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Objectives

  • 4x
  • 10x (pixel size 0.655 um)
  • 20x

Filter specifications

FilterWavelengthsShading correction plate
DAPI

Excitation: 377/50 nm

Emission: 447/60 nm

Dichroic: 409 nm

Green

Pink

GFP

Excitation: 472/30 nm

Emission: 520/35 nm

Dichroic: 495 nm

Pink

Red

Texas Red

Excitation: 562/40 nm

Emission: 624/40 nm

Dichroic: 593 nm

Red
Cy5

Excitation: 628/40 nm

Emission: 692/40 nm

Dichroic: 660 nm

Red


Plate calibration

Plate calibration is done in two steps:

  1. Measure variation of plate bottom thickness
    • requires an empty plate
  2. Measure required exposure time for autofocus laser
    • autofocus is based on the reflection of the laser from the plate and well bottom
    • reflection from the well bottom depends on the refractive index of the mounting media
    • requires a plate with relevant sample 

The system has now these calibrated settings:

ManufacturerPlate modelObjective and mounting
4titudeVisionPlate 96-well


Costar3596 96-well10x PBS
Falcon353072 96-well10x, water
GreinerCellstar 655 180 96-well20x PBS
NuncOptical 96-well10x, air
PerkinElmerCellCarrier Ultra 96-well
PerkinElmerViewPlate 96-well10x, PBS
PerkinElmerCellCarrier Ultra 384-well10x, PBS



If you use any other plates, you need to bring two plates for the training:

  1. empty plate
  2. a plate with sample (and media in the wells with no sample)


Starting the system

  1. Power on the instrument.
  2. Start computer.
  3. Log in with account "Nano"
  4. Mount L: disk
    1. Double-click "lmu-L-disk" on the Desktop.
    2. In the command window, write your university username in the format ATKK\username, press Enter.
    3. In the command window, write your university password, press Enter.
  5. Start MetaXpress
    1. Double-click "MX" icon on the Desktop.
    2. Select your data source (named after your university username).

Troubleshooting network problems

  1. Check with LMU staff that you have correctly mounted the L-disk. 
    1. If L-disk is not available, use the local F disk (selector is in the "Run" tab).
  2. Check if you can connect to your usual data source
    1.  If your usual data source doesn't work, select data source zzz_<username>_local, which uses the local database. 


Shutting down the system

  1. Close software
  2. Disconnect L: disk (to delete login credentials)
    1. Start Menu → Computer
    2. Right-click Network Location lmu (L:)
    3. Select Disconnect
  3. Shut down computer
  4. Power off the instrument





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