Molecular Devices ImageXpress Confocal HT.ai, Руководство пользователя

Страница: 104 / 256

ImageXpress Confocal HT.ai High-Content Imaging System User Guide
104 5084773 A
Option Description Notes
Auto
Correction for
TL This is a shading algorithm tuned for
transmitted light but can be used for
fluorescence. It performs shading
correction but not background
subtraction. This algorithm is faster
than the Auto Correction for TL
Legacy algorithm, and does not rely
on the TL Legacy Shading Correction
Refinement Level on the Wavelengths
tab. The shading correction is
calculated individually for each image.
No reference image is used. Not recommended for montages.
Not recommended for Z-stack
assays.
Not recommended for time lapse
assays.
Auto
Correction for
TL Legacy Images are corrected for shading
automatically, without a need for
reference images. The shading
correction is tuned for transmitted
light, and it adjusts for uneven
illumination of the sample. The default
value for the TL Shading Correction
Refinement Level (Wavelengths tab) is
2. The higher the refinement level,
the flatter the transmitted light will be,
but the longer the correction will take.
The shading correction is calculated
individually for each image. No
reference image is used.
Not recommended for fluorescent
images.
Not recommended for montages.
Not recommended for Z-stack
assays.
Not recommended for time lapse
assays.
Legacy
Correction Images are corrected for shading
using reference images specific to
objective/filter combinations (and
possibly other optics, such as
confocal disk). The reference images
must be created in advance by the
user. They are typically stored in
C:\Shading Images but may be stored
elsewhere on the computer.
Can be used for either fluorescence or
transmitted light. See
Using Legacy
Correction Image Shading on page
105
.
In general, we recommend the
automatic methods over the Legacy
Correction method because
correction is based on the current
state of the system and is not subject
to variation over time.
Recommended for montages.
Not recommended for multi-site
imaging of plates in transmitted
light, particularly at low
magnification, since the meniscus
causes site-specific illumination.
Shading reference images must be
updated when there are changes to
the optics, such as the light source
or the liquid light guide.
May be inconvenient to create
reference images for oil-immersion
objectives.
Table 5-10: W# Tab: Shading Correction Options (continued)