Plate Layout Wizard Advanced Functions; Create Multizone Titration1 Layouts - HP D300 Operating Manual

5-10 Plate Layout Wizard advanced functions

In addition to simple full plate layouts using a single titration for all fluids as described above in
Wizard basics‖, the Plate Layout Wizard can be used to create complex plate designs that would be difficult
to create using the standard graphical user interface described above.
Three types of advanced configurations are described in the following sections. The first of these is where the
titration for a particular compound is not uniformly spaced in log or linear concentrations, which we call a
multizone titration 1 layout. Recall that titration 1 refers to a titration with one compound per well. For
example, the ends of a particular dose response function may show little change in response over a large
range of concentrations, so that these areas do not need to be sampled with as many different doses, while
the region near the IC50 or EC50 point where the response is rapidly changing with concentration could be
sampled with a dense series of closely spaced doses. This would of course require some knowledge of the
IC50 or EC50 prior to setting up the configuration. The process for creating these configurations is described
―Create multizone titration1
in
The next type of complex layout is where in some cases the dynamic range of the dispenser (the range
between the dose from one 13pL droplet to the DMSO limit) is too small for your region of interest. By
specifying your range of interest, the Wizard will help you to determine the fluid pre-dilution required to
achieve the desired dynamic range and set up the dispenser to use two stock concentrations to achieve
greater dynamic range. This type of configuration is described in
layouts‖.
The final type of advanced designs are drug-drug interaction or synergy experiments where two compounds
are placed in the same well for many wells in a set of orthogonal concentrations. We call this a titration 2
layout since there are two compounds in each well, and is described in
per well‖. All of these advanced functions take advantage of the digital nature of your HP D300 Digital
Dispenser which allows ―any dose in any well‖.

5-10-1 Create multizone titration1 layouts

A multizone titration 1 is a set of concentrations or volumes that is not evenly spaced for the entire
fluid. For example, you may wish to create a titration which has more densely spaced points close
to a known or targeted IC50 value, with less densely spaced points further away from IC50 where
there is little change in the response value.
1. To generate a multi-zone titration1, follow steps 1 and 2 in
in the left pane of the Plate Layout Wizard: Titrations and Positioning dialog box. This opens
the Titration1 pane.
2. Select the fluid(s) you would like to use for this multizone titration by clicking in the appropriate
Use box for each desired fluid. You likely will only program a single fluid at a time using this
method since you will center the region of densely spaced doses differently for each
compound.
3. Define a three-zone titration with a tightly spaced zone close to the presumed IC50 of the
compound, and two sparsely spaced zones on either side of the IC50 (see
Click the arrow on the How many titration zones are there? drop-down list box and select
a.
3.
Assume that four wells will be used for the sparsely spaced zones, and eight wells will be
used for the fine spaced zone close to the IC50 value for fluid 1.
In the No. of Points column for zone 1, enter 8, and for zone 2 and zone 3, enter 5. Below,
b.
you will see how this titration which now looks like an 18 point singlet will become a 16
point singlet.
layouts‖.
HP D300 Digital Dispenser
―Create modified range titration1
―Create titration2 layouts: two fluids
5-9-1 above, then click Titration1
―Plate Layout
Figure 5-22).
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