Molecular Devices ImageXpress Confocal HT.ai, Руководство пользователя

Страница: 56 / 256

ImageXpress Confocal HT.ai High-Content Imaging System User Guide
56 5084773 A
Fluorescence Background
There is a large difference in auto-fluorescence between glass and plastic. There can be up to
a five times difference in auto-fluorescence among plates from different manufacturers.
Bottom Thickness
Compare the thickness of the plate bottom with the working distance of the objective lens to
ensure that it is compatible. In general, objectives with higher numerical aperture (NA) tend to
require thin-bottomed plates. Objectives with application-optimized correction collars are
compatible with a larger range of plate thicknesses but tend to have lower NA. Plates with a
bottom thickness comparable to a standard coverslip (0.17 mm) work well with all supported
objectives.
Note: Plates with ultra-thin bottoms or very thick bottoms can be more uneven and can
cause focusing issues. For best results, use an imaging-quality plate with a bottom
thickness between 0.15 mm and 0.7 mm.
Plate Flatness or Reproducibility of the Z-Pattern
A flat plate is faster to scan than an uneven plate. The reproducibility of a plate allows you to
set tighter focus ranges specifically for that plate type. This reduces the amount of focusing
needed and speeds up acquisition. The major component in plate flatness is the variation from
a well to a neighboring well.
Plate Skirt
With short working distance objectives, imaging the edge and corner wells can cause the
objective to bump into the plate skirt, which requires you to omit the outer wells from the
experiment. Some manufacturers offer low-skirt or no-skirt options that allow you to use all the
wells in the plate with a short working distance air objective.
Batch-to-Batch Consistency
Some plate manufacturers are more consistent in producing plates than others. If parameters
such as the plate-bottom thickness vary from batch to batch, you must optimize the plate
settings for each batch.
Robot Compatibility
If you use a plate-loading robot, ensure that the plates can easily be held by the robot
grippers. Some types of plates do not work well with the grippers supplied with the robot and
require custom grippers to work correctly. If one or more plate types do not work with your
robot grippers, contact Molecular Devices for assistance. Also make sure that there is a
consistent location to affix a barcode on the side of the plate.