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Openspritzer: an open hardware pressure ejection system for reliably delivering picolitre volumes

C.J. Forman, H. Tomes, B. Mbobo, View ORCID ProfileT. Baden, View ORCID ProfileJ.V. Raimondo
doi: https://doi.org/10.1101/093633
C.J. Forman
1School of Life Sciences, University of Sussex, United Kingdom
4TReND in Africa gUG ()
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  • For correspondence: chrisforman@cantab.net t.baden@sussex.ac.uk joseph.raimondo@uct.ac.za
H. Tomes
2Division of Physiology, Department of Human Biology, Neuroscience Institute and Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa
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B. Mbobo
2Division of Physiology, Department of Human Biology, Neuroscience Institute and Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa
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T. Baden
1School of Life Sciences, University of Sussex, United Kingdom
3Institute for Ophthalmic Research, University of Tuebingen, Germany
4TReND in Africa gUG ()
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  • For correspondence: chrisforman@cantab.net t.baden@sussex.ac.uk joseph.raimondo@uct.ac.za
J.V. Raimondo
2Division of Physiology, Department of Human Biology, Neuroscience Institute and Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa
4TReND in Africa gUG ()
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  • For correspondence: chrisforman@cantab.net t.baden@sussex.ac.uk joseph.raimondo@uct.ac.za
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Abstract

The ability to reliably and precisely deliver picolitre volumes is an important component of biological research. Here we describe a high-performance, low-cost, open hardware pressure ejection system (Openspritzer), which can be constructed from off the shelf components. When connected to a standard micro-pipette via suitable pneumatic tubing, the device is capable of delivering minute doses of reagents to a wide range of biological and chemical systems. In this work, we characterise the performance of the device and compare it to a popular commercial system using two-photon fluorescence microscopy. We found that Openspritzer provides the same level of control over delivered reagent dose as the commercial system. Next, we demonstrate the utility of Openspritzer in a series of standard neurobiological applications. First, we used Openspritzer to deliver precise amounts of the neurotransmitters glutamate and GABA to hippocampal neurons to elicit time- and dose-precise excitatory and inhibitory responses, respectively. Second, we used Openspritzer to deliver infectious viral and bacterial agents to living tissue. Viral transfection of hippocampal interneurons with channelrhodopsin allowed for the optogenetic manipulation of hippocampal circuitry with light. Finally, we successfully used Openspritzer to infect organotypic hippocampal slice cultures with fluorescent Mycobacterium bovis bacilli. We anticipate that due to its high performance and low cost Openspritzer will be of interest to a broad range of researchers working in the life and physical sciences.

  • List of abbreviations

    ACSF
    Artificial Cerebro Spinal Fluid
    BOM
    Bill of Materials
    CRISPR
    Clustered Regularly Interspaced Short Palindromic Repeats
    DPDT
    Double Pole Double Throw
    EBSS
    Earle’s Balanced Salt Solution
    EPSP
    Excitatory Post Synaptic Potential
    GABA
    γ-aminobutyric acid
    GAD2
    Glutamic Acid Decarboxylase type 2
    GFP
    Green Fluorescent Protein
    GTP
    Guanosine triphosphate
    HEPES
    4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
    LED
    Light Emitting Diode
    MEM
    Minimal Essential Medium
    SPST
    Single Pole Single Throw
    TTL
    Transistor-Transistor logic
    YFP
    Yellow Fluorescent Protein
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    Posted December 13, 2016.
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    Openspritzer: an open hardware pressure ejection system for reliably delivering picolitre volumes
    C.J. Forman, H. Tomes, B. Mbobo, T. Baden, J.V. Raimondo
    bioRxiv 093633; doi: https://doi.org/10.1101/093633
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    Openspritzer: an open hardware pressure ejection system for reliably delivering picolitre volumes
    C.J. Forman, H. Tomes, B. Mbobo, T. Baden, J.V. Raimondo
    bioRxiv 093633; doi: https://doi.org/10.1101/093633

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