Olympus IX70 Bedieningshandleiding - Pagina 28

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You can set line styles and axis scales. Defaults

use the ROI color to identify data per ROI and solid vs

dotted to ID the pairs used per ratio. For Fura-2, you

expect the F340 to rise and the F380 to drop if calcium

goes up; if both change the same direction, it's

something else like focus drift, bleaching, etc. You get

ratio plots straight away. You need to calibrate values to

get the real-time calibrated value graph to work.

Calibrate the system, if desired, using defined

buffered solutions and one of several methods offered.

You can use two-points (fully unbound indicator and fully saturated to generate minimum and maximum

ratios, Rmin, Rmax, respectively) and the equation method, or create a titration curve using many

buffered solutions. I prefer an in vitro, multi-point approach for Fura-2, though there are

advantages/drawbacks to all methods their discussion is beyond the scope of this simple manual. Ratio

data are always right but calibrations can yield junk if you're not careful. There is value in saving raw

image data in that you can later apply various calibrations and other corrections (like background

subtractions and flat-field corrections). Enter the dialogs via the top menu CALIBRATION.

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Open the event mark menu by pressing the

Events sidemenu button. You can enter any text you

want associated with an event in the entry box, then

either log it immediately or store it to the reuse list

with buttons at the bottom. Hit the F5 button to

speed mark an event (vs futzing with the mouse);

F6 advances through the list. The list is stored with

your protocol if you save it on closing an

experiment (it will ask you each close).

Graphs can be configured by pressing the

Graphs sidemenu button.