Feb 26, 2024
Biomek 96 well plating
- William Buchser1,
- jwaligor 1
- 1Washington University in St. Louis
- WashU FIVE @ MGI
Protocol Citation: William Buchser, jwaligor 2024. Biomek 96 well plating. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlko5m1v5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 21, 2023
Last Modified: February 26, 2024
Protocol Integer ID: 83801
Keywords: randomized plating of cell, plating randomized plating, using biomek, cell
Abstract
Randomized plating of cells using Biomek.
Creating Plate Layouts in FIVTools
Open FIVTools and click "Layouts."
Edit "Exp Name" to the correct FIV Number (make sure to keep "FIV" in the name)
1. Click once on the “Export Folder” bar so it updates the location
Choose the desired number of 96-well plates by typing 1, 2, 3, or 4 in "Count."
- Select “96 Mote” to exclude plating from the outermost wells
- Select “384 Full” for a 384 well plate
- If the layout is not standard, adjust the “First Row”, “First Column”, “Last Row”, and “Last Column” boxes appropriately
- If doing an antibody plate, check the “Ab Plate” box and:
- Make “Count” = 0 for one antibody plate
- Make “Count” = 2 for two 96-well plates and an antibody plate
Change “Cells per Well” to desired plating density.
Copy experimental condition names (often genotypes, dosages, G1/G2, etc.) and click “PASTE Names” to auto-fill the 12-well source plate.
- “Name” can also be manually filled
- The “Well” column can only have row/column combinations of a 12-well plate (A-C, 1-4)
- “Relative Representation” allows you to assign different weights for representation in the 96-well plates
- Use the underscore symbol to have a cell type only show up in the antibody plate layout (for example: “_cell type”)
Click "Fill Plates"
1. Try to make sure the empty wells are not on any edges and if doing multiple plates that they cannot be flipped or rotated to the same location as another plate.
Go into the “4 Mapping” folder of the respective experiment and make sure there is a PlateMap, PlatingSetup, and script.
Setting up Biomek
Find the correct method.
The algorithm will look like this:
Set up the deck in the configuration shown
Make sure the two tip rows closest to the front are full in the “tip_box” and the “empty_1” and “empty_2” are completely empty.
Only use the number of 96-well plates needed for your experiment and place them in order starting with “0.”
- Keep the lids on the plate until you are ready to begin the Biomek program.
Open the appropriate experiment folder, go to “4 Mapping” and open the “FIVXXX_script” file.
Copy the entire script.
Go to the “Scripted Let” step in the algorithm and paste your entire script.
- Scroll up to the top to make sure the correct experiment number and an appropriate time stamp are there
Go back to the “4 Mapping” folder and drag and drop the “FIVXXX_PlatingSetup” file onto and EXCEL sheet (DO NOT OPEN AS TEXT FILE).
After creating cell suspensions for each source condition and obtaining counts, input their counts into the appropriate positions of the “Cells/uL” column:
With a new 12-well plate, set up the Source Plate by adding the volumes in the “uL Media” and “uL Cells” columns into the appropriate wells:
Place the Source Well in the correct position on the deck,open the lids to all plates, and close the front door.
Press “Run” (the green arrow) and confirm the configuration in the pop-up box.