MolecularDevices Nano

Last modified by Harri Tapio Jäälinoja on 2024/02/12 16:52

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:

  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:

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:

  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. You can also use the shared account MDCStore (see credentials), 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 (Lemoticon_smile
    3. Select Disconnect
  3. Shut down computer
  4. Power off the instrument