Automating multimodal microscopy with NanoJ-Fluidics

• Published in: bioRxiv
• Main Authors: Almada, Pedro, Pereira, Pedro, Si n Culley, Caillol, Ghislaine, Boroni-Rueda, Fanny, Dix, Christina L, Laine, Romain F, Charras, Guillaume, Baum, Buzz, Leterrier, Christophe, Henriques, Ricardo
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 14.05.2018
• Subjects: Actin; Automation; Labeling; Microscopy; Mitosis
• DOI: 10.1101/320416

Fluorescence microscopy can reveal all aspects of cellular mechanisms, from molecular details to dynamics, thanks to approaches such as super-resolution and live-cell imaging. Each of its modalities requires specific sample preparation and imaging conditions to obtain high-quality, artefact-free images, ultimately providing complementary information. Combining and multiplexing microscopy approaches is crucial to understand cellular events, but requires elaborate workflows involving multiple sample preparation steps. We present a robust fluidics approach to automate complex sequences of treatment, labelling and imaging of live and fixed cells. Our open-source NanoJ-Fluidics system is based on low-cost LEGO hardware controlled by ImageJ-based software and can be directly adapted to any microscope, providing easy-to-implement high-content, multimodal imaging with high reproducibility. We demonstrate its capacity to carry out complex sequences of experiments such as super-resolved live-to-fixed imaging to study actin dynamics; highly-multiplexed STORM and DNA-PAINT acquisitions of multiple targets; and event-driven fixation microscopy to study the role of adhesion contacts in mitosis.