Center for Cell Death, Injury and Regeneration

510

Before using these instructions make sure you have had at least one training sessions on using the system.

Zeiss LSM 510 META operating instructions:
1. Turn on the mercury arc lamp the first if you intend to use it. Wait for 2 minutes for  the lamp to warm up before turning on any other piece of equipment. Switch off the arc lamp only after all the other equipment is turned off.  

2. Switch on the computer and system component button on the box besides the computer and switch on the computer.

3. After computer boots up logon with
Username: administrator
Password: zeiss

4. The computer will prompt you to enter your username and password that was created during the training session. This account is used for tracking users and their usage time for billing.

5. Once logged on the computer, launch the LSM 510 Meta software. Start software in Expert mode.

6. Select laser from the 510 control panel and switch on the laser lines that you intend to use and keep the lasers running for 30 minutes before starting an experiment. Do not turn lasers on and off during an experiment. The microscope has the following laser lines available:
A. 458, 477, 488 and 514 nm Argon Laser
B. 543 nm He/Ne laser
C. 633 nm He/Ne laser

7. Click on Config and select single or multitrack.Then load the configuration that you intend to use for the experiment using the Config button. Configurations will be created by the facility managers and stored for subsequent use.

8. Select the objective lens that you intend to use for the experiment. Make sure you use the appropriate immersion medium. Do not use the wrong medium as it may damage the lenses. Lenses currently available on microscope:
A. 10X dry lens
B. 63X oil immersion lens (use Zeiss 518F immersion oil)
C. 40X oil immersion lens (use Zeiss 518F immersion oil) 

9. Switch to VIS mode on LSM software

10. Mount the specimen on the microscope stage and bring it in focus using transmitted or fluorescent light while looking through the eye-piece.

11. Once specimen is focused switch to LSM mode.

12. Launch the scan menu and select frame size, scan speed and type of scan.      

14. Start an experiment with an laser intensity of 0.05%. Use fast scan to bring specimen into fine focus. While doing a fast scan check palette on the iumage window and select range indicator. Looking at the image adjust detector gain, amplifier offset and laser intensity till you get an image with a few red and blue pixels. 

15. Switch to slower scan to collect images. Use scan averaging if necessary to improve S/N ratio.

16. Save all your images to the folder you have created under D:\Userdata\Yourname. Before you start saving images you will be asked to create a database. Create the database under the same folder. Also the software saves all images in default LSM format. Use the export function to save them in other formats like TIFF, JPEG. Ask the managers to show you how to use this.

17. Once you finish your experiment shut down all the lasers.

18. Wait for 5 minutes for lasers to cool down.

19. Remove your specimen from the stage and clean the lens using Zeiss cleaning solution while you wait for the lasers to cool down. Also save all your data to a CD, DVD or USB memory. The facility is not responsible for storing images.

20. After the lasers have cooled exit the LSM software shut down the computer and switch the system and components button to off.

21. Turn off the arc lamp at the end.  

22. Cover the microscope and tidy up the workplace.

 
 
 

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