Microscopes at LCI

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
• Crest Optics X-Light V3 spinning disk (50 um pinholes with 250 um interval)
• 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
• Dichroic mirrors (nm):
  • 441/30; 511/26; 593/37; 684/34; 817/66
  • 484/22; 561/30
  • 562LP
• Emission filters (nm):
  • 438/24
  • 484/22; 561/30
  • 515/30
  • 562/40
  • 593/40
  • 685/40
  • 561LP
  • 809/81

This system can be used as

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

Equipment specifications

• Nikon Ti2-E inverted and fully motorized microscope stand
• Stage top incubator
• Resonant scanner
• Fast piezo z drive (100 um range)
• High content screening JOBS module
• Smart Microscopy workflows using JOBS
• 2x tunable GaAsP detectors (400-750 nm)
• 1 Multi Alkali PMT detector (blue)
• 1 Extended Red GaAsP detector (Far Red, NiR)
• Spectral detector
• Digital Sight 10 CMOS colour camera from Nikon, 25 mm FOV, 5.97 pixel size and a built-in 1.8x lens

Lasers and dichroic mirrors to Ax confocal

• Lasers (nm):
  • 405; 488; 561; 638, 730
• First dichroic mirror:
  • 405/488/561/640/730
  • 405/488/561/640
  • BS20/80 (400-750)

Filter cubes to Ax confocal detectors

• Multi Alkali PMT detector:
  • Cube1: DM: 495LP; Em: 452/45
  • Cube2: DM: 550/300, 750LP; Em: 445/50, 675/30
• Extended Red GaAsP detector:
  • Cube1: DM: 640SP; Em: 700/75
  • Cube2: DM: 724SP; Em: 740LP

Pegasus: filters and lasers setup on Semrock Searchlight

This system can be used as

• Single point scanning confocal with resonant scanner for fast imaging and spectral detector for channel unmixing
• NSPARC super-resolution (100 nm)

Equipment specifications

• Nikon Ti2-E inverted and fully motorized microscope stand
• Stage top incubator
• Resonant scanner
• Fast piezo z drive (100 um range)
• High content screening JOBS module
• Smart Microscopy workflows using JOBS
• 2x tunable GaAsP detectors (400-750 nm)
• 1 Multi Alkali PMT detector (blue)
• 1 Extended Red GaAsP detector (Far Red, NiR)
• Spectral detectors
• NSPARC detector (Single Photon Avalanche Diode)

Filters, lasers and dichroic mirrors AX confocal

• Lasers (nm):
  • 405; 445; 488; 514; 561; 638, 730
• First dichroic mirror:
  • 405/488/561/640/730
  • 405/488/561/640
  • 405/445/515/561/640
  • BS20/80 (400-750)
  • 445/515/561

Dragon AX: filters and lasers setup on the Semrock Searchlight.

Filter cubes specific to Ax confocal detectors

• Multi Alkali PMT detector:
  • Cube1: DM: 495LP; Em: 452/45
  • Cube2: DM: 550/300, 750LP; Em: 445/50, 675/30
  • Cube3: DM: 515/LP; Em: 482/35
• Extended Red GaAsP detector:
  • Cube1: DM: 640SP; Em: 700/75
  • Cube2: DM: 724SP; Em: 740LP

Emission filters specific to NSPARC detector

• Quad: 446/33; 524/42; 600/36; 677/28
• Single bands: 452/45; 482/35; 525/50; 540/30; 593/46; 700/75; 750LP

Dragon AX-NSPARC: filters and lasers setup on the Semrock Searchlight.

This system can be used as

• Multipoint spinning disk confocal
• Widefield
• Super resolution - DeepSIM

Equipment specifications

• Nikon Ti2 inverted and fully motorized microscope stand
• Crest Optics X-Light V3 spinning disk (50 um pinholes with 250 um interval)
• Super resolution (DeepSIM) ( approximately 100 nm resolution in xy and approximately 300nm resolution in z (60x or100x oil objective); 1024x1024 FOV
• Fast piezo z drive 600 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, 445, 488, 555, 638, 735
• Dichroic mirror (nm):
  • 446/60; 522/43; 595/49; 684/55; 812/113
  • 484/22; 561/30 (Dual)
• Emission filters (nm):
  • Penta-band: 446/53; 521/34; 593/40; 685/49; 819/91
  • Single-band:
    • 438/24
    • 485/20
    • 515/30
    • 593/40
    • 685/40
    • 809/81

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 (single bands) (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.

• This system can be used as

• Widefield
• Slide scanner
• Laser microdissection
  • Laser microdissection of tissue samples
  • Selective Isolation of living Cells for Omics Analysis
  • Isolation of rare or single cells for further cultivation

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