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eVOLVER
  • eVOLVER Documentation Wiki
  • General
    • About Us
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      • Code of Conduct
  • Getting Started
    • Buying eVOLVER
    • Part Sourcing
    • Unboxing and Setting Up
    • Software Installation
      • DPU Installation
      • Electron App (GUI) Installation
    • Configuring Computer and Networking
      • Router Setup
    • Calibrations
      • Temperature Calibration
      • Optical Density Calibration
      • Pump Calibration
      • Manual Calibration - calibrate.py
  • Upgrade Base eVOLVER Hardware
  • Experiments
    • Starting an Experiment
      • Carboy Media Prep
      • Preparing Vials
      • Setup Waste Carboy
      • Sterilizing Lines
      • Loading Vials and Setting Initial Conditions
      • OD Blank
      • GUI Start Guide
      • Command Line Start Guide
      • Cleaning Up After Experiment
    • Growth Curve
    • Chemostat
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    • FAQs
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  • Guides
    • Use the GUI to Control Parameters
    • Building a Smart Sleeve
    • Making media bottles and splitters
    • Emergency Efflux
    • View the Server Log and Restart Server
    • Updating the eVOLVER Server
    • Change Your conf.yml File
    • Arduino Software Installation
    • Raspberry Pi Configuration
    • Command Line Usage
    • Millifluidics Guides
      • Designing Millifluidics Using Eagle
      • Constructing Laser Cut Millifluidics
      • Constructing Millifluidics via SLA Printing
      • Calibrating IPPs
      • Operating Millifluidics
  • Troubleshooting
    • Troubleshooting Overview
    • Experiment Troubleshooting
      • Contamination
      • Vial Overflow, Pump Failure, and Spills
      • Tubing and Connector Blockage and Bursting
    • Optical Density (OD) Readings
    • Vial Troubleshooting
      • Replacing Photodiodes and LEDs
      • Heating Element
      • Stirring
    • Server Troubleshooting
    • Vial Platform Troubleshooting
      • Motherboard Troubleshooting/Replacement
      • 12V Power Supply Troubleshooting/Replacement
    • GUI Troubleshooting
    • eVOLVER Maintenance
  • Hardware
    • Overview of Hardware Architecture
    • Overview of Fluidics
      • Tubing and connectors
      • Peristaltic Pumps
      • Fluidics box
    • Overview of Millifluidics
      • IPPs (Integrated Peristaltic Pumps)
      • Pressure Regulator
    • Vial Caps
      • Universal Vial Cap Construction Guide (Luer Connectors Only)
      • 5 and 7 Port Nylon Tubing Caps Construction Protocol
    • Smart Sleeve
      • Vial Board
      • 🌪️Stirring
      • Temperature
      • Optical Density
        • OD90 vs OD135
    • Motherboard Layout and Circuitry
      • 🌡️Arduino
      • Sensor/Actuator Board Slots
      • Pulse Width Modulation (PWM) Boards
      • Analog-to-Digital Converter (ADC) Boards
    • Raspberry Pi
    • Chassis
    • Light Blocker / Splash Guard
    • Known Issues
  • Software
    • Overview of Software Architecture
    • DPU
      • Calibration
      • custom_script.py
      • Experiment Data Files
      • eVOLVER.py
    • Arduino
    • Server (Raspberry Pi)
      • Calibration Files
      • Configuration Files (conf.yml)
    • Known Issues
  • Extensions
    • Adding A New Experimental Parameter
      • Power Supply
      • Specific Applications
      • Custom Calibration Code
    • Custom Experiments
      • ePACE
        • ATTiny1634 Writing
        • LUX Board Troubleshooting
      • Morbidostat
      • Extractor Column
        • Extractor Volume Maintenance
        • Experiment Start
          • Sterilizing Extractor Fluidics
          • Setting up your experiment
          • Using the extractor script
        • Extractor Analysis
        • Troubleshooting
        • Example protocols
      • Phototroph Growth
        • Setup Phototroph eVOLVER
        • Photo-eVOLVER Smart Sleeves
          • Photo-eVOLVER Smart Sleeve Construction Guide
        • Experiment Guide
    • Custom Fluidics
      • Adding a Third Pump Rack
      • Bubblers / In-Vial Aeration
        • Bubbler Construction Protocol
        • Bubbler Cleaning Protocol
      • Running the slow pumps
    • min-eVOLVER
      • About
      • min-eVOLVER Construction
        • Parts
        • Construction Protocol
      • Fluidics Setup
      • Software Installation and Startup
      • send_command.py
      • Calibrations
      • Starting an Experiment
      • ePACE with min-eVOLVER
        • [v1.1] ePACE with min-eVOLVER
      • Troubleshooting
    • Interfacing with Other Systems
  • Contributing
    • Guidelines for Contribution
    • Reporting a Bug / Hardware Failure
    • Documentation
      • Making a Forum Post
      • How to Edit the Wiki
    • Software Development
    • Hardware Development
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  • Optogenetic Control During Continuous Culture
  • Fluorescence Measurements

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  1. Extensions
  2. Adding A New Experimental Parameter

Specific Applications

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Optogenetic Control During Continuous Culture

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.

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.

Adding pH and Dissolved Oxygen Sensors
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