Sitting Drop Experiments with 24 Well Plate Type¶
Starting from software version 4.4, NT8 supports the use of 24-well plate type to run Sitting Drop experiments. Follow this tutorial to set up the 24-well plate in your NT8.
Prerequisites¶
You have upgraded the NT8 hardware to the redesigned waste station design.
The Redesigned Waste Station
You are using the NT8 Control Software version 4.4 or newer.
The 24-well plate type feature has been enabled in the Device.config. Contact support@formulatrix.com. for assistance.
Power On the NT8 Hardware and Start the Control Software¶
Connect the NT8 to your computer using the USB cable.
Find the power switch located on the back-right of the NT8 and turn it on.
Double-click the NT8 icon on your computer’s desktop. It will take a few moments for the NT8 Control Software to load while the NT8 initializes. The instrument indicator on the bottom-left corner of the software will turn green when the NT8 is connected and ready to use.
After that, proceed to Set Up the NT8 Hardware.
Set Up the NT8 Hardware¶
Place the LV tip caddy and the 24-well Sitting Drop Experiment plate on the NT8.
Place the protein tray on the protein block, and put the protein tray clamp on top of the protein tray to hold it in place.
Note
You should add the protein to the Protein Tray after you have set up the task list in the software. Make sure the protein samples are stored in wells number 2, 4, 6, and 8 of the Protein Tray.
Option 1: Set Up the NT8 Control Software to Work with Sitting Drop Template
The Sitting Drop setup template with 24-well plate types is pre-installed with your software. You may use it if you plan to combine up to 750 nL protein and 750 nL well solution for a 1500 nL dispense. You can modify these volumes by double-clicking the item on the task list. The Hampton Cryschem Plate is the default plate for the Sitting Drop template.
Note
You can add other types of 24-well Sitting Drop Experiment plates from the Plate Type Editor options from the Tools menu.
To use this template:
Click the Open Task List button
in the Task List panel.OR
Click the File menu and select Load Task List.
Open the Sample folder.
Select Hampton Cryschem S Plate 750 nL Protein and 750 nL Well Solution task list.
Note
The maximum volume capacity of LV Tips is 1500 nL. If you want to dispense more than 1500 nL, please add the aspirate/dispense task twice.
Click Open.
OPTIONAL: If you need to change the aspirate or dispense volumes, double-click the task in the Task List panel and edit the volume.
Option 2: Manually Set Up the NT8 Control Software for Sitting Drop Experiments with 24-well Plate Type
Mimic the NT8’s physical setup with the software. Under the associated plate station on the software, click the downward arrow and select the 24 1 Hampton CryschemSPlate plate from the Plate Selection window.
Click the Add Repeat Task button
on the Task List to start the beginning of tasks that will be repeated on every column of a plate.Double-click the LV Tip Caddy image to add the Pick Tip command to the Task List.
Double-click the Wash Station image to add the Wash Tip command to the task list. The default Wash Time is two seconds. You can adjust it by double-clicking the Wash Tip action in the Task List panel.
Tell the NT8 Control Software to aspirate protein from the protein block:
Double-click the Protein Block image to open the Task Details window.
Task Details Window for Protein Aspiration
Under Select Operation, select Aspirate. The Low Volume Tips under Select Head option is automatically selected.
Under Column, select the column where you’ll aspirate the protein from.
Select a Liquid Class from the liquid class list.
Next to Volume, type in the volume you want to aspirate.
OPTIONAL: Next to the Mixing Cycle, enter the number of times you want the volume to be mixed.
OPTIONAL: Next to Mixing Volume, set the volume in nL to be mixed.
OPTIONAL: Set the additional volume in nL to be aspirated after aspirating the targeted volume in the Pre-Dispense box.
OPTIONAL: You may opt for Custom Aspirate to determine the tip height during an aspiration task. However, we recommend that you use Smart Aspirate so that the tip height will automatically adjust to the remaining volume in the protein tray. Smart Aspirate is enabled by default.
If you are finished adding commands for that plate, click Add. If you want to create another aspirate task, click Add More.
Now you’ll tell the NT8 to aspirate your well solution.
Double-click the microplate image to open the Task Details window.
Task Details Window for Well Solution Aspiration
Under Select Operation, click Aspirate. The Low Volume Tips in the Select Head option is automatically selected.
Under Location drop-down list, select Well. The selected location is highlighted in yellow.
Selecting Target Area
To select multiple columns, click the first desired column, then hold the Shift key and click the last desired column.
Select a Liquid Class from the liquid class list.
Set the volume to be aspirated in the Volume box.
OPTIONAL: Set the additional volume to be aspirated after aspirating the targeted volume in the Pre-Dispense box.
Next to Z-Offset, determine the height in mm at which the tip stops before aspirating.
If you are finished adding commands for that plate, click Add. If you want to dispense to drop locations on this plate, click Add More.
On your Task List panel, the Prepare Tips task is automatically added before protein aspiration. This task discards 4 tips before aspirating the protein to make it compatible with the 24-well plate dispense.
Instruct the NT8 to dispense the protein and well solution mixture to the drop location on the microplate.
If you have clicked Add More in the last step, the Task Details window remains open. If not, double-click the microplate image again to open it.
Under Select Operation, click Dispense.
Under Location drop-down list, select Drop 1. The selected location is highlighted in yellow.
Selecting Target Area
Next to Column, select the columns you want to dispense to.
Select a Liquid Class from the liquid class list.
Next to Volume, enter the volume to be dispensed. If you want the NT8 to mix your solution, type the number of mixing cycles next to Mixing (optional).
Next to Z-Offset, you may choose to let the NT8’s Flexible Fingers determine the dispense height (recommended), or manually specify the distance you want the fingers to stop at before dispensing.
If you are finished adding commands for that plate, click Add. If you want to create more dispense tasks to this plate, click Add More.
Task Details Window for Protein and Well Solution Dispensing
Add the Dispose Tip command to the Task List by clicking the Waste Station image. In the Add Task window, select Dispose all tips.
Note
For each column, the hardware will pick up new tips and discard all of them at the end of each column dispense.
Disposing All Tips after Dispensing
OPTIONAL: Save your Task List by clicking the Save Task List button .
OR
Click the File menu and select Save Task List. Your task list will look like this.
24-well Plate Sitting Drop Task List
Add Protein to the Protein Block for 24-well Plate Dispensing¶
Prerequisite:
You have prepared a hand pipette.
Important
You only need to fill the protein sample in the selected wells (the wells number 2, 4, 6, and 8) of the Protein Tray (indicated by the blue color below).
You Must Fill the Protein into the Wells Indicated in Blue Only
To fill the protein sample, pipette the protein sample to be used in a Sitting Drop experiment to the protein tray.
Run the 24-well Plate Dispensing Task List¶
Click the Run Task List button 
to execute the Task List. The NT8 picks up the LV tips, washes them, prepares 4 tips, aspirates from the protein block, aspirates from the wells in the microplate, and then dispenses to the drop location. This process repeats for each column you specified earlier. After the NT8 has completed the tasks for each column, it disposes all tips in the waste station.
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