HP D300 Operating Manual page 46

Figure 5-19 Plate Layout Wizard Titration1 pane
Click here
to move
between
panes.
Or use the
Back and
Next
buttons
here.
Define the titration to be used for the selected fluids:
c.
i. Type a number in the box to specify how many replicates of each concentration or
volume will be used for each fluid as you did in the
wells‖ section of this document. For example, if triplicates are desired for each fluid,
enter 3.
ii. Click the arrow in the drop-down list box and select a value to define how many titration
zones the titration will have.
Titration zones are regions (concentration ranges) of the titration which have the same
type of spacing between concentrations.
If a titration is desired to have tight spacing around the expected IC50 value, with more
sparsely spaced concentrations on either side of the IC50, a three-zone titration could
be chosen. However let's practice with a simple one zone titration for eight fluids, four
from the existing fluids table, and four new fluids. The eight fluids have already been
selected with the Use check boxes.
iii. A twelve-point singlet will be defined for each of the eight fluids. Click in the Low (µM)
and High (µM) columns of the table to enter the concentration endpoints of the titration,
then click in the Titration Type column to define how the points are spaced between the
low and the high concentration limits, and then type in the number of points. If the
titration type is not Linear or Log, then the spacing between concentration points is
fixed, so the number of points, the high and low limits are adjusted by the software to
make a valid titration series when a fixed spacing is specified.
iv. The Valid Concentration Range specifies the concentration limits for these. The lowest
possible well concentration is determined by the minimum drop volume, the source fluid
concentration, and the assay volume. The maximum well concentration is determined
by the DMSO limit and the fluid concentration.
v. At the bottom of the dialog box, the endpoints and the point spacing are shown
graphically. Click the box labeled Log to toggle between log and linear views of the
layout.
Click Next three times or click Positioning: Titrations to open the Positioning: Titrations
d.
pane. Only a single titration1 and no titration2's will be used for this example. The twelve
point singlet for each of the eight fluids selected fills up the entire 96 wells.
Type a number here to specify
the number of replicates
HP D300 Digital Dispenser
Select a value to define
the number of zones in
the titration
Define the titration here
―Set concentrations or volumes in
Valid
Concentration
range
Click here to
toggle between
log and linear
views
46