NT8 User's Guide | Chapter 10: Hanging Drop Experiments
● Verify that columns 1-12 are selected next to Column (if selected, they appear in
dark blue). If not, click column 1, then press and hold the SHIFT key and click
column 12 to select all columns.
● Next to Volume, enter the desired dispense volume (generally the combined
volume of the well solution and the protein volume).
● Optional: Next to Mixing, select the number of mixing cycles you want the NT8
to perform before dispensing drops.
● Optional: Next to Z-Offset, you may choose to leave Flexible Fingers enabled
(indicated by the FF symbol), or you may click the FF button and then manually
enter the distance the pipette tips should travel before dispensing your drops. It is
strongly recommended that you leave Flexible Fingers enabled for this type of
dispense.
● Select a Liquid Class from the liquid class list.
● Click Add.
8. Click End Repeat in the Task List to select it.
9. Double-click the Waste Station to add the Dispose Tip command to the Task List.
10. Optional: Save your Task List by clicking the Save button .
Add Protein to the Protein Block
● Using a hand pipette, pipette the protein to be used in the hanging drop experiment
into the protein block.
Run the Hanging Drop Dispense
● Click the Run Task List button to execute the Task List. The NT8 head moves to
the LV Tip Caddy, picks up a column of tips, then washes them at the wash station.
Next, the NT8 will aspirate protein from the protein block, then aspirate solution from
the microplate well. Finally, the NT8 will dispense the hanging drops to the UVP
Hanging Drop Plate Seal. This series of actions will be repeated on each column
specified. Finally, the NT8 will dispose the used tips into the waste station.
Transfer the Hanging Drops to the Microplate
1. When the dispense has completed, take the microplate and place it on the HD seal jig
transfer. The A1 position should be at the top-left corner.
2. Place the UVP Seal and the reference plate on the microplate by carefully flipping it
over horizontally above the microplate. Use the three steel pins to guide the reference
plate into place. The angled edges of the UVP Seal should match the angled edges of
the microplate. Once in place, press down firmly.
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