In this tutorial, I will show you how to create a simple protocol that contains a hit-picking routine, a serial dilution routine, and a replication routine using the VWorks software.
The tutorial consists of five short videos: Introduction, Hit Picking, Serial Dilution, Plate Replication, Compile Protocol and Simulate Runs.
This tutorial assumes that you already know how to create a device file in the VWorks software. If you do not have this background, please view the Device File video first before
continuing. You can also learn about device files and writing basic protocols in the VWorks Automation Control User Guide.
Before I start creating the protocol in the VWorks software, I like to outline what my protocol will do. I have six source plates. I want to use the protocol to pick two or three samples
from each source plate (a total of 16 samples), dispense the samples in the first column in the 384-well destination plate, dilute the column serially in the destination plate,
and then replicate the destination plate twice.
I have to let the software know which source wells I want to pick from. For this, I have prepared an input file ahead of time. The file lists the six source plates and the wells from each plate.
I’ll take a look at the input file later when I get into the protocol.
I also have to let the software know, through a format file, how I want the source material to be dispensed into the destination plate.
That is, what will the layout be in the destination plate? The software has a wizard that allows me to specify the layout when I create the protocol. So I’ll do that later.
I plan to use my BenchCel Workstation to run the protocol. The workstation looks like this:
I have a BenchCel device at the center that can store and move labware.
I plan to downstack labware from the BenchCel stackers and move them to the Bravo deck for the hit-picking tasks.
I will use the Multidrop device to fill the destination plate with diluent immediately after the hit-picking tasks are finished.
I will then move the destination plate back to the Bravo deck for the serial dilution and plate replication tasks.
Labware will move back to the BenchCel stackers after their processes are finished.
That’s the overall flow of events. However, before I do anything in the VWorks software, I need to have a detailed plan that will specify where labware will be in the BenchCel stackers
and on the Bravo deck. This will facilitate protocol writing and prevent deadlock and other errors.
After careful planning, I came up with this design. The destination and source plates will be stored in BenchCel stackers 1 and 3.
When they are finished, they will return to stackers 2 and 4.
The tipboxes for serial dilution will be stored in stacker 5 and return to stacker 6. Note that I will reuse the same tips for the replication tasks.
On the Bravo deck, I plan to store the tipbox for hit-picking at location 2. An empty box will reside at location 3 for used tips.
Lastly, the plates for replication will be stacked at location 1. Note that deck location 9 is reserved for labware handoff.
That is, the BenchCel robot can place labware or pick up labware at this location.
During hit-picking, the destination plate will downstack from stacker 1, move to location 9, and then move to location 6.
The source plate will downstack from stacker 3, move to location 9, then move to location 5. Samples will be transferred using the tips at location 2.
Used tips will be removed at location 3. As each source plate is finished, it will return to stacker 4. So far, so good. No location conflicts.
After the sample transfer is finished, the destination plate will move to location 9, then go to the Multidrop device. After the diluent is added, the plate will return to deck location
9, then move to location 5. This is possible, because location 5 is no longer occupied by the source plate.
Why I decided to move the destination plate to location 5 instead of returning it to location 6 will be explained in a minute.
Again, so far, no location conflicts.
During serial dilution, the tipbox from stacker 5 will go to location 9, then move to location 7. So there is no location conflict.
Before pressing on the tips for serial dilution, the pipette head will be set to perform column-wise liquid operations.
I plan to use channels in the A1 column. With this selection, only some deck locations are accessible for
the serial dilution tasks, such as location 5. Location 6 will not work, because the pipette head cannot move to columns 2 through 24 in the destination plate.
That’s why I moved the destination plate to location 5.
For more information about accessible Bravo deck locations based on pipette channel selections, see the VWorks Automation Control User Guide.
And finally, during the replication process, the plates for replication will downstack from Bravo deck location 1 and go to deck location 8.
Location 8 is not occupied, so there is no location conflict.
Note that before the second replication can downstack from location 1, the first replication plate must upstack to location 4
so that the second replication plate can be placed at location 8.
Again, everything works out. There is no location conflicts in replication. So the labware setup is good to go.