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FluidFM Probe Coating with PAcrAm

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Maria Milla


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In this protocol, the FluidFM probe is coated with PAcrAm (SuSoS AG). This compound is a polymer used as antifouling agent for glass and polymer cell culture substrates. This coating is ideal for coating micropipettes in isolation workflows, but it also can be used for nanosyringe coating for the injection of specific compounds. 

PAcrAm is distributed in 0.1 mg lyophilized aliquots, which also contain the buffer required for surface modification.  

There are two different protocols for probe coating using PAcrAm, depending if they are coated during the experiment or before.

a. COATING WITH PAcrAm inside FluidFM system

This is ideal if you want the coating to be very fresh before starting an experiment. Only 1 probe can be coated like this at a time.

1. If stored at -20°C, temperate the PAcrAm aliquot at room temperature. Centrifuge the aliquot for 5 s and dilute the content in 1 ml of ultra-pure water. Vortex for 5 s.

2. In a 12-well plate (ThermoFisher, Nunc), dispense all the PAcrAm solution into the position A1. Dispense also 1 ml of PBS in A2, A3 and A4. 

3. Use the Exchange Plates workflow in ARYA to place the coating plate on the left port. Place the probe loading plate in the right port of the BOT by using the Exchange Plates workflow. 

4. Fill the reservoir of the FluidFM probe with 1 ul of your solution (e.g. for isolation workflow, use Trypsin 10x). 

5. Place the probe in the probe loading plate. 

6. Select Preparation workflow and follow it until the step Go to Sample. In this step, right after filling the probe, dip the FluidFM probe into the PAcrAm solution on the plate placed in the left port (A1 position), using the Navigation Tool. 

7. Incubate the probe into PAcrAm solution for 30 min.

8. Wash 3 times in PBS by moving into the 3 wells containing PBS (A2, A3, A4). The probe is now ready to use.

b. Pre-COATING Probes WITH PAcrAm

This protocol allows you to coat multiple probes the day before the experiment, allowing shorter setup time and higher throughput during the experiment.

1. If stored at -20°C, temperate the PAcrAm aliquot at room temperature. Centrifuge the aliquot for 5 s and dilute the content in 1 ml of ultra-pure water. Vortex for 5 s. 

2. In a 12-well plate (ThermoFisher, Nunc), dispense all the PAcrAm solution into the position A1. Dispense also 1 ml of ultra-pure water in A2, A3 and A4. 

3. Place the FluidFM storage plate on top of the 12-well plate. 

4. Place the probe to be coated on top of the well containing the PAcrAm, making sure that the probe is in contact with the PAcrAm solution. 

5. Incubate for 30 min at room-temperature.   

5. Wash 3 times in ultra-pure water by dipping the probe for 5 sec in the other 3 wells (A2, A3, A4). 

6. Place the probe into a desiccator and apply vacuum over-night to dry the probe. The probe is now ready to be used. Be aware that the coating is not stable over time. Therefore, we recommend using the coated probe in the next 2 days after the coating.


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