Related Manuals for Zeiss AxioImager A1
Summary of Contents for Zeiss AxioImager A1
- Page 1 The Stereology Workstation Instructions for Zeiss AxioImager A1 Microscope for Stereology By Jon Ekman Basic operation instructions as of 10/1/2009...
- Page 2 Start-up Start-up 1. Turn MASTER switch on Belkin Surge- 1 ..Turn ON Master Switch Master. This will turn on the LUDL stage 2 ..Log on to Computer controller, microscope stand and the 3 .
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Page 3: Koehler Illumination
Align Optical Components Koehler Illumination 6. Adjust contrast using condenser aperture diaphragm slider (4). In most Benefits of Aligning Optics: PROCEDURE: Transmitted Light cases adjust aperture diaphragm slider • Evenly illuminated image. 1. Focus on specimen with the objective to mark matching the NA of the objec- to be used for data collection. - Page 4 Calibration The system is calibrated by ITG staff, and new users get these initial calibration settings at first login and startup. The calibration is done using transmit- ted brightfield method. Calibrate only the objectives you plan to use in your experiment. Recalibrate only if you notice drastic errors in software contour placement between objectives.
- Page 5 Stereology with the Optical Fractionator Workflow 2. Section’s cut thickness: How thick the tissue was cut at. Information below is derived from MBF Bioscience correspondence 3. Section Evaluation Interval: This is the interval between the sections to Tissue Preparation to approach a pilot study is to err on count.
- Page 6 Optical Fractionator Workflow Continued... an area where potential knife damage can occur either by being cut in half, or plucked from the tissue during micro- tomy. In practice, we recommend that 100x-oil). Objective choice will depend top, nucleolus, widest cell perimeter, or the guard zone be a minimum of half the on tissue type and staining.
- Page 7 and past the bottom of the tissue until (hold down the Ctrl key to select more recommend that our customers mea- it is completely out of focus and then than one) and click on View Results. If sure at every site and use this estimate slowly focus back up until something the sections have mirrored contours for their data analysis.
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Page 8: Acquire Images
Acquire Images With StereoInvestigator 1. Koehler align optic train. 2. Deflect light to camera with push pull rod located above binocular vertical control knob. 3. Start StereoInvestigator. Select Imag- ing > Live Image (Ctrl-L) 4. Click with mouse inside image frame to set reference point and to improve performance of mouse when hovering over the display area. - Page 9 Acquire Images Objective List All require 0.17mm coverslip unless noted With StereoInvestigator Spatial Calibration: 2 5x Plan-Neofluar Olympus Microfire color CCD Acquire Image Stacks (Z-Series). 2.5x Bin 1 ..2.8 mm/pixel NA .
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Page 10: Saving Files
Saving files 1. Save to Local Machine. --Default You may save files to Workspace (E:\Workspace\”username”). A folder was created there at login for each user. 2. Before leaving, save to Network Share -- “username” on ‘ITG File Server (zeus.itg. uiuc.edu)’(Z:) Most users opt to save the data to their network share on our central server (named Zeus). - Page 11 References Chroma Filter Sets for AxioImager A1 Calibration Standards Calibration Grid Slide from Micro Brightfield: Excitation Wheel Filter Cubes/Dichroics http://www.mbfbioscience.com (1) Blank (1) Transmitted cube with polarizer (2) Dual Cy2/Cy3 (2) Cy2/Cy3 filter cube for viewing Fluorescent Reference Slides from Micros-...