week-03-hw-lab-automation

🤖 Opentrons Liquid-Handling Artwork

🧠 Project Overview

This project transforms the Opentrons OT-2 liquid handling robot into a biological plotter.
Using coordinate-based programming, the robot deposits fluorescent bacterial droplets onto an agar plate to form a structured artistic pattern.

The objective was to:

Convert digital coordinates into physical bacterial deposition
Control droplet detachment to avoid agar smearing
Implement automated refill logic
Validate the protocol using simulation before execution

🎨 Artistic Concept – Yin Yang Design

The selected design is inspired by the Yin-Yang symbol, representing:

Balance between automation and biology
Precision vs. organic growth
Engineering control vs. living systems

Final Design

🧪 Experimental Configuration

Robot: Opentrons OT-2
Pipette: P20 Single Channel Gen2
Agar Plate: Custom HTGAA agar plate
Fluorescent Strains:

Color	Fluorescent Marker
Green	mClover3
Orange	Azurite

Each coordinate corresponds to a 1.5 µL droplet deposited at a specific XY location relative to the plate center.

🧾 Full Simulation Python Script

Below is the complete script used for the Opentrons simulation and execution.

from opentrons import types

import string

metadata = {
    'protocolName': 'Juan Francisco Larrea - Opentrons Art - HTGAA',
    'author': 'HTGAA',
    'source': 'HTGAA 2026',
    'apiLevel': '2.20'
}

Z_VALUE_AGAR = 2.0
POINT_SIZE = 1.5

mclover3_points = [(-11.25,31.25), (-8.75,31.25), (-6.25,31.25), (-3.75,31.25), (-1.25,31.25), (1.25,31.25), (3.75,31.25), (6.25,31.25), (8.75,31.25), (-16.25,28.75), (-13.75,28.75), (-11.25,28.75), (8.75,28.75), (11.25,28.75), (13.75,28.75), (-18.75,26.25), (-16.25,26.25), (13.75,26.25), (16.25,26.25), (18.75,26.25), (-21.25,23.75), (13.75,23.75), (16.25,23.75), (18.75,23.75), (21.25,23.75), (-23.75,21.25), (16.25,21.25), (18.75,21.25), (21.25,21.25), (23.75,21.25), (-26.25,18.75), (-1.25,18.75), (1.25,18.75), (18.75,18.75), (21.25,18.75), (23.75,18.75), (26.25,18.75), (-26.25,16.25), (-3.75,16.25), (-1.25,16.25), (1.25,16.25), (3.75,16.25), (18.75,16.25), (21.25,16.25), (23.75,16.25), (26.25,16.25), (-28.75,13.75), (-3.75,13.75), (-1.25,13.75), (1.25,13.75), (3.75,13.75), (18.75,13.75), (21.25,13.75), (23.75,13.75), (26.25,13.75), (28.75,13.75), (-28.75,11.25), (-1.25,11.25), (1.25,11.25), (16.25,11.25), (18.75,11.25), (21.25,11.25), (23.75,11.25), (26.25,11.25), (28.75,11.25), (-28.75,8.75), (13.75,8.75), (16.25,8.75), (18.75,8.75), (21.25,8.75), (23.75,8.75), (26.25,8.75), (28.75,8.75), (-31.25,6.25), (13.75,6.25), (16.25,6.25), (18.75,6.25), (21.25,6.25), (23.75,6.25), (26.25,6.25), (28.75,6.25), (31.25,6.25), (-31.25,3.75), (11.25,3.75), (13.75,3.75), (16.25,3.75), (18.75,3.75), (21.25,3.75), (23.75,3.75), (26.25,3.75), (28.75,3.75), (31.25,3.75), (-31.25,1.25), (6.25,1.25), (8.75,1.25), (11.25,1.25), (13.75,1.25), (16.25,1.25), (18.75,1.25), (21.25,1.25), (23.75,1.25), (26.25,1.25), (28.75,1.25), (31.25,1.25), (-31.25,-1.25), (-3.75,-1.25), (-1.25,-1.25), (1.25,-1.25), (3.75,-1.25), (6.25,-1.25), (8.75,-1.25), (11.25,-1.25), (13.75,-1.25), (16.25,-1.25), (18.75,-1.25), (21.25,-1.25), (23.75,-1.25), (26.25,-1.25), (28.75,-1.25), (31.25,-1.25), (-31.25,-3.75), (-8.75,-3.75), (-6.25,-3.75), (-3.75,-3.75), (-1.25,-3.75), (1.25,-3.75), (3.75,-3.75), (6.25,-3.75), (8.75,-3.75), (11.25,-3.75), (13.75,-3.75), (16.25,-3.75), (18.75,-3.75), (21.25,-3.75), (23.75,-3.75), (26.25,-3.75), (28.75,-3.75), (31.25,-3.75), (-31.25,-6.25), (-11.25,-6.25), (-8.75,-6.25), (-6.25,-6.25), (-3.75,-6.25), (-1.25,-6.25), (1.25,-6.25), (3.75,-6.25), (6.25,-6.25), (8.75,-6.25), (11.25,-6.25), (13.75,-6.25), (16.25,-6.25), (18.75,-6.25), (21.25,-6.25), (23.75,-6.25), (26.25,-6.25), (28.75,-6.25), (31.25,-6.25), (-28.75,-8.75), (-11.25,-8.75), (-8.75,-8.75), (-6.25,-8.75), (-3.75,-8.75), (-1.25,-8.75), (1.25,-8.75), (3.75,-8.75), (6.25,-8.75), (8.75,-8.75), (11.25,-8.75), (13.75,-8.75), (16.25,-8.75), (18.75,-8.75), (21.25,-8.75), (23.75,-8.75), (26.25,-8.75), (28.75,-8.75), (-28.75,-11.25), (-13.75,-11.25), (-11.25,-11.25), (-8.75,-11.25), (-6.25,-11.25), (-3.75,-11.25), (-1.25,-11.25), (1.25,-11.25), (3.75,-11.25), (6.25,-11.25), (8.75,-11.25), (11.25,-11.25), (13.75,-11.25), (16.25,-11.25), (18.75,-11.25), (21.25,-11.25), (23.75,-11.25), (26.25,-11.25), (28.75,-11.25), (-28.75,-13.75), (-13.75,-13.75), (-11.25,-13.75), (-8.75,-13.75), (-6.25,-13.75), (-3.75,-13.75), (3.75,-13.75), (6.25,-13.75), (8.75,-13.75), (11.25,-13.75), (13.75,-13.75), (16.25,-13.75), (18.75,-13.75), (21.25,-13.75), (23.75,-13.75), (26.25,-13.75), (28.75,-13.75), (-26.25,-16.25), (-13.75,-16.25), (-11.25,-16.25), (-8.75,-16.25), (-6.25,-16.25), (6.25,-16.25), (8.75,-16.25), (11.25,-16.25), (13.75,-16.25), (16.25,-16.25), (18.75,-16.25), (21.25,-16.25), (23.75,-16.25), (26.25,-16.25), (-26.25,-18.75), (-13.75,-18.75), (-11.25,-18.75), (-8.75,-18.75), (-6.25,-18.75), (6.25,-18.75), (8.75,-18.75), (11.25,-18.75), (13.75,-18.75), (16.25,-18.75), (18.75,-18.75), (21.25,-18.75), (23.75,-18.75), (26.25,-18.75), (-23.75,-21.25), (-13.75,-21.25), (-11.25,-21.25), (-8.75,-21.25), (-6.25,-21.25), (-3.75,-21.25), (3.75,-21.25), (6.25,-21.25), (8.75,-21.25), (11.25,-21.25), (13.75,-21.25), (16.25,-21.25), (18.75,-21.25), (21.25,-21.25), (23.75,-21.25), (-21.25,-23.75), (-11.25,-23.75), (-8.75,-23.75), (-6.25,-23.75), (-3.75,-23.75), (-1.25,-23.75), (1.25,-23.75), (3.75,-23.75), (6.25,-23.75), (8.75,-23.75), (11.25,-23.75), (13.75,-23.75), (16.25,-23.75), (18.75,-23.75), (21.25,-23.75), (-18.75,-26.25), (-16.25,-26.25), (-11.25,-26.25), (-8.75,-26.25), (-6.25,-26.25), (-3.75,-26.25), (-1.25,-26.25), (1.25,-26.25), (3.75,-26.25), (6.25,-26.25), (8.75,-26.25), (11.25,-26.25), (13.75,-26.25), (16.25,-26.25), (18.75,-26.25), (-13.75,-28.75), (-11.25,-28.75), (-8.75,-28.75), (-6.25,-28.75), (-3.75,-28.75), (-1.25,-28.75), (1.25,-28.75), (3.75,-28.75), (6.25,-28.75), (8.75,-28.75), (11.25,-28.75), (13.75,-28.75), (-11.25,-31.25), (-8.75,-31.25), (-6.25,-31.25), (-3.75,-31.25), (-1.25,-31.25), (1.25,-31.25), (3.75,-31.25), (6.25,-31.25), (8.75,-31.25)]
azurite_points = [(-8.75,28.75), (-6.25,28.75), (-3.75,28.75), (-1.25,28.75), (1.25,28.75), (3.75,28.75), (6.25,28.75), (-13.75,26.25), (-11.25,26.25), (-8.75,26.25), (-6.25,26.25), (-3.75,26.25), (-1.25,26.25), (1.25,26.25), (3.75,26.25), (6.25,26.25), (8.75,26.25), (11.25,26.25), (-18.75,23.75), (-16.25,23.75), (-13.75,23.75), (-11.25,23.75), (-8.75,23.75), (-6.25,23.75), (-3.75,23.75), (-1.25,23.75), (1.25,23.75), (3.75,23.75), (6.25,23.75), (8.75,23.75), (11.25,23.75), (-21.25,21.25), (-18.75,21.25), (-16.25,21.25), (-13.75,21.25), (-11.25,21.25), (-8.75,21.25), (-6.25,21.25), (-3.75,21.25), (-1.25,21.25), (1.25,21.25), (3.75,21.25), (6.25,21.25), (8.75,21.25), (11.25,21.25), (13.75,21.25), (-23.75,18.75), (-21.25,18.75), (-18.75,18.75), (-16.25,18.75), (-13.75,18.75), (-11.25,18.75), (-8.75,18.75), (-6.25,18.75), (-3.75,18.75), (3.75,18.75), (6.25,18.75), (8.75,18.75), (11.25,18.75), (13.75,18.75), (16.25,18.75), (-23.75,16.25), (-21.25,16.25), (-18.75,16.25), (-16.25,16.25), (-13.75,16.25), (-11.25,16.25), (-8.75,16.25), (-6.25,16.25), (6.25,16.25), (8.75,16.25), (11.25,16.25), (13.75,16.25), (16.25,16.25), (-26.25,13.75), (-23.75,13.75), (-21.25,13.75), (-18.75,13.75), (-16.25,13.75), (-13.75,13.75), (-11.25,13.75), (-8.75,13.75), (-6.25,13.75), (6.25,13.75), (8.75,13.75), (11.25,13.75), (13.75,13.75), (16.25,13.75), (-26.25,11.25), (-23.75,11.25), (-21.25,11.25), (-18.75,11.25), (-16.25,11.25), (-13.75,11.25), (-11.25,11.25), (-8.75,11.25), (-6.25,11.25), (-3.75,11.25), (3.75,11.25), (6.25,11.25), (8.75,11.25), (11.25,11.25), (13.75,11.25), (-26.25,8.75), (-23.75,8.75), (-21.25,8.75), (-18.75,8.75), (-16.25,8.75), (-13.75,8.75), (-11.25,8.75), (-8.75,8.75), (-6.25,8.75), (-3.75,8.75), (-1.25,8.75), (1.25,8.75), (3.75,8.75), (6.25,8.75), (8.75,8.75), (11.25,8.75), (-28.75,6.25), (-26.25,6.25), (-23.75,6.25), (-21.25,6.25), (-18.75,6.25), (-16.25,6.25), (-13.75,6.25), (-11.25,6.25), (-8.75,6.25), (-6.25,6.25), (-3.75,6.25), (-1.25,6.25), (1.25,6.25), (3.75,6.25), (6.25,6.25), (8.75,6.25), (11.25,6.25), (-28.75,3.75), (-26.25,3.75), (-23.75,3.75), (-21.25,3.75), (-18.75,3.75), (-16.25,3.75), (-13.75,3.75), (-11.25,3.75), (-8.75,3.75), (-6.25,3.75), (-3.75,3.75), (-1.25,3.75), (1.25,3.75), (3.75,3.75), (6.25,3.75), (8.75,3.75), (-28.75,1.25), (-26.25,1.25), (-23.75,1.25), (-21.25,1.25), (-18.75,1.25), (-16.25,1.25), (-13.75,1.25), (-11.25,1.25), (-8.75,1.25), (-6.25,1.25), (-3.75,1.25), (-1.25,1.25), (1.25,1.25), (3.75,1.25), (-28.75,-1.25), (-26.25,-1.25), (-23.75,-1.25), (-21.25,-1.25), (-18.75,-1.25), (-16.25,-1.25), (-13.75,-1.25), (-11.25,-1.25), (-8.75,-1.25), (-6.25,-1.25), (-28.75,-3.75), (-26.25,-3.75), (-23.75,-3.75), (-21.25,-3.75), (-18.75,-3.75), (-16.25,-3.75), (-13.75,-3.75), (-11.25,-3.75), (-28.75,-6.25), (-26.25,-6.25), (-23.75,-6.25), (-21.25,-6.25), (-18.75,-6.25), (-16.25,-6.25), (-13.75,-6.25), (-26.25,-8.75), (-23.75,-8.75), (-21.25,-8.75), (-18.75,-8.75), (-16.25,-8.75), (-13.75,-8.75), (-26.25,-11.25), (-23.75,-11.25), (-21.25,-11.25), (-18.75,-11.25), (-16.25,-11.25), (-26.25,-13.75), (-23.75,-13.75), (-21.25,-13.75), (-18.75,-13.75), (-16.25,-13.75), (-1.25,-13.75), (1.25,-13.75), (-23.75,-16.25), (-21.25,-16.25), (-18.75,-16.25), (-16.25,-16.25), (-3.75,-16.25), (-1.25,-16.25), (1.25,-16.25), (3.75,-16.25), (-23.75,-18.75), (-21.25,-18.75), (-18.75,-18.75), (-16.25,-18.75), (-3.75,-18.75), (-1.25,-18.75), (1.25,-18.75), (3.75,-18.75), (-21.25,-21.25), (-18.75,-21.25), (-16.25,-21.25), (-1.25,-21.25), (1.25,-21.25), (-18.75,-23.75), (-16.25,-23.75), (-13.75,-23.75), (-13.75,-26.25)]

point_name_pairing = [("Green", mclover3_points),("Orange", azurite_points)]

# Robot deck setup constants
TIP_RACK_DECK_SLOT = 9
COLORS_DECK_SLOT = 6
AGAR_DECK_SLOT = 5
PIPETTE_STARTING_TIP_WELL = 'A1'

# Place the PCR tubes in this order
well_colors = {
    'A1' : 'Red',
    'B1' : 'Green',
    'C1' : 'Orange'
}

# Initialize volume_used globally
volume_used = {}

def update_volume_remaining(current_color, quantity_to_aspirate):
    global well_colors
    global volume_used

    rows = string.ascii_uppercase
    cols_str = [str(i) for i in range(1, 13)] # Columns 1 to 12

    if current_color not in volume_used:
        volume_used[current_color] = 0

    # Find the current well for this color
    current_well_for_color = None
    for well, color in list(well_colors.items()):
        if color == current_color:
            current_well_for_color = well
            break

    if current_well_for_color is None:
        raise ValueError(f"Color {current_color} not found in well_colors for volume update.")

    if (volume_used[current_color] + quantity_to_aspirate) > 250:
        row_letter = current_well_for_color[0]
        col_number_str = current_well_for_color[1:]

        next_col_index = cols_str.index(col_number_str) + 1
        if next_col_index >= len(cols_str):
            raise IndexError(f"Ran out of wells for color {current_color} in row {row_letter} (max column reached)!")

        next_well = f"{row_letter}{cols_str[next_col_index]}"

        # Remove the old well from well_colors map, and add the new one.
        # This is safe because each color is assumed to have its own row.
        del well_colors[current_well_for_color]
        well_colors[next_well] = current_color
        volume_used[current_color] = quantity_to_aspirate # Reset volume for new well
    else:
        volume_used[current_color] += quantity_to_aspirate

def run(protocol):
  ##############################################################################
  ###   Load labware, modules and pipettes
  ##############################################################################

  # Tips
  tips_20ul = protocol.load_labware('opentrons_96_tiprack_20ul', TIP_RACK_DECK_SLOT, 'Opentrons 20uL Tips')

  # Pipettes
  pipette_20ul = protocol.load_instrument("p20_single_gen2", "right", [tips_20ul])

  # Modules
  temperature_module = protocol.load_module('temperature module gen2', COLORS_DECK_SLOT)

  # Temperature Module Plate
  temperature_plate = temperature_module.load_labware('opentrons_96_aluminumblock_generic_pcr_strip_200ul',
                                                    'Cold Plate')
  # Choose where to take the colors from
  color_plate = temperature_plate

  # Agar Plate
  agar_plate = protocol.load_labware('htgaa_agar_plate', AGAR_DECK_SLOT, 'Agar Plate')  ## TA MUST CALIBRATE EACH PLATE!
  # Get the top-center of the plate, make sure the plate was calibrated before running this
  center_location = agar_plate['A1'].top()

  pipette_20ul.starting_tip = tips_20ul.well(PIPETTE_STARTING_TIP_WELL)


  # Helper function (color location)
  def location_of_color(color_string):
    for well,color in well_colors.items():
      if color.lower() == color_string.lower():
        return temperature_plate[well]
    raise ValueError(f"No well found with color {color_string}")

  # For this lab, instead of calling pipette.dispense(1, loc) use this: dispense_and_detach(pipette, 1, loc)
  def dispense_and_detach(pipette, volume, location):
      """
      Move laterally 5mm above the plate (to avoid smearing a drop); then drop down to the plate,
      dispense, move back up 5mm to detach drop, and stay high to be ready for next lateral move.
      5mm because a 4uL drop is 2mm diameter; and a 2deg tilt in the agar pour is >3mm difference across a plate.
      """
      assert(isinstance(volume, (int, float)))
      above_location = location.move(types.Point(z=location.point.z + 5))  # 5mm above
      pipette.move_to(above_location)       # Go to 5mm above the dispensing location
      pipette.dispense(volume, location)    # Go straight downwards and dispense
      pipette.move_to(above_location)       # Go straight up to detach drop and stay high



  # Print pattern by iterating over lists
  for idx, (current_color, point_list) in enumerate(point_name_pairing): # Renamed i to idx to avoid conflict
      # Skip the rest of the loop if the list is empty
      if not point_list:
          continue

      pipette_20ul.pick_up_tip()

      max_aspirate = int(18 // POINT_SIZE) * POINT_SIZE
      quantity_to_aspirate = min(len(point_list)*POINT_SIZE, max_aspirate)

      # Get the initial well for this color before any volume updates
      initial_aspirate_well = location_of_color(current_color)

      # Update volume (this might change `well_colors` for `current_color`)
      update_volume_remaining(current_color, quantity_to_aspirate)

      # Aspirate from the (potentially updated) location
      pipette_20ul.aspirate(quantity_to_aspirate, location_of_color(current_color))

      # Iterate over the current points list and dispense them, refilling along the way
      for j in range(len(point_list)):
          x, y = point_list[j]
          adjusted_location = center_location.move(types.Point(x, y))

          dispense_and_detach(pipette_20ul, POINT_SIZE, adjusted_location)

          if pipette_20ul.current_volume == 0 and len(point_list[j+1:]) > 0:
              # Need to refill
              refill_quantity = min(len(point_list[j+1:])*POINT_SIZE, max_aspirate)

              # Get the current source well for this color *before* updating volume, in case it changes
              previous_refill_well = location_of_color(current_color)

              # Update volume and potentially move the color to a new physical well
              update_volume_remaining(current_color, refill_quantity)

              # Get the (potentially new) source well for this color
              new_refill_well = location_of_color(current_color)

              if new_refill_well != previous_refill_well:
                  # If the source well has changed for this color, we must drop the tip and pick up a new one
                  pipette_20ul.drop_tip()
                  pipette_20ul.pick_up_tip()

              # Now aspirate from the correct (potentially new) well
              pipette_20ul.aspirate(refill_quantity, new_refill_well)

      # Drop tip between each color
      pipette_20ul.drop_tip()

🖥️ Simulation Output

Before running on the real robot, the protocol was validated using the Opentrons simulator.

📊 Result

The robot successfully deposited bacteria following the coordinate map. After incubation, bacterial growth revealed the intended image on the agar plate.

This demonstrates that liquid-handling robots can perform microscale spatial biofabrication, a technique related to:

tissue engineering
biosensors
living materials

Post-Lab Questions

1. Find and describe a published paper that utilizes the Opentrons or an automation tool to achieve novel biological applications.

🧪 HYDRA: Automated Hydrogel Fabrication for High-Throughput Drug Screening

Figure. Overview of the HYDRA method.
A liquid-handling robot dispenses and re-aspirates hydrogel precursor solution to leave a micrometer-thin planar hydrogel inside multi-well plates, enabling biomimetic cell culture compatible with high-throughput drug screening and imaging.

HYDRA (HYDrogels by Robotic liquid handling Automation) presents a scalable and automated method to fabricate thin, uniform hydrogel layers inside standard multi-well plates used for high-throughput screening (HTS).

Traditional cell culture relies on rigid plastic or glass substrates, which poorly mimic the mechanical environment of real human tissues. This lack of physiological relevance contributes to high drug failure rates in clinical trials.
HYDRA addresses this issue by introducing planar hydrogel coatings (10–50 µm thick) that better replicate tissue stiffness while remaining fully compatible with imaging-based screening systems.

The key challenge solved by the study is meniscus formation in small wells, which normally leads to uneven gel surfaces and poor imaging quality. The authors developed a robotic workflow that deposits and re-aspirates hydrogel precursor solution to leave behind a controlled, micrometer-thin film.

The system was validated using:

Epithelial cell culture (HaCaT cells)
Dose-response experiments with anticancer drugs (nocodazole, paclitaxel)
Digital holography and fluorescence microscopy

Results demonstrated:

Reproducible gel thickness
High imaging compatibility
Scalability to 96- and 384-well formats

Conclusion: HYDRA enables more biomimetic and predictive drug testing without requiring new laboratory infrastructure.

🤖 How HYDRA Uses Opentrons & Automation

The innovation of the paper lies in combining biomaterials + robotics.

An Opentrons OT-2 liquid-handling robot was programmed to:

⚙️ Automated Workflow

Mix fish gelatin and transglutaminase precursor solutions.
Dispense a small volume at the center of each well.
Avoid touching the sidewalls (to prevent meniscus formation).
Immediately re-aspirate the same volume.
Leave behind a thin liquid boundary layer.
Allow enzymatic crosslinking to form a flat hydrogel film. The protocol was implemented using:

Opentrons Protocol Designer
Custom Python scripts
Calibrated pipette heights and flow rates
Precise aspiration control

🚀 Why Automation Matters

Using Opentrons transforms hydrogel fabrication into a standardized, scalable microfabrication process:

✅ High reproducibility
✅ Compatible with existing HTS pipelines
✅ Rapid fabrication (~10 minutes per plate)
✅ No specialized hardware required

Instead of using the robot as a simple pipetting tool, HYDRA turns it into a biomaterial fabrication platform — enabling physiologically relevant substrates directly inside standard drug screening plates.

2. Write a description about what you intend to do with automation tools for your final project. You may include example pseudocode, Python scripts, 3D printed holders, a plan for how to use Ginkgo Nebula, and more.

This project proposes to automate the screening and characterization of nanoparticle delivery systems for the delivery of the damage suppressor protein (Dsup) — a nucleosome-binding protein from Ramazzottius varieornatus (tardigrade). Dsup has been demonstrated to protect mammalian cells from oxidative stress and UV-induced DNA damage.

The goal is to determine which nanoparticle formulation most effectively delivers Dsup into human dermal fibroblasts, improving resistance to oxidative stress (H₂O₂ exposure). The long-term application is skin regeneration and anti-aging therapies.

Automation tools (Opentrons OT-2 + cloud lab integration) will be used to:

Prepare nanoparticle formulations
Perform controlled protein loading
Treat fibroblast cultures
Apply oxidative stress
Perform viability assays
Collect quantitative data

⚙️ Automated Workflow

flowchart TD
    START[Start Protocol]
    START --> LOAD[Load Labware & Tips]
    LOAD --> PREP_NP[Prepare Nanoparticles]
    PREP_NP --> LOAD_DSUP[Load Dsup Protein]
    LOAD_DSUP --> INCUBATE1[Incubate Protein Loading]
    INCUBATE1 --> SEED[Seed Fibroblasts]
    SEED --> ADD_TREATMENT[Add NP-Dsup Treatment]
    ADD_TREATMENT --> INCUBATE2[24h Incubation]
    INCUBATE2 --> ADD_STRESS[Add H2O2]
    ADD_STRESS --> INCUBATE3[Stress Incubation]
    INCUBATE3 --> ADD_ASSAY[Add Viability Reagent]
    ADD_ASSAY --> READ[Transfer to Reading Plate]
    READ --> END[Export Data]

☁️ Cloud laboratory for large-scale validation:

Instrument	Function
Echo	Transfers precise nanoliter volumes of Dsup protein and reagents
Bravo	Dispenses cell-free protein expression (CFPS) reagents into plates
Multiflo	Adds media, buffers, and treatment solutions across wells
Inheco	Provides controlled temperature incubation during reactions
PlateLoc	Seals microplates to prevent contamination and evaporation
PHERAstar	Measures fluorescence output for cell viability and protein activity