MolecularDevices Micro
Using Micro
Starting the system
- Power on the instrument.
- Start computer.
- Log on with your Windows account
- Mount L: disk
- Double-click "lmu_active1.bat" 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 "20pa20ph10pa40wi IN Carta" icon on the Desktop.
Select your data source (university username).
Known well-plates and 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 |
|---|---|
| Akita | 96-well |
Greiner | Cellstar 655180 96-well F-bottom |
| Greiner | Cellstar 657160 6-well |
| Greiner | Cellstar 665180 12-well |
| Greiner | Cellstar 662160 24-well |
| Greiner | Cellstar 677180 48-well |
| Ibidi | 8-well slide (with slide holder) |
Ibidi | uPlate 24-well |
| Ibidi | uPlate 96-well |
Revvity | PhenoPlate 96-well |
PerkinElmer | ViewPlate 96-well |
Thermo Scientific | 442404 MaxiSorp 96-well |
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)
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
The instrument power can be left always on. Here are the steps for the other components:
- Close the software.
- Check OpenIRIS for the next booking.
- There is a booking within 2h: Log out.
- There is no booking with 2h: Log out, shut down computer.
- Does the next user within 24h need the system warmed up?
- Yes: Power on heating.
- If you used CO2, close CO2 bottle valve and regulator output valve.
Technical details
Nikon Objectives
| Objective | NA | Resolution 0.61 x 509 nm / NA | Pixel size | Working distance |
|---|---|---|---|---|
| 4X CFI Plan Apochromat Lambda D | 0.2 | 1.55 um | 20.0 mm | |
| 10X Plan Apo Lambda | 0.45 | 690 nm | 676 nm | 16 mm |
| 20X Plan Apo Lambda | 0.8 | 388 nm | 349 nm | 0.8 mm |
| 20X Ph1 S Plan Fluor ELWD ADM | 0.45 | 690 nm | 340 nm | 6.9-8.2 mm |
| 40X Water Apo LambdaS LWD | 1.15 | 270 nm | 170 nm |
The instrument only has room for four objectives at a time. There are two hardware configurations where either the 4x or 20x/0.45 objective is left out:
- 20pa20ph10pa40wi
- 20pa4pa10pa40wi
Light source (LED)
Filter specifications
Excitation filters are built in the Lumencor light source (see ).
| Fluorophores | Wavelength (nm) | Power (mW) |
|---|---|---|
DAPI | 377/54 | 200 |
| FITC | 475/28 | 200 |
| TRITC | 543/27 | 300 |
| TexasRed | 560/32 | 200 |
| Cy5 | 631/28 | 155 |
Filter specifications according to MolecularDevices documentation.
Filter | Wavelengths |
|---|---|
DAPI | Excitation: 377/54 nm Emission: 447/60 nm Dichroic: 409 nm |
FITC | Excitation: 475/28 nm Emission: 536/40 nm Dichroic: 506 nm |
| Cy3 (TRITC) | Excitation: 543/27 nm Emission: 593/40 nm Dichroic: 562 nm |
Texas Red | Excitation: 560/32 nm Emission: 624/40 nm Dichroic: 593 nm |
Cy5 | Excitation: 631/28 nm Emission: 692/40 nm Dichroic: 660 nm |
Camera
4.2 Mpixel sCMOS camera.
- 2160 x 2160 pixels
- 16 bit
- 82% QE
- 40 fps
- TIFF format