Jan 17, 2024
Dispensing agar into multiwell plates
Forked from Dispensing agar into multiwell plates
- 1MRC LMS, Imperial College London
Protocol Citation: Bonnie Evans 2024. Dispensing agar into multiwell plates. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwd759lmk/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: January 11, 2024
Last Modified: January 17, 2024
Protocol Integer ID: 93386
Abstract
Updated protocol for pouring agar into multiwell plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60oC waterbath until ready to dispense, and whilst dispensing. The X, Y, Z positions for dispensing can be adjusted according to the multiwell plates being used.
Materials
MATERIALS
Whatman UNIPLATE 96-Well Clear MicroplatesVWR international LtdCatalog #WHAT7701-1651
Configure Integra VIAFILL
Configure Integra VIAFILL
Prepare a 250ml bottle of hot milliQ water in the microwave and keep in the waterbath along with the agar. The water is important to have on hand in case of tubing blockages.
To pour 6, 12, 24 or 48 well plates, disconnect the appropriate tubing from the cassette. Tube 1 is closest to the instrument. Select 96 well plate in programme.
Insert large cassette into the machine
Configure X, Y, and Z settings for the multiwell plate by clicking on tool symbol -> stage alignment.
Note
For UNIPLATE96SQWLF 650U:
X = 95.6
Y = 4.2
Z = -22.5
Put the plate into the stage and then press 'Move' so that the plate moves so that it is under the dispensing cassette.
Use the up and down arrows to move the pipette tip so that they hover just over the plate and make note of the height (this will be entered into the dispensing program at a later step). Press 'Fast/Slow' button to switch between fast and slow movements.
Use the X, Y arrows to move the plate so that the pipette tips are centered in the middle of column 5.
Save all settings.
Exit settings by pressing the back button
Dispensing Agar
Dispensing Agar
Press on the program you wish to use (see later for configuring your own program)
Make sure that the correct cassette is listed and change if necessary
Select the volume you wish to dispense
Note
For 96WP:
200 μL
For 24WP:
600 μL
For 6WP:
4000 μL
Select 'set height' and set the appropriate height for tip height (usually all the same)
Place the end of the tubing from the cassette into the agar that is being kept warm in the water bath
Press 'Prime' to prime the tubing and allow to finish so that agar flows from the pipette tips.
Note
IMPORTANT:
Once the agar is in the tubing it is important to act quickly to avoid agar solidifying and causing blockages. If you are particularly concerned about agar cooling in the tubing, wrap the tubing in aluminium foil to keep hot.
Unblocking the tubing9 steps
If the tubing does block, clear the blockage by 'reverse priming' as much of the agar as possible.
Then place tube ends in the hot water and prime continuously with hot water until the water runs all the way through.
If you are having trouble getting the water through, squeeze and massage parts of the tubing where you can see blockages to force the agar along and allow the water to pass.
Once all cleared, 'reverse prime', and reprime with the agar
Place a clean plate in the stage
Press run and then plate should fill with agar
Repeat steps 11-12 until all the plates have been filled.
Note
Little drops of agar can solidify on the tip ends. It is often good to remove these drops using a pipette tip every few runs so that blockages do not occur.
Cleaning the cassette
Cleaning the cassette
'Reverse prime' all the agar
Place the tubing ends into the hot water.
Prime so that the water runs through and clears all the agar
Reverse prime to remove the water
Release tension from the tubing and remove cassette
Double wrap the cassette in aluminium foil for autoclaving