Using the facility

Equipment available at the facility

Users of the LCI facility have access to the following equipment:
- 4 Nikon confocals (Orion, Tweety, Dragon, Pegasus)
- 1 Zeiss single and multiphoton confocal (Zeus)
- 2 spinning disk confocals (Victor, Tweety)
- 4 widefield microscopes (Victor, Orion, Tweety, MMI)
- 2 TIRF microscopes (Orion, Victor)
- STORM super resolution microscope (Orion)
- High content screening modules (Dragon, Tweety, Orion, Pegasus, Victor, MMI)
- 1 micropatterning and micromanufacturing device (Tweety)
- 1 laser capture microdissection and cell picking system (MMI)
- Slide scanner (MMI)
- 4 offline computers with several analysis and deconvolution software

Some of our confocals are equipped with fast resonant scanners, spectral detectors and FLIM (Fluorescence Lifetime Imaging)
Most of our microscopes have a large incubator for long term live cell imaging.

Click on the individual microscope names on the right menu to find out details about each system.

 

Which microscope should I use?

If you want to image fast, you probably need a widefield system.
Confocals, point scanning or spinning disk, are ideal to image samples that are thicker than 10 um.
For very thick samples, the multiphoton might be needed.
TIRF systems are used to image the area of the cell close to the coverslip.
If the resolution needed is higher than 200 um, you will need super resolution.
FLIM allows you to image protein-protein interaction using FRET.
With our micropatterning system, you can print proteins at the bottom of a dish or on a gel and study their interaction with cells. We can also make custom designed microwells.
Use our laser capture microdissection system to cut part of a tissue and pick it for RNA analysis or Mass Spectrometry.
The analysis software on our workstation can analyse multidimensional image files: xy, z, multicolour, time...

 

Access and Booking

Fees

Fees at the LCI Unit

The Live Cell Imaging facility is open to all researchers. Prices are listed below.

Users pay an entry fee once. This includes training as well as help with sample preparation, experimental design and image analysis. Additionally, users pay the usage fee, every year, 3 or 6 months depending on the length of their project. There is no additional fee per hour.

The entry fee for the Nikon confocals gives access to 4 confocals (Tweety, Pegasus, Dragon and Orion).
The entry fee for the Nikon widefield systems gives access to 3 widefield systems (Orion, Victor and Tweety), 2 TIRF systems (Orion and Victor), STORM (Orion) and 2 spinning disk confocals (Victor and Tweety).

The entry fee for the Zeiss confocal gives access to 1 confocal with a multiphoton option (Zeus).

Users of the microscopes have access to all analysis software without any additional entry fee (Imaris, NIS and MatLab).

 

1. Entry fee

Microscopes and analysis software, first time using the LCI facility 12.000 SEK
Microscopes and analysis software, already user of the LCI facility 9.000 SEK
Analysis software only (non-microscope users) 5.000 SEK
MMI laser microdissection and cell picker 2.000 SEK
PRIMO micropatterning (including reagents for 5 dishes) 2.700 SEK
PRIMO micromanufacturing (including practice glue) 2.000 SEK

2. Usage fee

  12 months 6 months 3 months   1 month 2 weeks
Microscopes and analysis software 16.500 SEK 10.000 SEK 6.500 SEK   N/A N/A
Analysis software only 5.000 SEK 3.500 SEK 2.000 SEK   N/A N/A
Primo or MMI 12.000 SEK 9.000 SEK 6.000 SEK   2.500 SEK 1.500 SEK

For non academic users, the yearly fee is double.

 

Registration and booking

Booking at LCI Unit

Fee form confocal and widefield

Fee form MMI and PRIMO

Booking (log-in required)

 

Laser safety

Laser safety at the LCI unit

Lasers at the LCI facility microscopes fire visible or invisible light. Invisible lasers are the 405 nm laser and the multiphoton laser.
Never move your sample when the system is scanning, even if you do not see any light! Otherwise the sample may act as a mirror.

 

Guidelines

Guidelines LCI Unit

The LCI facility guidelines must be followed by all users.

 

Tweety

 

 

This system can be used as:
- Single point scanning confocal
- Spinning disk confocal
- Ultra fast widefield
- Micropatterning (printing proteins on a dish) and micromanufacturing (making microwells)

Equipment specifications
- Nikon
- Inverted
- Full size incubator for life cell imaging
- Fully motorized including Z piezo 100 um
- High content screening JOBS module
- Andor camera Zyla 4.2+. 100 fps at 2048x2048 pixels. Pixel size 6.45 um. QE 82%
- External filter wheel and LED for fast imaging
- Alvéole Primo micropatterning and micromanufacturing device

Filters on the widefield side

Cube name     Excitation Filters Dichroic Mirrors Emission Filters
Blue
Green
Red
Far Red
    387/11
472/30
562/40
628/40
409LP
513LP
601LP
660LP

447/60
536/40
624/40
692/40

Filters on the confocal side

Lasers Dichroic Mirrors Emission Filters

405 diode
488 diode
561 diode
640 diode

405/488
405/488/561
405/488/561/640
BS 20/80

450/50
525/50
595/50
700/75

Pegasus

 

This system can be used as:
- Single point scanning confocal

Equipment specifications
- Nikon
- Inverted
- Full size incubator for life cell imaging
- Fully motorized including Z piezo 100 um
- High content screening JOBS module
- Resonant scanner for fast imaging
- 2x GaAsP detectors (green and red)

Lasers Dichroic Mirrors Emission Filters

405 diode
457/488/514 Argon Ion
561 diode
640 diode

405/488
405/488/561
405/488/561/640
405-457/514
457/514/561
BS 20/80

450/50
482/35
525/50
540/30
595/50
700/75

Dragon

 

  

This system can be used as:
- Single point scanning confocal

Equipment specifications
- Nikon
- Inverted
- Full size incubator for life cell imaging
- Fully motorized
- High content screening JOBS module
- Resonant scanner for fast imaging
- 2x GaAsP detectors (green and red)
- 32-channel spectral detector
- La Vision BioTech FLIM (Fluorescence Live time Microscopy)

Lasers Dichroic Mirrors Emission Filters

405 diode
457/488/514 Argon Ion
561 diode
640 diode

405/488
405/488/561
405/488/561/640
405-457/514
457/514/561
BS 20/80

450/50
482/35
525/50
540/30
595/50
700/75

Orion

 

This system can be used as:
- Single point scanning confocal
- Widefield
- TIRF
- STORM super resolution

Equipment specifications
- Nikon
- Inverted
- Full size incubator for life cell imaging
- Fully motorized including Z piezo 100 um
- High content screening JOBS module
- 32-channel spectral detector
- Andor EM-CCD camera, 60 fps at 512x512 pixels. Pixel size 16 um. QE 95%

 

Filters on the widefield side

Cube Excitation Filters   Dichroic Filters Emission Filters

Blue
Green
Red
Far Red

387/11
472/30
562/40
620/60
  409LP
495LP
593LP
660LP
447/60
520/35
624/40
700/75

Filters on the TIRF/STORM side

Cube Excitation Filters Dichroic Filters Emission Filters
Dual TIRF 480-495
555-565
495-545
565LP
500-545
570LP
Quad TIRF 385-418
485-495
555-570
635-660
420-480
495-555
575-630
670LP
422-480
500-550
580-625
675LP
STORM   405/5
488/5
561/5
647/5
670-750

Filters on the confocal side

Lasers Dichroic Mirrors Emission Filters

405 diode
457/488/514 Argon Ion
561 diode
640 diode (300 mWatt)

405/488
405/488/561
405/488/561/640
405-457/514
457/514/561
BS 20/80

450/50
482/35
525/50
540/30
595/50
700/75

Victor

 

This system can be used as:
- Spinning disk confocal
- Ultra fast widefield
- TIRF

Equipment specifications
- Nikon
- Inverted
- Full size incubator for life cell imaging
- Fully motorized including Z piezo 250 um
- High content screening JOBS module
- Widefield: Andor camera Zyla 4.2+. 100 fps at 2048x2048 pixels. Pixel size 6.45 um. QE 82%
- External filter wheel and SpectraX LED for fast imaging
- Spinning disk: Andor EM-CCD camera, 60 fps at 512x512 pixels. Pixel size 16 um. QE 95%
- Spinning disk Yokagawa CSU-X1, 10,000 rpm

 

Filters on the spinning disk confocal side

Lasers Quad Dichroic Mirror Emission Filters

405 nm diode
488 nm diode
561 nm diode
640 nm diode

425-480
503-555
580-628
675LP

447/60
512/30 ; 630/100
525/30
525 600 690
607/36
685/40

Filters on the widefield/TIRF side

LEDs Dichroic Mirrors Emission Filters
395/25 nm
440/20 nm
475/28 nm
510/25 nm
550/15 nm (or 575/25)
631/28 nm
425-480
503-555
580-628
675LP
447/60
483/32
500LP
525/30
542/27
607/36
630/92
685/40

Zeus

 


 

This system can be used as:
- Single point scanning confocal
- Multiphoton point scanning confocal

Equipment specifications
- Zeiss LSM710-NLO
- Upright
- Full size incubator for life cell imaging
- Fully motorized
- 34-channel spectral detector
- GaAsP detector
- 4 NDDs (2 trans and 2 epi)

Lasers
- 405 diode
- 457/488/514 Argon
- 561 diode
- 633 diode

 

MMI Laser microdissection and cell picker

 

This system can be used as:
- Widefield
- Slide scanner
- Laser microdissection for RNA and protein analysis of part of a tissue
- Cell picker and sorter

Equipment specifications
- Nikon
- Inverted
- Fully motorized
- High content screening JOBS module
- Nikon colour camera
- Andor EM-CCD camera, 60 fps at 512x512 pixels. Pixel size 16 um. QE 95%
- MMI CellCut and CellEctor

Filters for widefield

Cube Excitation Filters Dichroic Mirrors Emission Filters
Blue 360-370 400LP 435-485
Green 450-490 495LP 500-530
Red 532-557 565LP 570-640
Far red 608-648 660LP 672-712

Objectives

Objectives that are permanently available on each of our NIKON microscopes

Magnification Type n NA WD mm
10x Dry 1 0.45 4
20x Dry 1 0.75 1
60x Oil 1.55 1.40 0.13

Objectives that are rotating between our NIKON microscopes

Magnification Type n NA WD mm
40x Dry 1 0.95 0.14-0.25 (ring)
40x Oil 1.52 1.30 0.22 (ring)
60x (TIRF) Oil 1.52 1.49 0.12 (ring)
60x Water 1.33 1.20 0.28-0.31(ring)
100x (TIRF) Oil 1.52 1.49 0.12 (ring)

Objectives available on Zeus

Magnification Type n NA WD mm
5x Dry 1 0.16 18.5
20x Dry 1 0.6 1.7
20x Dipping 1.33 1.00 1.8
40x Water 1,33 1.10 0.62
63x Water 1.33 1.20 0.28

Objectives available on MMI

Magnification Type n NA WD mm
4x Dry 1 0.13 16.5
10x Dry 1 0.3 15.2
20x Dry 1 0.45 (ring) 8.2-6.9
40x Dry 1 0.6 (ring) 3.6-2.8
60x Dry 1 0.75 (ring) 2.62-1.8
20x Dry   0.75 1

Software

We have 4 offline stations at the LCI Facility equipped with the following softwares:

- NIS Element: offline software for the Nikon systems
- Zeiss Zen: offline software for Zeus
- Imspector Pro: offline software for FLIM
- Imaris: powerful 5D rendering and statistical analysis software
- Cell Profiler: 4D (no z) rendering and statistical analysis freeware
- ImageJ/Fiji: image processing and visualizing freeware
- XUV Tools: 3D image alignment freeware
- MatLab: custom-made analysis

Microscopy