MolecularDevices Nano
- Nikon Objectives
- Filter specifications
- Plate calibration
- Starting the system
- Troubleshooting network problems
- Shutting down the system
Nikon Objectives
- 4x/0.2 Plan APO Lamda, WD 20mm (pixel size 1.64 um)
- 10x/0.3 Plan Fluor, WD 16mm (pixel size 0.655 um)
- 20x/0.45 S Plan Fluor ELWD, WD 8.2-6.9mm (pixel size 0.328 um)
Filter specifications
Filter | Wavelengths | Shading 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 |
https://www.fpbase.org/microscope/4EM62MNAT3Tdh4jaa7GVNS/
Plate calibration
Plate calibration is done in two steps:
- Measure variation of plate bottom thickness
- requires an empty plate
- 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:
Manufacturer | Plate model | Objective and mounting |
---|---|---|
4titude | VisionPlate 96-well |
|
Costar | 3596 96-well | 10x PBS |
Falcon | 353072 96-well | 10x, water |
Greiner | Cellstar 655 180 96-well | 20x PBS |
Nunc | Optical 96-well | 10x, air |
PerkinElmer | CellCarrier Ultra 96-well |
|
PerkinElmer | ViewPlate 96-well | 10x, PBS |
PerkinElmer | CellCarrier Ultra 384-well | 10x, PBS |
|
|
|
If you use any other plates, you need to bring two plates for the training:
- empty plate
- a plate with sample (and media in the wells with no sample)
Starting the system
- Power on the instrument.
- Start computer.
- Log in with account "Nano"
- Mount L: disk
- Double-click "lmu-L-disk" on the Desktop.
- In the command window, write your university username in the format ATKK\username, press Enter.
- In the command window, write your university password, press Enter.
- Start MetaXpress
- Double-click "MX" icon on the Desktop.
- Select your data source (named after your university username).
Troubleshooting network problems
- Check with LMU staff that you have correctly mounted the L-disk.
- If L-disk is not available, use the local F disk (selector is in the "Run" tab).
- Check if you can connect to your usual data source
- You can also use the shared account MDCStore (see credentials), which uses the local database.
Shutting down the system
- Close software
- Disconnect L: disk (to delete login credentials)
- Start Menu → Computer
- Right-click Network Location lmu (L
- Select Disconnect
- Shut down computer
- Power off the instrument