Olympus IX70 Bedieningshandleiding - Pagina 5

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decoded by the sheet taped to the side of the scope. If you need filters not currently installed, please ask

for assistance in swapping out the cubes (we have more choices than available turret positions). A manual

shutter slide is also located on the side (Fig. 2 "manual shutter"). Push toward the wall to block

fluorescence (but be aware this will prevent you from snapping a picture too!). There is a 1.5x tube lens

that will enlarge the image destined for your eyes (see "mag changer" Fig 2). However, the image quality

is poor and a bright reflection renders images in the camera port in need of post hoc corrections. Use the

1-X (pushed in) position for all quality work. A brightfield lamp is available (above the scope) and is

controlled via the square button on the front left-hand side of the scope base plus the voltage up/down

slider (see Fig 1).

DIC:

The microscope is equipped with Differential Inference Contrast (DIC) optics to generate contrast

in unstained materials. To setup DIC or use is in ocular mode, one needs to use the analyzer slider (must

be inserted under the filter turret) and the optical path should be optimized (Kohler illumination, described

below). The DIC analyzer (polarizer) for image capture is mounted in the emission changer. The visual

DIC analyzer is NOT normally present (i.e., by request) since users tend to leave it in the optical path and

cut fluorescence intensities by mistake (must be pulled out for all work apart from visual work). DIC uses

a pair of prisms, one above the slide, one below. The upper one is found in the wheel on the condenser

Brightfield lamp housing

Field

aperture

Centering

screws

Condenser

aperture

Fig. 3 The condenser

image contrast (at the price of brightness; you will also see more dirt on your slide the more you close it

down).

Note that plastic degrades the DIC effect, and while you may get enough contrast for an image, the

quality is far worse than for cells on a glass coverslip. It is also best to remove the lids from plastic

dishes, especially if there is condensation. A plastic lid with built-in glass window is available. Silicon oil

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(Fig. 3), and should be changed manually to best match the

objective in use. Due to space+cost limitations we have only 3

(upper) prisms: 10x (use for 10x), 40x oil (use for 20x, 40x and

100x), and 60x TIRF/oil (use with 60x lenses). The prisms

match the objectives they are named for, so DIC quality will be

best with 10, 40, 60x lenses, and acceptable with others. The

lower shift prism is constant for all objectives. It has a knurled

screw that allows you change the contrast and apparent

direction of light hitting your sample (also called the shear).

Adjust the screw to get a uniform background and details (see

Fig. 1 "DIC shear adjustment screw"). Before you display the

Focus knob

DIC image, you should optimize the optical path to get the

most even illumination possible. To achieve Kohler

illumination, first find your sample on the slide and make sure

Prism

it is in focus. Then focus the condenser onto that plane by

selection

closing the field aperture (Fig. 3) and using the condenser

focus knob to get the sharpest image of the closed iris leaves

(should be a bright hexagon of light). Use the centering screws

(Fig. 3) to center the iris image, then open the field aperture

iris until it just vanishes from view. The iris on the condenser

itself (Fig. 3 "condenser aperture) can be closed to enhance the