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Revision as of 09:12, 7 September 2016 by Monika (talk | contribs) (Whole-brain imaging based on OpenSPIM, Warsaw, Poland)

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Aachen, Germany

OpenSPIM with temperature controlled sample chamber by Fraunhofer IPT
  • Life Sciences Engineering Business Unit at the Fraunhofer Institute for Production Technology IPT.
  • Our OpenSPIM design has a temperature controlled sample chamber for long-term experiments and uses a timing belt for precise sample rotation.

Dresden, Germany

  • Tomancak group at the Max-Planck Institute of molecular Cell Biology and Genetics.

Madison, WI, USA

LOCI's L-SPIM, January 22, 2016
  • The OpenSPIM software was developed in this lab by Johannes Schindelin and Luke Stuyvenberg, and tested and refined with the help of Julie Last and Jayne Squirrel (see the People page for details), using that setup. The source code is hosted on GitHub.

Heidelberg, Germany

COS Heidelberg OpenSPIM

Cambridge, UK

Volvox carteri colony imaged with the Goldstein lab OpenSPIM

Melbourne, Australia

OpenSPIM system using the Picard high-resolution stage and an Andor Neo camera in the Petrou Lab, Melbourne, Australia.

Berlin, Germany

OpenSPIM at Humboldt University Berlin
  • The Scholtz group at the Humboldt University Berlin.
  • I am interested in morphogenetic processes during early development of arthropods like the water flea Daphnia magna, the woodlouse Porcellio scaber or the beachhopper Parhyale hawaiensis. Using the SPIM technique for live imaging experiments enables us to observe dynamic and complex processes like gastrulation and beginning morphogenesis.
  • We implemented a Coherent Obis 561 nm laser and a Hamamatsu OrcaFlash4 sCMOS camera into the OpenSPIM system. The system runs at the moment with just one-side illumination. However, the sample chamber is already made for dual-side illumination (2x 10x illumination and 16x detection). Additionally we integrated a Nikon-module that allows us to increases our range of magnification (1x, 1,25x, 1,5x, 2x).
  • More pictures to come!

T-SPIM chamber, designed by Pete Pitrone and build at the workshop of the MPI CBG Dresden.

Exeter, UK

OpenSPIM at the University of Exeter.

Oulu, Finland

OpenSPIM at the University of Oulu.
  • The Light Microscopy Core Facility, at the Biocenter Oulu and the University of Oulu.
  • Modified T-SPIM: dual side illumination with Vortran Stradus 488nm and 561nm lasers, ScopeLed G180 for bright field/transmitted light imaging, Olympus UMPLFLN 20X/0.5 W and Olympus LUMPLFLN 40X/0.8 W detection objectives, Hamamatsu ORCA-Flash 4.0 V2 camera.
  • Temperature, CO2 and O2 controlled liquid exchange/perfusion system for live cell imaging.
  • OpenSPIM has been used for live cell imaging: different cell types and spheroids in different 3D culture matrices, e.g. in Matrigel, collagen, myogel and fibrin mounted inside FEP tubes, and also for imaging living mouse skin cancer biopsies. It has also been used for whole mount stained and/or transgenic (expressing tdTomato, GFP, mCherry…) fixed samples, typically mouse embryonic kidneys, pieces of adult mouse tissues like heart, liver, fat, lung etc.

Indianapolis, IN, USA

IU OpenSPIM chamber with oblique illumination.
  • The Indiana Center for Biological Microscopy at Indiana University School of Medicine.
  • We extended the base L-SPIM design for use in a core facility by including additional excitation wavelengths and support for live imaging. We are imaging with an ORCA Flash 4.0 v2 using CameraLink.
  • To support more fluorophores we incorporated five laser lines (442, 488, 514, 568 and 647 nm) with AOTF modulation controlled via an Arduino based 6 channel DAC. Emission wavelength filtering is with a workhorse Sutter Lambda 10-2. Multichannel acquisition is fully controlled (AOTF, filter wheels, etc) by a custom version of the SPIM Acquisition Plugin and derivations of the Arduino device adapter for MicroManager. To assess laser power, a motorized mirror and ThorLabs detector have been incorporated prior to beam shaping.
  • To support live imaging we have incorporated a perfusion and temperature control system into the standard L-SPIM chamber(no drilling etc.) built with off-the-shelf components (shout-out to Adafruit and Sparkfun). It is Arduino based and relies on a bunch of pumps and thermistors. MicroManager integration and a CO2 loop is in the works (anyone with experience implementing amphenol CO2 sensors?).
  • Our OpenSPIM is fully contained to simplify laser safety. To facilitate alignment and sample positioning a secondary camera driven by a RaspberryPi provides a live image of the chamber with oblique RGB LED illumination from an Arduino and PWM via the 16 channel TLC5940.
  • To aid in sample positioning a knob bar is under development with four free-turning knobs-one for each axis with position feed-back.
  • IU OpenSPIM has been used for imaging of Zebrafish (live heart-beat and fixed and cleared tissue), organoids, and tissue samples including rat kidneys-most in FEP tubing. It has also been a testbed for new technologies and methodologies.

  • Links to schematics, Arduino firmware, PCB designs, MicroManager device adapters coming soon!

Whole-brain imaging based on OpenSPIM, Warsaw, Poland

Cleared-tissue imaging SPIM at the Nencki Institute.
  • We are the Laboratory of Neurobiology at the Nencki Institute of Experimental Biology, Warsaw, Poland [1]
  • Using OpenSPIM as the starting point, we developed a cleared-tissue imaging SPIM system using dry 4X/0.13 Nikon objectives with WD of 17 mm, or about 25 mm in immersion
  • We use 3D printed chambers for CLARITY- and CUBIC-cleared tissue and glass cuvettes for BABB-cleared tissue
  • Recently we replaced the Picard stage with one by Standa, with greatly increased speed and travel range at comparable cost
  • Also recently a second illumination arm was added, converting the setup from the L to the T configuration
  • Ar-Ion laser (458/488/515 nm with manual filter wheel for wavelength selection) used for illumination
  • we have demonstrated whole mouse and rat brain imaging (see paper)

Newcastle (HMRI/UoN), Australia

OpenSPIM HMRI/UoN glamour shot
  • The 3D Tissue Clearing and Lightsheet Microscopy Facility (HMRI/UoN).
  • Our focus is on the development and progression of 3D histology in medical and plant sciences.
  • Imaging is performed with an OpenSPIM-based T-SPIM with Hammamatsu Orca Flash, Arduino controlled 405, 473 and 532nm CNI lasers, and Thorlabs emission filters and dichroics
  • Picard 4D stage controlled by wireless 3D 'spacemouse' - highly recommended.
  • Controlled via Micromanager, with 3D dataprocessing performed with FIJI, Terastitcher and Vaa3D using the UoN HPC Cluster
  • Currently building a new SPIM incorporating a digitally scanned lightsheet (to take advantage of the lightsheet mode of the Orca Flash), two Nikon x4 Plan Fluorite illumination objectives (17mm WD), 1" optics, enlarged chamber, and a long working distance RI corrected detection objective for imaging large, cleared tissues