Observing Fluorescence Through The Eyepiece; Acquiring Images With The Improvision Volocity Software; General Informations; Acquiring The Image - Zeiss AXIOVERT 200 Quick Start Manual

Observing fluorescence through the eyepiece

There are 2 options to observe fluorescence: the common way would be to use the filter cubes
in the cube revolver; for some applications the use of a dichroic mirror in the filter cube position
together with the filters in the filter wheels is necessary. The latter setup will only work with the
camera, not with the eyepiece.
•
make sure the excitation / emission filter wheels are in the open position,
unless needed:
•
on the filter wheel pad, select FW1 and then HOME
•
repeat for FW2
•
with the REFLECTOR buttons choose the illumination filter (microscope,
right / bottom behind focus wheel)
•
switch between brightfield and fluorescence
with FL ON / OFF (left / front / top button on
microscope)
•
turn fluorescence immediately off if not
needed, to avoid bleaching!
•
find your sample
Acquiring images with the
Improvision Volocity software

General informations

All microscope parts can be controlled by the software (shutters, filters, focus, ...), except the light
path (BINOCULAR/ FRONT PORT / BASE PORT) and the objectives

Acquiring the image

•
start the VOLOCITY software (make sure the dongle is attached to a USB port)
•
click on the camera icon to activate the live view and microscope controls(for more information
about the Video Toolbar in the upcoming window see Additional Information section)
•
on the microscope stand, change the light path to BASEPORT
•
refocus/ re-centre (if needed, see "refoccussing/ re-centering")
•
adjust the channel presets of the channels you need (see "adjusting channel presets")
•
adjust other acquisition parameters (see "multidimensional acquisition setup")
•
with a click on the camera icon on the video toolbar you capture a single frame with the
settings actually displayed in the video toolbar
•
with a click on the red button icon on the video toolbar you start your recording as defined in
"Acquisition Setup"
Useful: by clicking the "Freeze" button in the software you can keep your actual live image
freezed on the screen; the acquisition stops and the shutters will close, preventing your
sample from bleaching.

refocussing/ re-centring:

o choose the brightfield preset (preset button 1)
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