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solutions must also contain an antagonist, you can have 1 antagonist concentration and 4 agonist
concentrations with that one antagonist concentration in one protocol file. To test 4 antagonist and 4
agonist concentrations, you would need to link 4 protocol files (Section 10.3.2).
10.3.1.3 Fast0 Perfusion Example - Testing 4 Agonist and 4 Antagonist Concentrations
An example of applying 4 different agonists and 4 different antagonists (one of which could be no
antagonist, ie ACSF) when the antagonist does not rapidly unbind (Table 10.3.1.1.1) is shown in Fig.
10.3.1.3.1. The protocol starts out with the Fast0 Perfuse event in the Protocol Builder causing Ch 1,
Antagonist 1 to be perfused. After a delay, Ch 5, Agonst 1 is applied for 1 second during the P0sweep.
Then shortly thereafter Ch 6, Agonist 2 is applied during a P1sweep, Ch 7, Agonist 3 is applied during a
T0sweep, and Ch 8, Agonist 4 is applied during a T1sweep. Then the perfusion solution is changed to Ch
2, Antagonist 2, and application of the 4 agonist solutions is repeated. Then this is repeated for Ch 3,
Antagonist 3 perfusion, and finally for Ch 4, Antagonist 4 perfusion.
Fig. 10.3.1.3.1. Applying 4 different agonists and 4 different antagonists when the antagonist does not
rapidly unbind. The Fast0 Perfuse events (shown in red in the Protocol Builder on the left) set the
ongoing perfusion to Ch 1 Antagonist 1, Ch 2 Antgonist 2, Ch 3 Antagonist 3 and Ch 4 Antagonist 4. The
agonist solution is applied during the sweep. During the first 400 msec epoch, Step0, and the third epoch,
Step2, a -1 in the Amplitude (V) field (see red rectangles) means that the current perfusion solution set by
the Fast0 Perfuse event in the Protocol Builder is maintained in that part of the sweep. During the 1000
msec epoch, Step1, a voltage from 1 to 8 in the Amplitude (V) field (black rectangle) sets the Ch from 1 to
8, in this case for T1sweep 8V=Ch8 or Agonist 4. AO1 Gain is 1. To see this connect AnalogOut1 (AO1)
to AnalogIn0 (AI0 or AD0).