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6) Now you can start the software LAS-AF and insert your sample into

the sample holder. As the start-up routine moves the stage, it is not
recommended to focus into your sample until after the software has
fully started.

7) Choose either machine – with pump or machine – without pump

according to your needs. Usually the pump is ever only used with wellplates
when 63x water immersion objective is needed. Start the software from the
OK button.

Summary of Contents for SP5 II

Page 1: ...Quick Manual How to start and shut down Leica SP 5 II HCA ...

Page 2: ...ou need to change it Be careful not to drop the small screws used to fasten the sample holder into the motorized stage 4 Turn on the microscope A the scanner B the laser power C and turn the safety key to On 1 position 5 Turn on the fluorescent lamp if you plan to see fluorescent light through the binoculars The intensity of the light can be adjusted from the round knob but this is usually unneces...

Page 3: ... the stage it is not recommended to focus into your sample until after the software has fully started 7 Choose either machine with pump or machine without pump according to your needs Usually the pump is ever only used with wellplates when 63x water immersion objective is needed Start the software from the OK button ...

Page 4: ...m bottom and Mattek dishes have focus about 1mm above that 9 Once the software has started chose Configuration Lasers and turn on all the lasers you will use If you are using Argon laser adjust its power to 20 from the slider If you want to save your data in other than 8 bit format recommended if you plan to quantify your data go to Configuration Settings and choose the 12 bit option ...

Page 5: ...med Leica settings Alternatively sequential settings can be loaded from sequential settings 11 Adjust the detection scheme open the shutters as needed and adjust the laser powers as needed Remember to adjust the beam splitter BS according to your needs Using CO2 1 To use the gas exchanger you must open the air and CO2 valves as shown below Also remember to turn on the gas mixer itself Brick the re...

Page 6: ...se LAS AF software 5 Remove your sample and carefully clean the objective s 6 If no one has a reservation within two hours of your reservation turn off the PC Microscope and Scanner Power switches Disable the lasers by turning the key to Off 0 position Wait until Argon laser cooler has stopped before turning Laser Power switch off 7 Check if the next person after you within 24 hours needs the micr...

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