Instructions for zeiss axioimager a1 microscope for stereology basic operation instructions as of 10/1/2009 the stereology workstation by jon ekman.
Start-up 1. Turn master switch on belkin surge- master. This will turn on the ludl stage controller, microscope stand and the olympus microfire color ccd camera. 2. Turn on x-cite 120 hg lamp if needed. Note: the x-cite 120 must stay on a minimum of 20 minutes. It also requires 20 minutes to cool do...
Align optical components koehler illumination procedure: transmitted light 1. Focus on specimen with the objective to be used for data collection. 2. Close down the luminous field dia- phragm (1) while viewing. 3. Raise or lower condenser vertical control slightly (2) until luminous field diaphragm ...
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...
Stereology with the optical fractionator workflow tissue preparation the optical fractionator requires fairly thick tissue. Thicker tissue allows one to distinguish between cells that are on top of each other while focusing through the tissue making it easier to see the cells clearly as they start t...
Optical fractionator workflow continued... 100x-oil). Objective choice will depend on tissue type and staining. Note: objec- tives with magnification lower than 40x will not give you enough depth of view to count accurately especially when cells are close to each other in z. Step 5: measure section ...
Do not save each section as a new file. Use the workflow to add new sections, as described above, until all the sections in an animal are traced and counted. Note: one animal = one file. And past the bottom of the tissue until it is completely out of focus and then slowly focus back up until somethi...
Start-up 1 . . . . . . . . . Turn on master switch 2 . . . . . . . . . . . Log on to computer 3 . . . . . . . . . . . . . Load software optional . . . . .Turn on x-cite hg lamp align optical components 1 . . . . . . . . . . .Koehler illumination calibrate system 1 . . . . . . . . . . . . . Grid tune...
Acquire images objective list all require 0.17mm coverslip unless noted 2 5x plan-neofluar 1 na . . . . . . . . . . . . . . . . . 0.075 wd(mm) . . . . . . . . . . . . . . . 9.5 depth of field (560nm) . . . . . . 139mm part # . . . . . . . . . . . . . . . 440310 5x plan-neofluar 1 na . . . . . . . . ...
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...
References calibration standards calibration grid slide from micro brightfield: http://www.Mbfbioscience.Com fluorescent reference slides from micros- copy education: http://www.Microscopyeducation.Com olympus http://www.Olympusamerica.Com/seg_sec- tion/seg_home.Asp -- olympus microfire color ccd • ...
