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Adding A New Experimental Parameter

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0. Design questions before you start

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Does the new experimental parameter require sensing, actuation, or both?

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Examples

  • Sensing only: optical sensor (photodiode, phototransistor) for new spectra, pH sensor

  • Actuation only: optical stimulation w/ new LED

  • Both: fluorescence measurements (LED/optical sensor pair), feedback driven optical stimulation

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Additional Questions

  • Can new hardware be fitted onto vial?

    • Hardware CAD files can be found .

  • Can the eVOLVER provide necessary power to control actuator?

The best way to answer these questions is to go through the datasheets of components you are interested in integrating and breadboarding with the eVOLVER Motherboard. This will help you also establish and tune dynamic ranges for new sensing capabilities. Additionally, you can explore swapping in new power supplies for components with higher power consumption specs. Once you've established compatibility and feasibility, you can begin integrating a new experimental parameter for eVOLVER.

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Low Level (Hardware/Arduino)

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1. Add appropriate boards (PWM/ADC) to SA slots in eVOLVER Motherboard.

  • Each Arduino is connected to 2 SA slots so that culture parameters that require sensing/actuation can share data, but Arduino is still capable of controlling each SA slot individually.

  • If parameter requires only sensing or actuation, then its best to pair with parameters that also require sensing or actuation (i.e. stir) for efficient usage of Arduinos

  • If the experimental parameter is sensing a culture condition, plug in a ADC Board

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2. Program Arduino to collect serial data and/or regulate actuators.

  • All Arduinos communicate with Raspberry Pi along same serial line and either look for tags/add tags associated with the parameter they’re responsible for controlling

  • GitHub and wiki more information regarding how to program Arduinos

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3. Add new hardware components to Smart Sleeve

  • Smart Sleeve 3D printed housing may need to be redesigned to fit components

  • Solder component leads to correct Component Mount Board (CMB) pins, which is set by slot position of SA slot(s)

    • E.g. If using SA slot 8, make sure that components leads are wired to position 8 on CMB

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High Level (Server/DPU)

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4. Add experimental parameter to eVOLVER server configuration file (conf.yml) with relevant configurations

  • If parameter is for actuating, 'fields_outgoing' set to 17 (see example below):

  • If parameter is for sensing, fields_outgoing set to 2 (see example below):

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5. Adjust DPU software to take advantage of new experimental parameter in continuous culture algorithms

  • of wiki goes in more depth on how to create culture control algorthms

  • Basically, you need to create a new method in the EvolverNamespace class, which can be called in your custom control code

  • Example below using default stir parameter:

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Make sure that the name you are using for the experimental parameter is consistent throughout this process. eVOLVER will pull a list of experimental parameters from the conf.yml to sort through incoming serial data and tag outgoing serial commands. Inconsistencies with naming will lead to issues later on!

The base eVOLVER can provide 5V or 12V signal to the vials.

  • Otherwise, look into changing the .

  • Is the signal you are sensing strong enough to be sensed using the components you will use?

  • If the experimental parameter is applying a stimulus to the culture, plug in a PWM Board

    herearrow-up-right
    repoarrow-up-right
    docs
    DPU section
      temp:
        recurring: true
        fields_expected_outgoing: 17
        fields_expected_incoming: 17
        value: ['4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095', '4095']
    experimental_params:
      od_90:
        recurring: true
        fields_expected_outgoing: 2
        fields_expected_incoming: 17
        value: '1000'
    def update_stir_rate(self, stir_rates, immediate = False):    
        data = {'param': 'stir', 'value': stir_rates, 
        'immediate': immediate, 'recurring': True}    
        logger.debug('stir rate command: %s' % data)    
        self.emit('command', data, namespace = '/dpu-evolver')
    PWMarrow-up-right
    power supply

    Power Supply

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    Relevant Forum Posts

    About the current power supplyarrow-up-right
    Clarification on Power Supplyarrow-up-right

    Specific Applications

    Adding pH and Dissolved Oxygen Sensorsarrow-up-right

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    Optogenetic Control During Continuous Culture

    Forum Postarrow-up-right

    Light inducible protein domains have been used to dynamically control and rapidly prototype genetic networks24,52. Hardware for light inducible systems typically rely on batch culture, limiting experiments to a narrow time window in which all cells across an experiment are in exponential phase. Attempts at coupling continuous culture to light induction have been limited by throughput (1-2 cultures) and reconfigurability. Equipped with components for light induction, eVOLVER would uniquely enable long-term optogenetic perturbations in finely controlled growth phases across a large number of culture vessels. Due to the modularity of eVOLVER hardware components, integrating optogenetic control is straight forward and requires minor modifications to the system. Details on integrating LEDs into eVOLVER is described in more detail as the example modification in Supplementary Note 4.

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    Fluorescence Measurements

    Bulk fluorescence measurements have previously been demonstrated by Takahashi et al. during continuous culture, without the use of a photomultiplier tube (PMT)7. To recapitulate this setup in eVOLVER, an extra LED-diode pair would be added to the 6th and 7th S/A Slots, similar to adding LEDs for light induction. Additionally, the 3D printed part would be modified to house optical filters for better detection of any fluorescence signal. Potential setbacks in this setup (without a PMT) include potentially a low signal to noise ratio.

    This can be solved by multiplexing signal from all cultures into a single PMT via fiber optics. The electronics controlling the PMT would communicate back to the same RS485 line to be controlled by the same Raspberry Pi, similar to the auxiliary board. Alternatively, single cell fluorescence measurements would also be made possible by interfacing eVOLVER with a pipetting robot, droplet microfluidics, or using the native pump from the flow cytometer sample directly from the cultures. These systems could interface serially with the Raspberry Pi via RS485/USB or the lab computer via USB.

    Custom Calibration Code