TITLE: Dispensing agar into multiwell plates July 2019 updates
AUTHORS: Ida Barlow

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Protocol for pouring agar into 96 well plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60</span><span style = "vertical-align:super;">o</span><span>C 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. </span></div></div>

[STEPS]
?. [Configure Integra VIAFILL]
Insert large cassette (suitable for dispensing volumes from 5-9999μL) into the machine:
?. [Configure Integra VIAFILL]
Configure X, Y, and Z settings for the multiwell plate by clicking on tool symbol -> stage alignment -> 96 well 8Ch.
For UNIPLATE96SQWLF 650U:X = 95.6Y = 4.2Z = -22.5
?. [Configure Integra VIAFILL]
Exit settings by pressing the back button
?. [Dispensing Agar]
Press on the program you wish to use (see later for configuring your own program)
?. [Dispensing Agar]
Make sure that the correct cassette (8 channel large) is listed and change if necessary
?. [Dispensing Agar]
Select the volume you wish to dispense
For 96WP:150 μL
?. [Configure Integra VIAFILL]
Prepare a 250ml bottle of hot milliQ water in the microwave and keep in the 60oC waterbath along with the agar. The water is important to have on hand in case of tubing blockages.
?. [Configure Integra VIAFILL]
Put the plate into the stage and then press 'Move' so that the plate moves so that it is under the dispensing cassette. The pipette tip closest to you should be over well 7H
?. [Configure Integra VIAFILL]
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.
?. [Configure Integra VIAFILL]
Use the X, Y arrows to move the plate so that the pipette tips are centered in the middle of column 5.
?. [Configure Integra VIAFILL]
Save all settings.
?. [Dispensing Agar]
Place the end of the tubing from the casettee into the agar that is being kept warm in the water bath set to 60oC
?. [Dispensing Agar]
Press 'Prime' to prime the tubing and allow to finish so that agar flows from the pipette tips.
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.
?. [Dispensing Agar]
Select 'set height' and set the appropriate height for tip height when dispensing (usually all the same as the Z height determined earlier)
?. [Dispensing Agar]
Place a clean plate in the stage
?. [Dispensing Agar]
Press run and then plate should fill with agar
?. [Dispensing Agar]
Repeat steps 11-12 until all the plates have been filled.
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]
'Recover' to remove all the agar from inside the tubing.
?. [Cleaning the cassette]
Place the tubing ends into the hot water.
?. [Cleaning the cassette]
Prime so that the water runs through and clears all the agar
?. [Cleaning the cassette]
Reverse prime to remove the water
?. [Cleaning the cassette]
Release tension from the tubing and remove cassette
?. [Cleaning the cassette]
Double wrap the cassette in aluminium foil/place in tip box for autoclaving
?. [Configure Integra VIAFILL]
The pipette tip end slots into the dispenser holder over plates
?. [Configure Integra VIAFILL]
The first plastic tube holder is placed on the metal plate on right hand side of the peristaltic arm
?. [Configure Integra VIAFILL]
The second plastic tube holder is fitted onto the metal plate on the other side of hte peristaltic arm. Make sure the metal pins on either side slot into the holder
?. [Configure Integra VIAFILL]
Flip the metal plate arm up so that it is in place
?. [Configure Integra VIAFILL]
Secure everything in place by flipping up the cover