Olympus IX70 Instrukcja obsługi - Strona 6

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can also be layered on top of the media to prevent evaporation without image distortion. DO NOT USE

SILICON OIL WITH THE WARNER DISHES! It will ruin the rubber gaskets and cause leaks.

We have added a transmitted light shutter to gain better control of DIC capture and

fluorescence sequences. The brightfield bulb can be turned on with the power switch and button and set

to the desired brightness with the intensity slider, and then controlled with the shutter. The shutter opens

automatically when the DIC button is selected and closes when a fluorescence button is selected or once

an image is collected. There appears some resistance of program control vs. manual microscope control

where you may have to hit the power-on button and scroll voltage up and then try the power button again

to get the lamp to behave (it will perform once you show it who's boss, I guess; It's a MM bug. This isn't

an issue in MetaFluor software).

Fluorescence:

It's important that no other electronics be powered up when firing the arc lamp; it can

create a power surge that will fry other components (a very bad thing). Since this is a manual microscope,

choices in the software require changes to the microscope that must be made by hand. We have added

pop-up window prompts to MetaMorph to guide you in changing filter cubes, etc. Fluorescence uses

excitation filters to excite your probe(s) with a narrow band of wavelengths, a dichroic mirror to reflect

the excitation onto your sample while allowing the longer fluorescence to pass trough, and a barrier (AKA

emission) filter to block stray excitation light and limit the output color. Most filter cubes use dedicated

excitation and emission (barrier) filters. This is the case with the DAPI, FITC (wide) and Cy3 cubes in

the turret. These filters will work for both the camera and the oculars (by eye or by image). A few cubes

(like fura-2, and the cubes in the "FRET" set) require proper exciters or barrier filters to be set by the

filter wheels (Sutter controller) through the software. These filters also work by both by eye or by camera.

However, the new quad mirror requires the filter to be changed thought the software. It will image DAPI,

FITC (narrow), Cy3 or Cy5, depending on the taskbar button selected, but it will ONLY work through the

camera. You will NOT see anything by eye but a lot of very bright light!! Use the single color cubes to

find your sample and only use the quad if you need Cy5 or if you are capturing multiple wavelengths with

the Acquire Multiple Wavelengths dialog. Regardless of the filters used, it's highly recommended

that changes of all filter cubes (for the color of fluorescence) be initiated through the taskbar.

To choose a color channel, click the taskbar button for the wavelength you want, then

follow the on-screen prompt or filter list to rotate the manual filter turret under the objectives to the

correct position (see "filter wheel," Fig. 1 and 2). Press "Continue" on the pop-up window to toggle the

shutter (if necessary). Alternatively, you can learn the settings of the Sutter Lambda 10-2 filter changer

box located to the right of the microscope (long, bottom-most controller with keypad). Be aware that not

setting the exciter or emitter filters correctly can blast your sample, or worse, your poor eyes, with the full

arc lamp output. The Sutter filter changer operates a shutter to

limit illumination to actual image collection (to minimize

sample bleaching and phototoxicity), a high-speed excitation

filter changer, and a high-speed emission (camera-side) filter

changer. {optional reading /details}

control (parallel port). Manual mode requires you to press the "Local"

button on the top right of the keypad. Press either "F1" of "F2" to select a

filter changer to command. Note that the shutter is on F1; pressing the

"shutter" will toggle the light on/off. The other keys address the ten filter

positions per wheel (numbered 0 through 9).

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Fig. 4 Neutral density filters