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Cancer Institute
Microscopy Core Facility
Z
A
I
A1
M1
EISS
XIO
MAGER
AND
A U
G
SERS
UIDE
This is intended to provide a basic coverage of use of the Zeiss Axio Imager M1 for multi-
channelled fluorescent acquisition. For more complex usages please contact
Fig.1 – Front View of Microscope
Viewing
Slide
Microscope
Ocular Viewer
control
touch-screen
Mercury Lamp
(Reflected light)
Main Stage
Control Box
Microscope Power
st
1
Focusing
Supply Box
Wheel
V1.0
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Related Manuals for Zeiss Axio Imager A1
Summary of Contents for Zeiss Axio Imager A1
- Page 1 Microscopy Core Facility EISS MAGER SERS UIDE This is intended to provide a basic coverage of use of the Zeiss Axio Imager M1 for multi- channelled fluorescent acquisition. For more complex usages please contact Fig.1 – Front View of Microscope Viewing Slide...
- Page 2 Cancer Institute Microscopy Core Facility 1) S WITCHING ON . 30 HECK ICROSCOPE OOK OR WITH PREVIOUS USER RIOR TO WITCHING ON 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.
- Page 3 Cancer Institute Microscopy Core Facility Fig 3 – Side View of Lower Microscope Focusing Wheel TL aperature controls Stage Returning Button Stage Dropping Button 2) L OCATE 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.
- Page 4 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.
- Page 5 Select Experiment tab and click on ReUse button – Fig 6 Select a .zvx file normally found in directory; ‘my documents/Carl Zeiss/data/experiment’ These files are labelled with the combination of channels that will be examined by microscope if chosen. For example in the Experiment box shown in Fig 6 the name shown is ‘dapi-gfp-rhod-phase*’...
- Page 6 Cancer Institute Microscopy Core Facility Fig 7 – C tab of Multidimensional Acquisition Ensure the correct condenser filter is selected for application – Fig 8 H – Brightfield 2 – Phase contrast in x20 objective 3 – Phase Contrast in x40 and x60 Objectives I,II and III are for DIC applications which are not currently installed, treat as brightfield...
- Page 7 Cancer Institute Microscopy Core Facility If you do not desire to collect a Z-stack ignore sections j) to o) j) Open the Z tab – Fig 9 Fig 9 – Multidimensional Acquisition Z tab if you desire a collection of images through the Z axis click on Z button at top of tab –...
- Page 8 Cancer Institute Microscopy Core Facility 4) I MAGE ROCESSING NNOTATION IEWING YOUR IMAGE OR TACK You should now have a Z-Stack of you sample in every channel desired - Fig 10. In this format the data is still raw and is best saved to be processed separately afterwards. Fig 10 –...
- Page 9 Cancer Institute Microscopy Core Facility Choose a view with each channel tiled separately using gallery view. ECONVOLUTION OF TACK Fig 11 – Deconvolution Workarea Axiovision software comes with deconvolution image processing option. The function of deconvolution is to ‘clean up’ images to remove positional and chromatic aberrations.
- Page 10 Cancer Institute Microscopy Core Facility RTHOVIEW Fig 12 – Orthoview Workarea The Orthoview option will take a Z-Stack and average and overlay images so that all the information from the Z-Stack can be viewed at once in a 2D image. a) select image to process b) select initial and final frame of view desired and enter into parameters...
- Page 11 Cancer Institute Microscopy Core Facility NNOTATIONS Axiovision can annotate images for publication. Things like Text, Arrows, Scale bars and geometric shapes can be added to your choice of colour, size and style. These can be chosen from the annotations tab in the tool bar – Fig 4 Fig 14 –...
- Page 12 Cancer Institute Microscopy Core Facility Fig 16 – Annotation / Image Properties Window Figure 16 shows the properties window that can be accessed for any image by right clicking on the image and selecting properties. It is in this window that the Attributes tab can be selected where properties of annotations can be changed.