(1201-B) A Multi-purpose Automation System for Large-scale Super-resolution Image Data Acquisition

Abstract: In recent years, there has been remarkable progress in super-resolution microscopy (SRM). While SRM has made it possible to observe cellular dynamics at the molecular scale, artificial intelligence (AI) technology has been gaining attention as a means of mining novel knowledge which is difficult to detect by manual observation. The combination of SRM and AI is expected to provide unprecedented insights into many applications for cellular biology. However, combining SRM and AI raises several issues. One of the issues is a sample preparation. SRM often requires carefully prepared quality samples which is currently done manually. On the other hand, AI requires a large amount of evenly quality data which means that the manual preparation of samples has high risk of failure or misconduct. To address this issue and realize the SRM-AI combination, the authors developed an automated super-resolution imaging system; Automated Sample preparation and image Acquisition Platform (ASAP). ASAP combines a general-purpose bio-experiment automation system called LabDroid Maholo (RBI) and a spinning disk laser confocal super-resolution microscope called CSU-W1 SoRa (Yokogawa). This system enables us to acquire a large amount of quality image data required for AI analyses by automating both sample preparation and image acquisition. Samples for SRM are prepared by Maholo using a glass-bottom microplate, then the microplate is transported, by a robot arm, to an integrated plate stacker which can store more than 10 microplates waiting for image acquisition. Automated sample transport feature is integrated into NIS-Elements AR equipped with a custom dynamic link library (DLL) which communicates with the robot controller (Intel PC w/ CODESYS v3) to transport a microplate between the stacker and the microscope stage. During microplate transport, barcode scanning is performed for automated sample management. This sample transport command can be issued from JOBS using a macro function which enables microscope users to easily utilize the automated sample transport for their image acquisitions. CSU-W1 SoRa is equipped with two Fusion-BT (Hamamatsu) sCMOS cameras enabling two-color synchronous imaging. We are currently working on a task to acquire a large amount of super-resolution two-color Z-stack Golgi body images for clarifying cell-cycle dependent Golgi body morphological dynamics (https://projectdb.jst.go.jp/grant/JST-PROJECT-21466156/). In addition to the fluorescent SRM, our microscope is capable of high magnification (x100) apodised phase contrast microscopy which enables stain-free live imaging at super-resolution with reduced phase artifacts. ASAP is designed for multi-purpose image acquisition for various kinds of samples which makes ASAP unique among similar automation systems.