Zeiss LSM 880

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Skip to Instrument Specifications below.

 

Startup:

      

DO NOT touch any switches except the the ones in these instructions.

If you are only using the computer, only turn on the switches in step 3; leave all the other ones off.

  1. SIGN IN LOG BOOK
  2. SWITCH 1
    MAIN SWITCH

    Turns on: DEFO, HXP, incubation
  3. SWITCH 2
    Red Switch on Power Strip

    Sends power to the PC (computer) and monitor.
    • Press the power button on the front of the CONFOCAL computer on the floor to the right.
    • Only turn on the confocal computer.
    • You can use Zen Blue software for data processing instead of Zen Black (which runs the hardware).
      (Please ask for more training if you want to be shown this.)
    • If you are only doing data processing, stop here.
    • DO NOT TURN ON THE FLIM COMPUTER UNLESS YOU ARE DOING FLIM. If you don't know what FLIM is, you are not using it.
  4. SWITCH 3
    SYSTEMS/PC
    (label not descriptively correct...)
    Turns on additional confocal components.

    STOP. Wait for computer to completely boot up before next step.
     
  5. SWITCH 4
    COMPONENTS

    Turns on: Lasers, scan head, real time PC, etc.
  6. STOP. The microscope control touch pad must be on before you continue.
    If the microscope touch pad does not light up, try pressing the power button at the back left of the microscope stand.
  7. Log in to computer account. (Password taped to screen.)
  8. Run Zen software.
  9. Immediately go in to Lasers window in Acquitiion tab and turn on the lasers you will be using. This will save time by allowing the lasers to power up fully while you find your sample, load settings, etc.
  10. Optional: Incubator may be set by the touchpad on the table.
    Home > Microscope > Incubation
    For Airy Scan, the Incubator should be on at 30 degrees C and make sure the doors and other covers on the incubator box are closed.

General Confocal Best practices:

 

Oil and Water lenses

Use special 30 degrees oil for incubation only.

Use 1.518 oil for RT.

Use Immersol W or GenTeal for water lenses.
DO NOT LEAVE IMMERSOL W BOTTLE ON THE TABLE.
AT THE END OF YOUR IMAGING SESSION, MAKE SURE YOU RETURN IT TO THE SHELVES at the back of the room.

 

Saving Files

All files should be stored in one of these two locations.
   D:\User_Data\
   D:\SWAP\
Files left on the desktop or drive C: will be deleted.

.czi always.

Move data to your lab's shared server space.

 

Detectors

CH1 and CH2 GaAsP detectors require 500V gain minimum.

For BiG use plate, not mirror. Filter blocks installed manually in side. Blocks for red/green and CFP/YFP.

 

Software Help

Click on ? button in upper right and then click on thing on screen want help with.

 

Tiling

First do 128X128 large area fast overview tile scan.
Put in new container.
Click stage (or use positions to set multiple Z)

12% overlap.

Zen Blue for post processing stitching.

 

Airyscan

When using Airyscan, do not change the size of the image or the zoom manually. Use the Optimize button to choose the correct settings.
DO NOT THINK, just click Optimize.
The pinhole should also be open to the right of the red bar in the pinhole slider.


Power down if no one using system immediately after you:
Turn off Ar laser in software.
Quit software.
Transfer files to server.
Shut down computer (menu from button lower left of screen or from Ctrl-Alt-Del).
After computer shut down, power switches off in reverse order as startup.
DO NOT turn any key.

If you did not touch the switch on startup, do not touch it on shutdown.

 

Power down if yes someone using system immediately after you:
Transfer files to server.
Do not turn off Ar laser in software.
Quit software.
Log out of computer but do not shut down (menu from button lower left of screen or from Ctrl-Alt-Del).
Log in quickly and immediately start Zen again.

 

 

Specifications:

Axio Observer.Z1, Zeiss full environmental chamber with temp/CO2

Lasers at 405, 440, 458 (weak), 488, 514, 561, 594, 633 nm.

Autofocus system using 780(?) nm laser. Especially useful for multifield timelapse imaging. Keeps focus locked for hours-days.

Detectors that may be used simultanously for standard imaging:
 PMT "Ch1"
 34 spectral channel GaAsP which may be defined as multiple channels or binned as desired "ChS1" to "ChSn"
 PMT "Ch2"
 T-PMT (transmitted light; align condenser to use)

Flip the mirror for:

Detectors that may be used simultanously for FCS, FLIM, low light with manually installed bandpass filters:
 GaAsP1
 GaAsP2

AiryScan "ChA"

Example time domain FLIM using 440 PicoQuent laser.


 

Maintenance issues:

Adding oil to compressor.

MIC-SYSTEM Ser Nr 2802000293
Scanmodul 34Ch-G 24V= 1997-779
and
Rack LSM 880
Ser Nr 2817000414

 

 

 

 

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comments, questions, suggestions: Michael.Cammer@med.nyu.edu