Microscopes at LCI

The Live Cell Imaging Core Facility has grown to include 8 high-end microscopes. Our microscopes allow researchers to perform conventional and high throughput confocal and widefield microscopy, fast confocal microscopy (resonance scanner and spinning disk), Fluorescence Life Time Imaging (FLIM), multiphoton microscopy, spectral imaging, super resolution microscopy (TIRF and STORM) and light sheet microscopy.

  • 1 Zeiss single and multiphoton confocal (Zeus)
  • 2 Nikon single point confocals (Dragon, Pegasus) with fast resonant scanners, and spectral detectors.
  • 3 Nikon multipoint confocals (spinning disk) (Victor, Orion and Tweety)
  • 5 Nikon widefield (Victor, Orion, Tweety, Pegasus, Minerva)
  • 2 Nikon TIRF (Pegasus, Victor)
  • 1 Nikon STORM super resolution (Pegasus)
  • 1 Nikon DeepSIM super resolution (Orion)
  • 1 M-Squared Light Sheet (Aurora)
  • Nikon JOBS High content screening module (Dragon, Tweety, Orion, Pegasus, Victor)
  • 1 Primo (Alveole) micropatterning and micromanufacturing device (Tweety)
  • 1 MMI laser capture microdissection and cell picking system (Minerva)
  • 1 MMI Slide scanner with a colour camera (Minerva)
  • Imaris, NIS-Element, Zen, Fiji, Cell Profiler, Deconvolution, ImSpector, MatLab, Python, CTFire, Omero

Most of our microscopes have a large incubator for long term live cell imaging.

Tweety

This system can be used as

  • Spinning disk CREST v3
  • Ultra fast widefield with large field of view 
  • 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
  • 2 very sensitive cameras from Photometrics: Prime 95B sCMOS, 95% QE, 11um pixels for best sensitivity, 1608x1608 pixels and Kinetix sCMOS, >95% QE, 6.5 um for best resolution, 2720x2720 pixels   
  • Alvéole Primo micropatterning and micromanufacturing device

Filters for Widefield and Spinning disk side

  • Lasers:  405, 446, 477, 520, 546, 638, 749 (nm)
  • Dichroic mirrors: 441/30; 511/26; 593/37; 684/34; 817/66 (nm)
  • Dichroic mirrors: 484/22; 561/30
  • Dichroic mirror: 562LP
    • Pentaband: 441/30; 511/26; 593/37; 684/34; 817/66 (nm)
    • Dual band: 484/20; 561/30 (nm)
    • Single bands: 438/24; 485/20; 511/20; 560/25; 595/31; 685/40; 561LP (nm); 819/44nm

For more information on

Tweety widefield and spinning disk: Semrock searchlight

Pegasus

This system can be used as

  • Single point scanning confocal
  • Resonant scanner for fast imaging
  • 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
  • 2x GaAsP detectors (green and red)
  • 32-channel spectral detector
  • Andor EM-CCD camera, 60 fps at 512x512 pixels. Pixel size 16 um. QE 95%

Filters on the widefield side

  • Excitation filters:  387/11; 472/30; 562/40; 620/60 (nm)
  • Dichroic mirrors: 409LP; 495LP; 593LP; 660LP (nm)
  • Emission filters:  447/60; 520/35; 624/40; 700/75 (nm)

Filters, lasers and dichroic mirrors for the confocal

  • Lasers:  405 (diode), 457/488/514 (Argon Ion) 561 (diode); 647 (diode) (nm)
  • Dichroic mirrors: 405/488 ; 405/488/561; 405/488/561/640; 405-457/514 ; 457/514/561; BS20/80 (nm)
  • Emission filters: 450/50; 482/35; 525/50; 540/30; 595/50; 700/75 (nm)

Filters, lasers and dichroic mirrors

  • Lasers:  405 (diode), 457/488/514 (Argon Ion) 561 (diode); 640 (diode) (nm)
  • Dichroic mirrors: 405/488; 405/488/561; 405/488/561/640; 405-457/514; 457/514/561; BS20/80 (nm)
  • Emission filters: 450/50;482/35; 525/50; 540/30; 595/50; 700/75 (nm)

Filter cubes for STORM

  • STORM cube: 405; 488; 561; 647,  (5 nm width). Emission filter: 705/70 nm.

Filter cubes for TIRF

  • Dual TIRF: Excitation filters:  488/561 (5 nm width); DM: 507/24; 565LP (emission) (nm); Emission filters: 522/45; 570LP (nm)
  • Quad TIRF: Excitation filters; 405; 488; 561; 647, (5 nm width); DM: 450/60; 525/60; 597/55; 607/55: 670LP (emission); Emission Filters: 451/29; 525/50; 607/18; 675LP (nm)

For more information on

Pegasus widefield: Semrock searchlight
Pegasus single point confocal: Semrock searchlight

Dragon

This system can be used as

  • Single point scanning confocal
  • Resonant scanner for fast imaging with large field of view (1024x1024 pixels)
  • Wide field with colour and monochrome imaging

Equipment specifications

  • Nikon
  • Inverted
  • Full size incubator for life cell imaging
  • Fully motorized
  • High content screening JOBS module
  • 2x GaAsP detectors (green and red)
  • 32-channel spectral detector
  • Digital Sight 10 CMOS colour camera from Nikon, 25 mm FOV, 5.97 pixel size, for best resolution 6000 x 3984 pixels

Filters, lasers and dichroic mirrors

  • Lasers:  405 (diode), 457/488/514 (Argon Ion) 561 (diode); 640 (diode) (nm)
  • Dichroic mirrors: 405/488; 405/488/561; 405/488/561/640; 405-457/514; 457/514/561; BS20/80 (nm)
  • Emission filters: 450/50; 482/35; 525/50; 540/30; 595/50; 700/75 (nm)

For more information on

Dragon single point confocal: Semrock searchlight

Orion

Photo: LCI facility

This system can be used as

  • Spinning disk CREST v3
  • Ultra fast widefield with large field of view 
  • Super resolution (DeepSIM) ( approximately 100nm resolution in xy and approximately 300nm resolution in z (60x or100x oil objective); 1024x1024 FOV

Equipment specifications

  • Nikon
  • Inverted
  • Fully motorized including Z piezo 100 um
  • High content screening JOBS module
  • Camera from Photometrics,  Kinetix sCMOS, >95% QE, 6.5 um for best resolution, 2720x2720 pixels 

Filters, lasers and dichroic mirrors

  • Lasers: 405, 440, 475,  514, 545, 640, 748 (nm)
  • Dichroic mirrors: 441/30; 511/26; 592/37; 684/34; 817/66 (nm)
  • Excitation filters: 391/44; 477/12; 549/16; 639/17; 740/29
  • Emission filters: 440/40; 480/40; 525/50; 550/30; 595/50; 685/34; 750 LP (nm)

For more information on

Orion spinning disk/WF/DeepSIM Semrock searchlight

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: Kinetix sCMOS, >95% QE, 6.5 um for best resolution, 1193x1193 pixels
  • Spinning disk Yokagawa CSU-X1, 10,000 rpm
  • 1.2x built-in extra magnification lens in the spinning disk

Filters on the spinning disk confocal side

  • Lasers: 405, 488; 561; 640 (nm)
  • Dichroic mirrors: 431/44; 519/52; 599/55; 652LP (emission) (nm)
  • Emission filters: 447/60; 525/30 ; 607/36; 685/40; Dual band: 512/30; 630/100; Triple band: 525 600 690 (nm)

Filters on the widefield side

  • Excitation filters:  395/25, 440/20; 475/28; 510/25; 550/15; 575/25; 631/28 (nm)
  • Dichroic mirrors WF: 452/55; 529/52; 604/48; 675LP (emission) (nm)
  • Dichroic mirror CFP/YFP FRET: 458LP (emission) (nm)
  • Emission filters: 447/60; 483/32 ; 500LP; 525/30; 542/27; 572/15; 607/36; 624/40; 630/92; 685/40 (nm)

Filters on the TIRF side

  • Dual TIRF: Excitation filters:  488/561 (5 nm width); DM: 520/50; 565LP (emission) (nm); Emission filters: 522/45; 570LP (nm)
  • Quad TIRF: Excitation filters; 405; 488; 561; 647, 5 nm width; DM: 450/60; 525/60; 597/55; 607/55: 670LP (emission); Emission Filters: 451/29; 525/50; 607/18; 675LP (nm)

For more information on

Victor widefield: Semrock searchlight
Victor spinning disk: Semrock searchlight

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

Lasers

  • 405 (diode); 457/488/514 Argon; 561 (diode); 633 (diode)
    • 4 NDDs (2 trans and 2 epi) (specifications, in the table below)

For more information on

Zeus single and multipoint confocal: Semrock searchlight
Filters and DMs
Fluorophore Dichroic mirror Emission
Blue Mirror SP 485
Green LP 490 BP 500-550 used in combination with the blue filter cube
Green mirror mirror BP 500-550 used in combination with the red filter cube
Red LP 555 BP 565-610

Aurora

This system can be used as

  • Light sheet microscope

Equipment specifications

  • M Squared Lasers
  • Upright
  • Full size incubator for life cell imaging
  • Fully motorized including xy scanning size of a multiwell plate
  • Z fast beam scanning 900 um, additional stage scanning 1 cm
  • No shadowing effect thanks to the self-healing Airy beam
  • < 1um xy and z resolution with a field of view of 870x870 um
  • Tiling/stitching, multiple sample or regions
  • Dedicated deconvolution software running on GPU power
  • 2  Hamamatsu Orca Flash cameras 4 V.3 100 fps at 2048x2048 pixels. Pixel size 6.45 um. QE 82% 
  • Objective: Navitar multi-immersion: 15.3x – 17.9x; NA 0.37-0.43.
  • Refraction index: 1.33-1.56 without changing objectives
  • All immersion media can be used for imaging
  • Working distance 12mm

How to calculate the objective NA and magnification

  • NA=0.276xRI
  • Mag=11.494xRI
  • WD = constant for all RI

Lasers (mW)

  • 405nm(60 mW); 457nm(100 mW); 488nm(100 mW); 515nm(100 mW); 561nm(80 mW); 642nm(140 mW); 685nm(50 mW)

Filters

  • 445/58; 480/30; 525/50; 535/30; 590LP; 605/52; 665LP; 667/30; 700LP (nm)
    • DM: 510 LP (CFP-YFP) and 560 LP

A powerful server is available for fast transfer/storage/deconvolution/stitching/analysis.

For more information on

Aurora-light sheet microscopes
Aurora light sheet: Semrock searchlight

Minerva

  • 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
  • MMI VCXU23c colour camera (1920 x 1200 pixels, 5.86 μm x 5.86 μm)
  • MMI CellCut and CellEctor

Filters for widefield

  • Excitation filters: 365/10; 470/40; 544/25; 628/40 (nm)
  • Dichroic mirrors: 400LP; 495LP; 565LP; 660LP (nm)
  • Emission filters: 460/50; 515/30; 605/70; 692/40 (nm)

For more information on

Minerna widefield: Semrock searchlight link

Nikon AXR with NSPARC (single point confocal)

This system can be used as

  • Single point confocal
  • 25 mm FOV resonant scanner and 25 mm FOV galvano scanner

Equipment specifications

  • Nikon, Ti2
  • Inverted
  • Fully motorized including Z piezo 100 um
  • 2 low noise GaAsP and 2 Multi-alkali PMT detectors
  • Nikon Spatial Array Confocal (NSPARC) ultra-low noise detector
  • Extremely highspeed imaging (up to 720 fps at 2048 x 16 pixels)
  • Spectral detector in up to 66 emission channels for unmixing
  • High content screening JOBS module
  • Full size incubator for life cell imaging

Filters, laser and dichroic mirrors on the confocal side

  • Lasers: 405, 488, 561, 640 (nm)
  • Dichroic mirrors: 405/488/561/640; BS20/80 (nm)
  • Emission filters: fixed width: 429-474 and 663-728 and 2 tunable filters 
GI
Content reviewer:
Gabriela Imreh
10-11-2023