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:

Lasers: solid state 405, 488, 555, 639 Objectives: 5X(0.13), 10X(0.45), 20X(0.8), 40X(0.6), 40XW(1.2), 63XOil(1.4) Detector: PMT (2) Book Now
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Spinning Disc: Yokogawa X1 head with Borealis Lasers: solid state 405, 488, 552, 730 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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Spinning Disc: Diskovery/Nipkow spinning disk with dual-pinhole sizes  (50um & 100um) TIRF: Diskovery TIRF Unit Lasers: solid state 405, 488, 561,637 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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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:

Light Source: X-cite (Hg-halide), Colibri LED (455, 470, 505, 590) Cubes: Dapi, CFP, GFP, YFP, Cy3, Cy5, Cy7, mCherry 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, Zeiss AxioCam (colour) Book Now (Room 778)
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Light Source: Hg-halide Cubes: B, G, R 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:

Light Source: Xe-halide Cubes: Dapi, FITC, TRITC, Texas Red, Cy5 Objectives: 4X(0.2), 10X(0.45), 20X(0.5) Detector: CCD Camera – CoolSnap HQ Software: MetaXpress and AcuityXpress Book Now
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Slide Scanning:

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

Book Now

Zeiss AXIOSCAN

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.


Data Storage

The KRCBS Bioimaging Network Attached Storage (NAS) is a 60 TB capacity data storage unit that is on a shared network with all of the imaging facility computers, i.e. both acquisition and analysis workstation. The NAS was implemented to facilitate data transfer from acquisition computers (i.e. computers connected to microscopes) to the analysis computer workstations without the need for external hard drives. In addition, both RAW and processed data can be temporarily stored on the NAS for ongoing projects. For guidelines on how to access and use the Bioimaging NAS please refer to the following instructions.


Analysis Workstations

All analysis workstations are available for remote access using TeamViewer. Please see instructions here for remote access. Important reminder: you will still need to book the workstations using RFBMS for remote usage.

Analysis WorkStationRoomFree Software Licensed Software(s)
Ideal For
OSWindows UsernameTeam Viewer ID
FiJi
Imaris Scene
NIS Elements
Zen 3.2 Lite
Other
1552
xxxxView MSOT
MSOT photoacoustic data: Processing, visualization and analysis
Windows 10Workstation 1 User856083966
2552
xxxTscratchZen Black 2012, Zen Blue 2012
LSM700 data: FRAP and colocalization analysis
Windows 7Workstation 2 User307031234
3552
xxImaris 8.0.2 & 9.6.0, NIS Elements AR, Nikon Batch Deconvolution
Multidimesional data from all confocal systems: Deconvolution, 3D visualization, rendering and analysis, colocalization, particle tracking
Windows 10Workstation 3 User423614944
4552
xxxxQupath
LIFA
KUANT
Meta Imaging Series 7.8

Kuant
SD/TIRF data: Particle tracking (Meta Imaging Series 7.8)
Newton animal imaging analysis (Kuant)
Windows 10Workstation 4 User480470169
5778
xxHalo v2.3.2089.23
QuPath
Slide scanning data: Large 2D data set visualization and analysis
Windows 7 AxioScan User1095965369
6552
xMetaXpress
ImageXpress data: High content, low magnification, 2D image analysis
Windows 7 IXM User1657890043