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Marsico Hall Microscopy Fellowship (MHMF.ORG)

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Using the LeicaSP2-AOBS Confocal Microscope

** If you have ANY questions, please contact a facility director **

1. Powering Up - assuming entire system is off

        1.1  Assess status of the system

        1.2  Turn on mercury arc lamp, microscope stand and scanner

        1.3  Turn on the required laser(s)
                      **Only turn on laser which are required for the dyes in your samples**
                      Properly power down any laser which is not required (see section 8 below)

    1.4  Log on to WindowsXP

3. Starting the Confocal Scanning Program

2.  Viewing the Sample

2.1  Setting the stage (general information for fluorescence or transmitted light viewing)

        For fluorescence only go to section 2.4 (skip sections 2.2 & 2.3)

        2.2  Transmitted light (specific setup instructions)

        2.2a For Imaging transmitted light, set up Kohler illumination
               
Kohler illumination is not important for fluorescence or confocal imaging. 
                However, if simultaneous confocal and transmitted scanning are done, Kohler illumination should be set up

        2.3  DIC/Nomarski - (if required)

        2.4  Fluorescence

4. Scanning the sample

 

5. Saving Images (brief description below)

Return to looking through the microscope by pressing the MicCtrl button and choosing Visual


6. Shutting Down - ALL users MUST follow these shut down procedures carefully


7. Confocal Scanning Tasks

8. Image Processing