s transparency.:Title Page i List of Figures xi List of Tables xv 1 Introduction 1 1.1 Essential Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.1.1 Artificial intelligence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.1.2 Explainable . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1.1.3 Proposed definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1.2 Explainable Artificial Intelligence . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1.2.1 Aims and applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1.2.2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.3 Computer Vision . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 1.3.1 Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 1.3.2 Image classification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 1.3.3 Image regression . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 1.3.4 Image segmentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 1.4 Optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 1.4.1 Aberrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 1.4.2 Zernike polynomials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 1.5 Thesis Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1.5.1 Motivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1.5.2 Dissertation outline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 2 Explainable Image Segmentation 23 2.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 2.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 2.3 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 2.3.1 CAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 2.3.2 Grad-CAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 2.3.3 U-Net . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 2.3.4 Seg-Grad-CAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 2.4 Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 2.4.1 Circles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 2.4.2 TextureMNIST . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 2.4.3 Cityscapes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 2.5 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 2.5.1 Circles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 2.5.2 TextureMNIST . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 2.5.3 Cityscapes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 2.6 Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 2.7 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 3 Estimation of Aberrations 55 3.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 3.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 3.3 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 3.3.1 PhaseNet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 3.3.2 PhaseNet data generator . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 3.3.3 Retrieval of noise parameters . . . . . . . . . . . . . . . . . . . . . . . . 62 3.3.4 Data generator with phantoms . . . . . . . . . . . . . . . . . . . . . . . 62 3.3.5 Restoration via deconvolution . . . . . . . . . . . . . . . . . . . . . . . . 63 3.3.6 Convolution with the “zero” synthetic PSF . . . . . . . . . . . . . . . . 63 3.4 Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64 3.4.1 Astrocytes (synthetic data) . . . . . . . . . . . . . . . . . . . . . . . . . 65 3.4.2 Fluorescent beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 3.4.3 Drosophila embryo (live sample) . . . . . . . . . . . . . . . . . . . . . . 67 3.4.4 Neurons (fixed sample) . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 3.5 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 3.5.1 Astrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 3.5.2 Conclusions on the results for astrocytes . . . . . . . . . . . . . . . . . . 74 3.5.3 Fluorescent beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 3.5.4 Conclusions on the results for fluorescent beads . . . . . . . . . . . . . . 81 3.5.5 Drosophila embryo . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83 3.5.6 Conclusions on the results for Drosophila embryo . . . . . . . . . . . . . 87 3.5.7 Neurons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88 3.6 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96 4 Explainable Multitarget Image Regression 99 4.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 4.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 4.3 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 4.3.1 LIME . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 4.3.2 Superpixel algorithms . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104 4.3.3 LIME for 3D image classification . . . . . . . . . . . . . . . . . . . . . . 104 4.3.4 Image-Reg-LIME: LIME for 3D image regression . . . . . . . . . . . . . 107 4.4 Results: Classification of Aberrations . . . . . . . . . . . . . . . . . . . . . . . . 109 viii TABLE OF CONTENTS 4.4.1 Transforming the regression task into classification . . . . . . . . . . . . 110 4.4.2 Data augmentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111 4.4.3 Parameter search . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112 4.4.4 Clustering of 3D images . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 4.4.5 Explanations of classification . . . . . . . . . . . . . . . . . . . . . . . . 114 4.4.6 Conclusions on the results for classification . . . . . . . . . . . . . . . . 117 4.5 Results: Explainable Regression of Aberrations . . . . . . . . . . . . . . . . . . 118 4.5.1 Explanations with a reference value . . . . . . . . . . . . . . . . . . . . 121 4.5.2 Validation of explanations . . . . . . . . . . . . . . . . . . . . . . . . . . 122 4.6 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125 5 Conclusions and Outlook 127 References 129

A major difficulty in the fabrication of nanostructure based electronics is the lack of effective processes capable of precisely arranging nanostructures into predefined positions. Top-down approaches introduce increased complexity and a high cost for practical industrial use, while bottom-up approaches are probabilistic in nature and do not provide precise control of nanostructure properties (i.e., number, diameter), which influence device performance. Alternatively, nanomanipulation promises specificity, precision and programmed motion and its automation may facilitate the large-scale fabrication of nanostructure based devices. This study focuses on the development of an automated contact detection algorithm which positions an end-effector in contact with a target surface without the need for additional equipment, devices or sensors. We demonstrate this algorithm as an enabling feature for automated nano-FET biosensor construction with precise control over nanowire parameters thereby reducing device-to-device variability and also potentially allowing us to optimize individual device performance.

A major difficulty in the fabrication of nanostructure based electronics is the lack of effective processes capable of precisely arranging nanostructures into predefined positions. Top-down approaches introduce increased complexity and a high cost for practical industrial use, while bottom-up approaches are probabilistic in nature and do not provide precise control of nanostructure properties (i.e., number, diameter), which influence device performance. Alternatively, nanomanipulation promises specificity, precision and programmed motion and its automation may facilitate the large-scale fabrication of nanostructure based devices. This study focuses on the development of an automated contact detection algorithm which positions an end-effector in contact with a target surface without the need for additional equipment, devices or sensors. We demonstrate this algorithm as an enabling feature for automated nano-FET biosensor construction with precise control over nanowire parameters thereby reducing device-to-device variability and also potentially allowing us to optimize individual device performance.