Olympus IX70 Руководство по эксплуатации - Страница 29
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Current
acquired
image or
reloaded data
required always). Save the calibration for future use (recall using LOAD button). You can exit the dialog
("OK"), and see a titration curve by clicking on "Titration" (even if it's already checked). It's good to
make sure the system can give back the reference value for a standard following calibration.
It's ideal to correct images for uneven illumination and backgrounds. This can be done using the
REFERENCES sidemenu button. A blank chamber or area from your sample can be grabbed for
background using the "Acquire" button (IMAGE mode). You can also use a ROI as background, if drawn,
to compare values across images vs across time at the same location.
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Pick
load/save
locale
changing the pulldown list. This is useful if drugs
might alter background during an experiment.
Flat-field or SHADING correction eliminates
differences in illumination quality by dividing
images by a correction standard. You need a
uniformly fluorescent standard, like a drop of
standard, and set the camera to near saturation
(but nothing saturated) for each wavelength. You
might get by with one reference if the illumination
pattern is similar for two wavelengths close
together. Be sure to check the 'do it' boxes to
actually turn on corrections.
the image data
you save/use, so do set this up
correctly. Ratios tend to decrease illumination
artifacts, you really should do these for single
wavelength dyes or experiments where you want
29
Use the
Acquire
Standards Dialog
to create or load
calibrations into
the system. Place
the standard on
the scope, enter
the value on a
line
(note double
>> indicating
ACTIVE ENTRY
LINE, change by
clicking on line to
. Enter the
use)
values in proper
order low to
high. Hitting
"Acquire" stores
the buffered
standard image.
Repeat for all
standards (at
least two are
Corrections will alter