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
• 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 (Single bands) (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

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 and an  built-in 1.8x lens 

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 (Single bands) (nm):
  • 450/50
  • 482/35
  • 525/50
  • 540/30
  • 595/50
  • 700/75 

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 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)
• Emission filters (nm):
  • 438/24
  • 484/22
  • 515/30
  • 562/40
  • 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 

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
• Available objectives: 4x air (NA 0.20), 10x air (0.5), 20x air (0.8)

Available pre-configured assays pipelines

• Intensity measurement
• Cell counting (end point): DAPI or Hoescht staining
• Cytotoxicity: measures live and dead cells and the ratio (Hoescht and ethidium homodimer 1))
• Cell size and morphology analysis - protocol
• AI-powered optimization of transfection efficiency by nucleus and cellular region detection from brightfield images: protocol
• Apoptosis assay: protocol
• Quantitative analysis of autophagy activity assay: protocol
• Analysis of DNA Damage (gamma-H2AX): protocol
• Analysis of nuclear translocation: protocol 
• Calculating the percentage of cells with micronuclei: protocol
• Analysis of endocytosis: protocol
• Analysis of phagocytosis: protocol
• Mitochondrial toxicity analysis: protocol

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