Starting Experiment Checklist

1.   Prepare vials

• 24 hours before your experiment: equilibrate empty vials by opening them and storing overnight. This enables similar humidity conditions between vial and the external environment. 

2.   Prepare samples

• Mix stock solutions before transferring to test vials (vortexing is preferred, hand swirl if necessary).

3.   Check HeptaValve Mini

We recommend positioning the HeptaValve Mini body upside down for this step. 

• Verify that PEEK tubing does not have any bends or elbow as this can impact the fluidic integrity of the setup. 
• Check that the beveled end is located towards the vial or sample.Connecting the beveled end to the HeptaValve Mini inlet can cause fluidicleakage. 
• Check that the inlet numbers match the numbered labels on the PEEK tubing. 

4.   Remove translucent caps from NeOse Advance and accessories

Failing to remove caps can cause internal depression and irreversibly damage the device’s pump. 

5.   Check cable connections

• If using an external USB hub to connect accessories, ensure that you’re using the one provided by Aryballe.

• When working with the Amplifier, ensure that you’re using the plug and power cable provided by Aryballe.

6.   Set up the system

• Connect and install all the devices and accessories necessary to your experiment (NeOse Advance, HeptaValve Mini if applicable, Amplifier if applicable).

• Ensure that the HeptaValve Mini parallel to the table top after installing it on its stand. 

7.   Warm up the system 

This equilibration step allows the whole system to warm up (NeOse Advance, Amplifier, HeptaValve Mini) and ensures that the system is stable before starting your test session. 

• Use the dedicated Warm Up Protocol in the Aryballe Suite software.
• Warm up the system for at least 30 minutes prior to any measurements.

8.   Check cleanliness of your HeptaValve Mini

Launch a cleanliness check of the HeptaValve Mini without any samples connected, as detailed in the User Manual.

9.   Connect the Sample Vials

• Install a flat-ended needle in each vial, as vent system.

• Swab the flat-ended needles with a thin wire: clogging with residual silicon parts would prevent the equilibration of pressure inside the vials. 

• Using a second flat needle, pre-drill the septum for connection with the PEEK tubing.

• Connect the HeptaValve Mini to each sample vial by inserting thePEEK tubing through the septum. Make sure that the tubing does not get bent during this step.

• Double-check that vial labels, tubing numbers and experimental plan parameters entered in the Aryballe Suite setup match. 

10.Check Luer lock tightness

Loose locks affect fluidic integrity and negatively impact the intensities recorded. 

• Ensure Luer locks at the entry of the NeOse Advance and theAmplifier are tightly screwed. 

11.Calibration

If the samples have a different humidity than the ambient air, you must launch a humidity calibration: 

• The humidity levels can be assessed using the Live Data View. Make sure you present the sample for at least 30 seconds for the humidity level to stabilize.

• Use the dedicated protocol in Aryballe Suite for humidity calibration designed for your specific experimental set up: NeOse Advance alone or NeOse Advance associated to the HeptaValve Mini.

• When applying humidity calibration to your experiment, ensure that the analyte section duration for your samples is at least 40 seconds. 

12.Launch experiment and check intensities

• Using the Live Data View, check the signal intensities:

  • If the intensity is under 0.2, the signal detected is close to the limit of detection and will be mixed with the system noise. Acquisition conditions need to beadjusted so that your sample is above the detection threshold. 

  • If the signal intensity is greater than 0.2 radians, check that it is not solely linked to the sample humidity (a 10% difference of humidity levels between baseline and sample generally gives rise to a signal intensity of about 1 radian): 

    • Example 1: Mean signal intensity is 3, while Delta humidity is 25%. In this case, the signal intensity is mainly linked to the humidity of the sample (Signal/% Humidity ratio < 1).

• Example 2: Mean signal intensity is about 6, while Delta Humidity is 1.5%. In this case, the signal intensity is mainly linked to the sample (Signal/% Humidity ratio > 1).

• Using Aryballe Suite, start the acquisition protocol that is best suited to your application.
  • Observe the first cycle of the experiment to ensure proper system operation, especially: 
    • Intensity shouldr emain under 20 radians.
    • If intensity is greater than 20 radians, the sample must be diluted to prevent pollution of the system and ensure the signal detected remains in the linear range of detection.
  • Valves are opening correctly: 
    • Look at the light on the front side on the HeptaValve Mini and ensure that the LED switching position and the slight clicking sound are concurrent with the increase of signal intensity seen on the Live Data View.
    • If this is not the case, launch a Clean Up protocol. If the problem persists, please contact support@aryballe.com.
  • Valves are closing correctly
    • Observe the Live DataView: when the valve closes, the signal should descend from the plateau as shown by a slope break. Slow-moving samples can present a short delay in this break.
    • If this is not the case, launch a Clean Up protocol. If the problem persists, please contact support@aryballe.com.
  • Sensorgram returns to zero after the desorption phase:
    • If this is not the case, launch a Clean Up protocol. Increase the desorption time in subsequent experiments.
    • If possible, dilute your sample (making sure it remains over the detection threshold mentioned above) 

Summarized Checklist

The day before

   Open empty vials overnight.

On the day of the experiment

   Sample solutions are well mixed.

   HeptaValve Mini tubing is not bent.

   Beveled end of the HeptaValve Mini tubing is directed towards the vials/samples.

   HeptaValve Mini inlet numbers match the numbered labels on PEEK tubing.

   Translucent caps are removed from NeOse Advance and accessories.

   USB hub to connect accessories and Amplifier power cable are the ones provided by Aryballe.

   All devices, accessories and computer are connected.

   HeptaValve Mini body is in horizontal orientation.

   System is warmed up for at least 30 minutes. 

   Cleanliness check is performed.

   A flat-ended needle is placed in each sample vial.

   Needles are swabbed with a thin wire.

   Septa are pre-pierced with a flat-ended needle before inserting PEEK tubing through septum.

   Vial labels, tubing numbers and experimental plan parameters match.

   Luer locks on the NeOse Advance and Amplifier are tightly screwed.

   Humidity of the sample is assessed. If necessary, launch a humidity calibration.

   Samples have a signal intensity exceeding 0.2 and a signal intensity/ %humidity ratio greater than 1. 

   Intensity is lower than 20 radians.

   Valves are opening correctly.

   Valves are closing correctly. 

   Sensorgram returns to zero after desorption.