23
Figure 28: Protocol page
This means that a step with
Rep#
= 5 and
Time
= 20secs will dispense the selected solutions for 20 seconds,
then move the collection tray to the next row of wells and dispense the same solutions for another 20 seconds,
and so on for 5 consecutive well rows. So in general, the total dispensing time in a step would be the product of
Rep#
and
Time,
and distributed evenly in consecutive well rows equal to
Rep#
.
The next column in the protocol is the
Flow-rate (µL/min)
in micro-liters per minute. This determines the rate at
which the solutions are supplied to the cells during the step. This parameter controls the speed of the peristaltic
pump. All channels will have the same flow.
The next four columns (A, B, C, D) relate to the automatic fluid handling system (see section 5.2). In these fields
you can assign the solution (1 to 8) for each of the channels. If you are not using one of the channels, assign a
zero "0" to the solution field.
The next and last column is the
Pause
option. This parameter will determine if the machine pauses after a step is
finished. This is an advanced option, where you need to perform a manual operation between the steps. For all
conventional Perifusion experiments, this option should be set to
NO
.
On the top right of the screen you will find a
# of Plates
. This number will tell you the number of well plates
required to complete the protocol. This will help you have all the plates in hand to avoid unnecessary delays.
Creating a Protocol
To create a protocol, click on the "NEW" button. Click "ADD STEP" button to add a step to your protocol. A row
filled with "X" will appear. Replace all the X with your protocol values. No X can be left on the protocol. Repeat the
process to add more steps. Assign a name to the protocol and click "SAVE" to store your protocol in the
machine's memory for future use. You will now see the name of the new protocol displayed on the left pane.
Loading a Protocol
Biorep can provide you with a spreadsheet template that will allow you to create protocols from the comfort of
your office. Simply fill out the template as you would on the Perifusion touch-screen and save the protocol onto a
USB stick as a tab delimited text file. On the perifusion system, plug the USB stick to one of the ports in the back
of the machine; Click the "LOAD" button on the wizard and navigate to the txt file you created. The protocol will be
loaded into the wizard interface and will be ready to save and execute.
7.3
System setup
Before an experiment can be performed, the machine has to be properly set-up and prepared. The basic steps to
complete are the following:
Chamber setup
Tubing setup
Source priming
Perifusion chamber cell loading
Pre-heating the incubator/ Cooling the sample tray
Summary of Contents for PERI-4.2
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