Microscopes at LCI

The Live Cell Imaging Core Facility offers high-end microscopes allowing researchers to perform the following microscopy modalities:

Below are the details for each microscope.

Tweety

This system can be used as 

  • Multipoint spinning disk confocal
  • Widefield 
  • Micropatterning (printing proteins on a dish) and micromanufacturing (making microwells)

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand with up to 32 mm FOV
  • Full size incubator for life cell imaging
  • Fast piezo z drive 100 um
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • Kinetix sCMOS camera over 95% QE, 6.5 um pixels for best resolution 
  • Alvéole Primo micropatterning and micromanufacturing device

Filters for both Widefield and Spinning disk side

Tweety widefield and spinning disk: Semrock searchlight

  • Lasers (nm): 
    • 405, 446, 477, 520, 546, 638, 749 
  • Excitation filters (nm): 
    • 391/44 
    • 477/12 
    • 549/16 
    • 639/17 
    • 740/32
  • Dichroic mirrors (nm): 
    • 441/30; 511/26; 593/37; 684/34; 817/66
    • 484/22; 561/30
    • 562LP
  • Emission filters (Single bands) (nm): 
    • 438/24 
    • 484/22 
    • 515/30
    • 562/40 
    • 593/40 
    • 685/40 
    • 561LP 
    • 809/81

Pegasus

This system can be used as

  • Single point scanning confocal with resonant scanner for fast imaging and spectral detector
  • Widefield
  • TIRF

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand
  • Full size incubator for life cell imaging
  • Fast piezo z drive 100 um
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • 2x GaAsP detectors (green and red)
  • 32-channel spectral detector
  • Andor EM-CCD camera, 95% QE, 60 fps at 512x512 pixels. Pixel size 16 um

Filters for the widefield side

Pegasus widefield: Semrock searchlight

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

Filters, lasers and dichroic mirrors for the confocal side

Pegasus single point confocal: Semrock searchlight

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

Filter cubes for TIRF

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

Dragon

This system can be used as

  • Single point scanning confocal with resonant scanner for fast imaging and spectral detector
  • Wide field with colour, monochrome and polarized light imaging

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand
  • Full size incubator for life cell imaging
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • 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

Filters, lasers and dichroic mirrors

Dragon single point confocal: Semrock searchlight

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

Orion

Photo: LCI facility

This system can be used as

  • Multipoint spinning disk confocal
  • Widefield
  • Super resolution (DeepSIM) ( approximately 100 nm resolution in xy and approximately 300nm resolution in z (60x or100x oil objective); 1024x1024 FOV

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand
  • Fast piezo z drive 100 um
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • Camera from Photometrics,  Kinetix sCMOS, over 95% QE, 6.5 um for best resolution

Filters, lasers and dichroic mirrors

Orion spinning disk/widefield/DeepSIM Semrock searchlight

  • Lasers (nm): 
    • 405, 446, 477,  518, 545, 637, 748 
  • Dichroic mirror (nm): 
    • 441/30; 511/26; 592/37; 684/34; 817/66 
    • 485/22; 561/30 (Dual)
  • Excitation filters (nm): 
    • 391/44
    • 477/12
    • 549/16 
    • 639/17 
    • 741/32
    • 449/20; 520/20 (Dual)
  • Emission filters (nm): 
    • 438/24 
    • 485/20 
    • 515/30
    • 562/40 
    • 593/40 
    • 685/40 
    • 809/81 

Victor

This system can be used as

  • Multipoint spinning disk confocal
  • Widefield
  • TIRF

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand
  • Full size incubator for life cell imaging
  • Fast piezo z drive 250 um
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • Widefield: 
    • Andor camera Zyla 4.2+. 82% QE, 100 fps at 2048x2048 pixels. Pixel size 6.45 um
    • External filter wheel and SpectraX LED for fast imaging
  • Spinning disk: Kinetix sCMOS, over 95% QE, 6.5 um pixel size
    • 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

Victor multipoint confocal: Semrock searchlight

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

Filters on the widefield side

Victor widefield: Semrock searchlight

  • Excitation filters (nm):  
    • 395/25, 440/20; 475/28; 510/25; 550/15; 575/25; 631/28 
  • Dichroic mirrors (nm):
    • 452/55; 529/52; 604/48; 675LP (emission)
  • Dichroic mirror (nm) (FRET): 
    • 458LP (emission) 
  • Emission filters (nm): 
    • 447/60
    • 483/32 
    • 500LP 
    • 525/30 
    • 542/27
    • 572/15
    • 607/3; 
    • 624/40 
    • 630/92 
    • 685/40 

Filters on the TIRF side

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

Aurora

This system can be used as

  • Light sheet microscope

Equipment specifications

  • 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, 82% QE, 100 fps at 2048x2048 pixels. Pixel size 6.45 um
  • 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)

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

Filters

Aurora light sheet: Semrock searchlight

  • Dichroic mirrors (nm): 
    • 510 LP and 560 LP
  • Emission filters (nm):
    • 445/50 
    • 480/30 
    • 525/50
    • 535/30 
    • 590LP 
    • 605/52 
    • 665LP 
    • 667/30
    • 700LP (nm)

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

For more information on

Aurora-light sheet microscopes

Minerva

  • This system can be used as

  • Widefield
  • Slide scanner
  • Laser microdissection

Equipment specifications

  • Nikon Ti2 inverted and fully motorized microscope stand
  • High content screening JOBS module
  • Smart Microscopy workflows using JOBS
  • MMI VCXU23c colour camera, 5.86 μm pixel size
  • MMI CellCut 

Filters for widefield

Minerva widefield: Semrock searchlight 

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

ECLIPSE Ji

ECLIPSE Ji
ECLIPSE Ji Photo: Gabriela Imreh

This system can be used as

  • Widefield

Equipment specifications

  • Benchtop microscope fully motorized 
  • 7.8 megapixel CMOS camera with 25 mm FOV
  • 0.38x built-in extra magnification lens
  • Preconfigured pipelines for plate assays such as: cell counting, cytotoxicity, intensity measurements, size and morphology, transfection efficiency. 

  • AI and Deep learning based acquisition settings and data analysis

    Filters for Widefield 

  • Excitation filters (nm):  
    • 378/52;  474/26; 554/23; 635/18
  • Dichroic mirrors (nm):
    • 432/36; 515/31; 595/31; 729/141 (emission)
  • Emission filters (nm): 
    • 432/36; 515/31; 595/31; 730/139
GI
Content reviewer:
Gabriela Imreh
27-06-2024