Olympus IX70 Panduan Pengoperasian - Halaman 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

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