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NanoJ-Fluidics


GitHub:
- HenriquesLab/NanoJ-Fluidics

Publication: Almada et al. Nature Communications 2019
- 86

NanoJ-Fluidics is an automated, computer-controlled system for exchanging fluids at the microscope sample during imaging experiments. It allows researchers to perform complex multimodal microscopy protocols with high reproducibility and minimal manual intervention.

The system consists of an array of compact Lego syringe pumps that can be multiplexed to automatically dispense reagents onto the sample. The hardware is controlled by open-source ImageJ-based software, enabling precise timing of fluid delivery. Key capabilities of NanoJ-Fluidics include:

  1. Event-driven fixation and staining: The system can fix cells at a specific timepoint based on visual cues during live imaging, allowing researchers to capture dynamic processes. For example, it fixed cells undergoing mitotic rounding by dispensing fixative when cell circularity exceeded a threshold. It then automatically performed immunostaining.

  2. Live-to-fixed correlative microscopy: NanoJ-Fluidics enables in situ correlation of live and fixed cell imaging. It first captures dynamic events like cytoskeletal rearrangements using live super-resolution microscopy. When ready, it fixes the cells, stains them, and switches to fixed-sample super-resolution techniques like STORM. This links nano-scale architecture with functional imaging.

  3. Multiplexed super-resolution imaging: The system exchanges imaging buffers to sequentially perform multi-color STORM and DNA-PAINT. This simplifies difficult protocols and expands the color palette. For example, it achieved 5-color images by interleaving STORM of actin with two rounds of DNA-PAINT staining.

The key advantages of NanoJ-Fluidics are its simplicity, flexibility and integration with imaging. Researchers avoid manual wash steps at the bench. The Lego pumps enable customizable setups for different microscopes. Crucially, the ImageJ-based software allows protocols to be precisely timed and triggered based on analysis of microscopy images. For example, mitotic rounding automatically triggered fixation and immunostaining.

NanoJ-Fluidics makes complex microscopy protocols highly reproducible and accessible. By automating fluid exchange, it opens new possibilities for live-to-fixed correlative imaging and multiplexed super-resolution microscopy. The capacity to trigger events based on microscopy images also enables new biologically-driven experiments. Its simple, open-source nature means that many researchers should be able to incorporate NanoJ-Fluidics into their workflows to enhance multimodal imaging.


Publications featuring NanoJ-Fluidics

The Rise of Data-Driven Microscopy powered by Machine Learning
Leonor Morgado, Estibaliz Gómez-de-Mariscal, Hannah S Heil, Ricardo Henriques
Preprint published in arXiv, January 2024
Technologies: BioImage Model Zoo, DL4MicEverywhere, NanoJ, NanoJ-Fluidics and ZeroCostDL4Mic
Funded by: CZI, EMBO, ERC, FCT, H2021 and H2022
DOI: 10.48550/arXiv.2401.05282
Live-cell imaging in the deep learning era
Joanna W Pylvänäinen, Estibaliz Gómez-de-Mariscal, Ricardo Henriques, Guillaume Jacquemet
Review published in Current Opinion in Cell Biology, January 2023
Technologies: BioImage Model Zoo, CARE, DeepBacs, Fast4DReg, NanoJ, NanoJ-eSRRF, NanoJ-Fluidics, NanoJ-SRRF and ZeroCostDL4Mic
Funded by: CZI, EMBO, ERC and H2021
DOI: 10.1016/j.ceb.2023.102271
The LEGO® brick road to open science and biotechnology
Etienne Boulter, Julien Colombelli, Ricardo Henriques, Chloé C. Féral
Review published in Trends in Biotechnology, March 2022
Technologies: NanoJ, NanoJ-Fluidics and NanoJ-SRRF
Funded by: EMBO, ERC and Wellcome Trust
DOI: 10.1016/j.tibtech.2022.02.003
The field guide to 3D printing in microscopy
Mario Del Rosario, Hannah S Heil, Afonso Mendes, Vittorio Saggiomo, Ricardo Henriques
Review published in Adv. Biol., January 2021
Technologies: CARE, NanoJ, NanoJ-Fluidics and NanoJ-SRRF
Funded by: EMBO, ERC and Wellcome Trust
DOI: 10.1002/adbi.202100994
The cell biologist's guide to super-resolution microscopy
Guillaume Jacquemet, Alexandre F. Carisey, Hellyeh Hamidi, Ricardo Henriques, Christophe Leterrier
Review published in Journal of Cell Science, June 2020
Technologies: CARE, NanoJ, NanoJ-Fluidics, NanoJ-SRRF, Nuclear-Pores as references and ZeroCostDL4Mic
Funded by: BBSRC and Wellcome Trust
News: ScienMag and EurekAlert!
DOI: 10.1242/jcs.240713
Between life and death - strategies to reduce phototoxicity in super-resolution microscopy
Kalina L Tosheva, Yue Yuan, Pedro Matos Pereira, Siân Culley, Ricardo Henriques
Review published in Journal of Physics D - Applied Physics, January 2020
Technologies: CARE, NanoJ, NanoJ-Fluidics and NanoJ-SRRF
Funded by: BBSRC and Wellcome Trust
DOI: 10.1088/1361-6463/ab6b95
An Introduction to Live-Cell Super-Resolution Imaging
Siân Culley, Pedro Matos Pereira, Romain F Laine, Ricardo Henriques
Book chapter published in Imaging from Cells to Animals In Vivo, January 2020
Technologies: CARE, NanoJ, NanoJ-Fluidics, NanoJ-SQUIRREL, NanoJ-SRRF and QuickPALM
DOI: 10.1201/9781315174662-4
Super‐beacons - Open‐source probes with spontaneous tuneable blinking compatible with live‐cell super‐resolution microscopy
Pedro M Pereira, Nils Gustafsson, Mark Marsh, Musa M Mhlanga, Ricardo Henriques
Paper published in Traffic, January 2020
Technologies: NanoJ, NanoJ-Fluidics, NanoJ-SQUIRREL, NanoJ-SRRF and Super-Beacons
Funded by: BBSRC and Wellcome Trust
DOI: 10.1111/tra.12728
Artificial intelligence for microscopy - what you should know
Lucas von Chamier, Romain F. Laine, Ricardo Henriques
Review published in Biochemical Society Transactions, July 2019
Technologies: CARE, NanoJ, NanoJ-Fluidics, NanoJ-SQUIRREL and NanoJ-SRRF
Funded by: BBSRC and Wellcome Trust
DOI: 10.1042/bst20180391
Automating multimodal microscopy with NanoJ-Fluidics
Pedro Almada, Pedro M. Pereira, Siân Culley, Ghislaine Caillol, Fanny Boroni-Rueda, Christina L. Dix, Guillaume Charras, Buzz Baum, Romain F. Laine, Christophe Leterrier, Ricardo Henriques
Paper published in Nature Communications, March 2019
Technologies: NanoJ, NanoJ-Fluidics, NanoJ-SQUIRREL, NanoJ-SRRF and NanoJ-VirusMapper
Funded by: BBSRC and Wellcome Trust
News: Technology Times, MSN, DNYUZ and Express Informer
DOI: 10.1038/s41467-019-09231-9
NanoJ - a high-performance open-source super-resolution microscopy toolbox
Romain F Laine, Kalina L Tosheva, Nils Gustafsson, Robert D M Gray, Pedro Almada, David Albrecht, Gabriel T Risa, Fredrik Hurtig, Ann-Christin Lindås, Buzz Baum, Jason Mercer, Christophe Leterrier, Pedro M Pereira, Siân Culley, Ricardo Henriques
Paper published in Journal of Physics D - Applied Physics, January 2019
Technologies: NanoJ, NanoJ-Fluidics, NanoJ-SQUIRREL, NanoJ-SRRF, NanoJ-VirusMapper and QuickPALM
Funded by: BBSRC and Wellcome Trust
DOI: 10.1088/1361-6463/ab0261
Fix your membrane receptor imaging - actin cytoskeleton and CD4 membrane organization disruption by chemical fixation
Pedro M Pereira, David Albrecht, Siân Culley, Caron Jacobs, Mark Marsh, Jason Mercer, Ricardo Henriques
Published in Frontiers in immunology, January 2019
Technologies: CARE, NanoJ, NanoJ-Fluidics and NanoJ-SQUIRREL
Funded by: BBSRC and Wellcome Trust
DOI: 10.3389/fimmu.2019.00675

Funding contributing to NanoJ-Fluidics

Sub-cellular Metabolic Compartmentalization During Oocyte Development
Zita Carvalho dos Santos, Ricardo Henriques, Jorge Carvalho
Funded by: CZI - Measuring Metabolism Across Scales
Duration: January 2024 - December 2026
A transformative data-driven live-cell super-resolution microscopy development to elucidate the initial steps of effective viral transmission
Estibaliz Gómez-de-Mariscal
Funded by: EMBO - Postdoctoral Fellowships
Duration: July 2022 - June 2024
Publications: 5
Real-Time high-content Super-Resolution Imaging of ES Cell States
Eran Meshorer, Ricardo Henriques, Anna Kreshuk, Sandrine Lévêque-Fort, Nicolas Bourg, Genevieve Almouzni
Alias: RT-SuperES
Funded by: H2022 - EIC Pathfinder Open
Duration: April 2022 - March 2027
Publications: 6
VP-CLEM-KIT - a pipeline for democratising volumetric visual proteomics
Lucy Collinson, Ricardo Henriques, Paul French
Funded by: CZI - Visual Proteomics Imaging
Duration: December 2021 - June 2024
Publications: 11
Enabling Live-Cell 4D Super-Resolution Microscopy Guided by Artificial Intelligence
Ricardo Henriques
Alias: SelfDriving4DSR
Funded by: ERC - Consolidator
Duration: September 2021 - September 2026
Publications: 23
Mapping the early stages of HIV-1 infection by live-cell 4D Super-Resolution Microscopy
Hannah Heil
Funded by: EMBO - Postdoctoral Fellowships
Duration: September 2021 - August 2023
Publications: 5
Optial Biology PhD programme
Michael Hausser, Ricardo Henriques, Antonella Riccio
Funded by: Wellcome Trust - 4-year PhD Programme in Science
Duration: August 2021 - August 2025
Mapping HIV-1 infection by 4D Super-Resolution Microscopy
Hannah Heil
Funded by: FCT - CEEC Individual
Duration: July 2021 - June 2027
Publications: 2
Unveiling live-cell viral replication at the nanoscale
Ricardo Henriques
Funded by: EMBO - Installation Grant
Duration: January 2021 - January 2026
Publications: 16
4D Super-Resolution Proteomics
Ricardo Henriques
Funded by: UCL - Capital Equipment Fund
Duration: June 2019 - May 2020