Directions for Zeiss Axiovert 200M
START UP
1.
Turn on mercury lamp controller (#1). Wait a few seconds to make certain lamp is
on.
2.
Turn on the x-y stage controller (#2)
3.
Turn on the camera controller (#3)
4.
Turn on the microscope (#4) (green button on the right side of the microscope.
5.
Login to the computer with your CNET ID and password. Double click on the
Openlab program on the computer.
IMAGING
1.
Choose the correct stage for slide or dish. Make sure the stage is properly inserted
(there is a spring in the corner). The stage should be laying level: Don’t have one
side higher than the other- it should not wobble. If your sample is on a slide, make
sure the coverslip is facing down. Choose the objective you want by moving it in
to place or choose the objective on the Microscope palette. The dry objectives are
10x, 20x, and the first 40x. If you are using an oil objective (the second 40x, 63x
or 100x) put 1 drop of oil directly onto the objective. When focusing with the oil
objective you should have a space of oil between the objective and the cover slip.
The objective and cover slip should NOT be touching.
slide
Coverslip
2.
Check the fluorescent palette in the program: if the filter you need is not in place,
use the red- handled Zeiss tool and unscrew the screw that holds the reflector
turret in place with 2 turns. (DON’T unscrew until the screw falls out). Gently
push the reflector turret out and exchange filter cubes. (Do not touch the actual
filters with your fingers). The cubes should all be level to each other if properly
inserted in the turret. Carefully replace the reflector turret (it should be smooth
and easy to insert- do not force it! If you are forcing it, then you are inserting it
incorrectly, and it will get jammed!) Use the Zeiss tool to lock in the reflector
turret after changing your filter. Then change the label for the filter in the reflector
pallet in the program.
3.
If possible start by imaging your sample using
transmitted light
to prevent
bleaching. Click on Transmitted Light or the dic (#5 position) in the Zeiss
reflector palette and focus using the focusing dial on the right side of the
microscope.
(Do not open the fluorescent shutter at any time while on
transmitted light!)
Use Koehler illumination to focus the condenser for
transmitted light every time you switch to a new objective. For transmitted light,
choose the camera shutter that sends 50% of the light to the camera and 50% of
the light to the ocular.
a.
Koehler Illumination: After focusing on your sample with transmitted
light, close the diaphragm, and use the condenser-focusing knob on the
right side of the transmitted lamp to focus the condenser unit so that the
diaphragm is sharply focused. You should now see your sample and the
Oil objective
oil
