Cancer Institute Microscopy Core Facility
Ocular Viewer
Main Stage
Mercury Lamp
(Reflected light)
Control Box
Microscope Power
Supply Box
Microscope
control
touch-screen
Viewing
Slide
1st Focusing
Wheel
ZEISS AXIO IMAGER A1 AND M1
A USERS GUIDE
This is intended to provide a basic coverage of use of the Zeiss Axio Imager M1 for multichannelled fluorescent acquisition. For more complex usages please contact
Fig.1 – Front View of Microscope
V1.0
Cancer Institute Microscopy Core Facility
Axiocam
CCD image
capturer
Viewing
Slider
Objectives
Main
Stage
XY
adjust
RL Natural
Density filter
wheels
RL aperture
control
Reflected light
(RL) centering
bulb
Slide
Holder
RL Glare
block
Transmitted
Light (TL)
adjustment
wheel
Ocular
Viewer
height adjust
1) SWITCHING ON
CHECK MICROSCOPE LOG BOOK OR WITH PREVIOUS USER PRIOR TO SWITCHING ON. 30 MINUTES MUST
HAVE PASSED BETWEEN TURNING OFF REFLECTED LIGHT LAMP AND TURNING BACK ON IN ORDER FOR
MERCURY BULB TO COOL.
a) Switch on mercury lamp control and Microscope power boxes located
proximally to main microscope stage. Turn on Microscope switch (located
behind 1st focusing wheel)
b) Turn on computer and Activate AxioVision Program
Fig 2 – Side View of Microscope
V1.0
Cancer Institute Microscopy Core Facility
2nd Focusing
Wheel
TL aperature
controls
Stage Returning
Button
Stage Dropping
Button
Fig 3 – Side View of Lower Microscope
2) LOCATE FOCAL PLANE OF SAMPLE
a) Using the microscope touch-screen open TL (Transmitted light) filter
b) Reduce light level using TL adjustment wheel
c) Place sample slide in slide holder.
d) Select magnification
if using x40 or x60 addition of droplet of oil is necessary. This is
achieved by pressing stage dropping button and adding a tear sized
drop on top of slide (so cover slips are necessary) and returning
stage by pressing stage returning button. Make sure at this point to
press ‘set working position’ on the touch screen control.
e) Using Ocular Viewer (viewing slide must be fully inserted) focus the sample
using Focusing Wheel. (For finer focus use focussing 1st focusing wheel)
f) Switch TL filter to closed on touch-screen controls
3) ACQUISITION OF IMAGES
V1.0
Cancer Institute Microscopy Core Facility
It is possible to choose various methods of acquisition of an image and most
actions capable from the software are possible. This will show you how to collect a
Z-Stack. If you only desire a single multichannel image you can leave out to portions
about Z-stack. Remember to not select a Z-stack (section f) )
Fig 4 – Axiovision Tool Bar
a) Select Workarea tool bar if workarea is not showing – Fig 4
Fig 5 – Work area
b) Select Objective of choice (ensuring
oil is used as previously described
above in 2c) or leave as presently
chosen - Fig 5
c) Select ‘Multidimensional Acquisition’
sub tab (as shown also in Fig 6)
V1.0
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