Imaging Equipment

Online scheduling is available only to those individuals who have received training on equipment. Review the Bioimaging guideline on how to receive training. 

Acquisition Systems:

Confocal Systems:

ZEISS LSM 900 WITH AIRYSCAN
  • Lasers: 405nm, 488nm, 561nm, 640nm, 5mW
  • Objectives: 2.5x/0.085 WD=8.8mm, 10x/0.45 WD=2.0, 20x/0.8 WD=0.55mm, 25X/0.8 Multi Immersion, 63x/1.40 Oil Immersion
  • Detectors: 1 Array AiryScan detector, 2 GaAsP-PMT detectors
  • Filter cubes:
    • DAPI – Excitation 335-383; BP 395; Emission 420-470
    • GFP – Excitation 450-490; BP 495; Emission 500-550
    • DsRed – Excitation 538-562; BP 570; Emission 570-640
    • Cy5 – Excitation 625-655; BP 660; Emission 665-715
  • Book now (552)

QUORUM SPINNING DISC
  • Spinning Disc: Yokogawa X1 head with Borealis
  • Lasers: solid state 405, 488, 552, 730
  • Emission Filters: Dapi 447/60, GFP 525/50, YFP 545/40 , RFP/Cy3 593/40, TxRed 620/60, Cy7 810/90
  • Objectives: 10X (0.2), 20X (0.75), 40X (0.6 correction collar), 63X (1.4 oil), 100 (1.4 oil)
  • Detector:  Hamamatsu ImagEM X2 EM-CCD camera and ORCA 12-bit camera
  • Book Now (Room 778)
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QUORUM SPINNING DISC WITH TIRF
  • Spinning Disc: Diskovery/Nipkow spinning disk with dual-pinhole sizes  (50um & 100um)
  • TIRF: Diskovery TIRF Unit
  • Lasers: solid state 405, 488, 561,637
  • Emission filters: Dapi 450/50, GFP 525/50, RFP/Cy3 600/50, RED 610/75, Cy5 700/75
  • Objectives: 10X(0.25), 20X(0.40), 63XOil(1.47; CORR TIRF)
  • Detectors: Hamamatsu  ImagEM X2 EM-CCD camera, and ORCA-Flash 4.0 V2 PLUS sCMOS camera
  • Book Now (Room 778)
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NIKON A1R: RESONANT SCANNER AND MULTIPHOTON SYSTEM

Objectives:

  • Apochromat Multi-Photon LWD 25x Water Immersion Objective Lens, N.A. 1.1, W.D. 1.43-2.04mm, F.O.V. 22mm, DIC, Correction Collar 0-0.17mm
  • Apochromat TIRF 60X Oil Immersion Objective Lens, N.A. 1.49, W.D. 0.12mm, F.O.V 22mm, DIC
  • Plan Apochromat 10X Glycerol Immersion Objective Lens, N.A. 0.5, W.D. 5.5mm

Confocal Specifications:

  • Lasers: 405,488,561,640
  • Filters: 450/50, 525/50, 600/50, 650LP
  • Descan detector: 2 GaAsP and 2 High Sensitivity PMTs

Mutliphopton Specifications:

  • IR laser: Chameleon Vision II tuning range is 680-1080 nm, total output power at 680 nm is >650 mW and >3500 mW at 800 nm, pulse duration is 140 fs
  • Filters: 525/50, 605/70
  • Non descan detectors: 3 GaAsP and 1 High Sensitivity PMT

Book Now (Room 778)

Widefield Systems:

ZEISS WIDE FIELD WITH DECONVOLUTION
  • Light Source: Colibri 7 LED
  • Filter Cubes:
    • Dapi –  Excitation 333-383; BP 395; Emission 420-470
    • GFP –  Excitation 450-490; BP 495; Emission 500-550
    • DsRed –  Excitation 538-562; BP 570; Emission 570-640
    • mPlum – Excitation 574-599; BP 605; Emission 612-682
    • Cy5 –  Excitation 625-655; BP 660; Emission 665-715
  • Objectives: 5X(0.13), 10X(0.45), 20X(0.8), 40X(0.6), 40XW(1.2), 63XOil(1.4)
  • Detector: Axiocam 712 mono 4096 x 3008
  • Book Now (Room 552)

 

ZEISS AXIOOBSERVER
  • Light Source: X-cite (Hg-halide), Colibri LED (455, 470, 505, 590)
  • Emission Filter Cubes:
    • Dapi –  Excitation 350/50; Emission 460/50
    • CFP –  Excitation 436/20; Emission 480/40
    • GFP –  Excitation 470/40; Emission 525/50
    • YFP –  Excitation 500/20; Emission 535/30
    • Cy3 –  Excitation 545/25; Emission 605/70
    • mCherry –  Excitation 560/40; Emission 630/75
    • Cy5 –  Excitation 620/60; Emission 770/75
    • Cy7 –  Excitation 710/75; Emission 795/50
  • Objectives: 5X(0.13), 10X(0.3), 20X(0.4), 25X(0.8), 40X(0.6), 40XOil(1.4), 63XOil(1.4), 100XOil(1.4)
  • Detector: OrcaR2 CCD
  • Book Now (Room 778)
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OLYMPUS BX50
  • Light Source: Hg-halide
  • Cubes: Dapi, GFP, Cy3
  • Objectives: 4X (0.1),10X (0.25),20X (0.5),40X (0.75),60XOil (1.25),100XOil (1.3)
  • Detector: Olympus DP72 colour camera
  • Book Now
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High Content Imaging:

AGILENT/BIOTEK CYTATION 5
  • Sample formats – plates (6-1536 well), slides, dishes and flasks
  • Imaging Modes: Fluorescence, brightfield, high contrast brightfield, color brightfield, phase contrast
  • Excitation LED: 365, 465, 505, 555, 623, 740
  • Emission Filter Cubes:
    • Dapi –  Excitation 377/50; Emission 447/60
    • GFP –  Excitation 469/35; Emission 525/39
    • YFP –  Excitation 500/24; Emission 542/27
    • TRITC –  Excitation 556/20; Emission 600/37
    • Cy5 –  Excitation 628/40; Emission 685/40
    • Cy7 –  Excitation 710/40; Emission 809/81
  • Objectives (All Plan Fluorite):
    • 4X/0.13, WD: 17mm
    • 4/0.13 Phase, WD: 17mm
    • 10X/0.3 Phase, WD: 10mm
    • 20X/0.45 Phase, WD: 6.6-7.8nm
    • 40X/0.6 Phase, WD: 2.7-4.0 nm
  • Camera: Sony IMX264 CMOS, WFOV
  • Incubation: temperature to 65°C, CO2/O2
  • Accessories: BioSpa – allows for incubation and automated imaging for up to 8 plates
    • Incubation: temperature to 45°C, CO2/O2, humidity

ZEISS CELL DISCOVERER 7
  • Sample formats – plates (6-384 well), slides and dishes
  • Imaging Modes: Fluorescence, brightfield, phase gradient contrast, oblique contrast
  • Excitation LEDs:
    • Set 1: 385, 470, 567, 625
    • Set 2: 420, 520, 590
  • Emission Filters:
    • Quad Band Pass: 425/30, 514/30, 592/25, 709/100
    • Triple Band Pass: 467/24, 555/25, 687/145
  • Objectives (All Plan Apochromatic):
    • 5x/0.35 (2.5x/0.12; 5x/0.25; 10X/0.35), WD: 5.1mm
    • 20x/0.7 autocorr (10x/0.35; 20x/0.7, 40x/0.7), WD: 2.2mm
    • 20x/0.95 autocorr (10x/0.5; 20x/0.8; 40x/0.95), WD: 0.76mm
  • Camera: Axiocam 712 mono CMOS
  • Incubation: temperature to 60°C, CO2, humidity

Slide Scanning:

ZEISS AXIOSCAN.Z1

Slide scanning will be offered as a service, please see attached instructions and requisition form.

Objectives:

  • 2.5x/0.12 Fluar
  •  5x/0.25 Plan Fluar
  • 10x/0.45 Plan Apochromat
  • 20x/0.8 Plan Apochromat
  • 40x/0.95 Plan Apochromat

Brightfield illumination source:

  • white light LED

Brightfield Camera – Hitachi 3-Chip color camera, model HV-F202SCL:

  • pixel size of 4.4um, (0.22um pixel size at 20x)
  • Full 30 fps
  • 24, 30 or 36 bit depth

Fluorescent Illumination (COLIBRI LED):

  • Red (630nm) for excitation of Alexa 631, Cy5, TOTO-3 and similar dyes
  •  Yellow (590nm) for excitation of Alex 568, Alexa 594, mCherry, mPlum and similar dyes
  • Green (555nm) for excitation of Alexa 555, Cy3, TRITC, DsRed and similar dyes
  • Cyan (511nm) for excitation of Alexa 514, eYFP, Eosin, TOTO-1 and similar dyes
  • Blue (475nm) for excitation of Alexa 488, eGFP, Fluo4, FITC and similar dyes
  • Violet (430nm) for excitation of Alexa 430, eCFP, Lucifer Yellow and similar dyes
  • UV (385nm) for excitation of Alexa 405, DAPI, Hoechst 33258 and similar dyes

Fluorescent emission filters:

  • Blank for TL
  • Quad (90HE): 425/35, 512/35, 590/25, 660LP
  • Tri (91HE): 435/25, 555/25, 610LP
  • Dual: 520/40, 595/70
  • Single band green emission: 520/40
  • Single band red emission: 620/60

Fluorescent Camera- Hamamatsu ORCA-Flash 4.0 V3:

  • 16 bit
  • 6.5um pixel size
  • QE of 82%
  • maximum capture rate of 100f

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Zeiss AXIOSCAN
Analysis Software - HALO (Indica Labs)

5 HALO modules, two standard modules and three elective modules

  • Area Quantification Fluorescence – Analyze up to three fluorophores in a single image. Calculate percentage area and intensity of each fluorophore across analysis area and colocalization of mutltiple fluorophores
  • Area Quantification Brightfield – Separate up to three stains in a single image. Calculate percentage area of each stain, intensity of each stain, and colocalization of multiple stains
  • Tissue Classifier – Identify multiple tissues classes using a learn-by-example approach. Can be used in conjuction with all other modules (fluorescent and brightfield) to select specific tissue classes for further analysis.
  • Multiplex IHC – Quantify expression of up to 4 IHC markers in the nucleus, cytoplasm, or membrane. Ideally multiplex IHC applications.
  • High-Plex FL – Cell based analysis for up to 8 markers (plus DAPI). Ideally suited for high-plex FL applications and multi-spec imaging systems.