Cellular imaging
Equipment
Our core facility offers a range of optical microscopy equipment to meet your cellular imaging needs. Our staff is committed to providing the best service possible by guiding you in selecting the most appropriate microscope for your applications.
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Semi-automated, widefield upright microscope for brightfield slide image acquisition.
Transmitted light (TL) modes available:
- Brightfield (BF)
- Phase contrast (Ph)
- Darkfield (DF)
- Condensor with Ph and DF filters
Objectives:
- Manual nosepiece
- N Plan 2X/0.06
- Fluotar 10X/0.3 Ph1
- Fluotar 20X/0.5 Ph2
- Fluotar 40X/0.75 Ph2
- Fluotar 100X/1.3Oil Ph3 (upon request)
Camera:
- Leica DFC300 FX colour camera (RGB)
- 1392 x 1040 pixels in Full Frame HQ mode; possible 2088 x 1560 pixels with 1.5x scaling
- White balance
- 3 x 8 bits
XY Stage:
- Manual
- Slide holder
Software:
- LAS, Version 3.6
- Generation of .tiff images with metadata (other formats available: Jpeg, Jpeg2000, PNG, Bmp, psd)
Possible applications:
- Taking brightfield photos (BF, Ph and DF)
- Adapted for fixed, stained samples
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Inverted spectral confocal laser-scanning microscope – 3 lasers (Leica Microsystems, TCS SP5)
Possible applications:
- Spectral confocal (300 nm-800 nm)
- ZStacks
- 2D and 3D projections
- Field rotator
- Lambda stacks (3 nm precision and minimum 5 nm bandpass)
- Spectral deconvolution
- FRET
- FRAP
Objectives:
- PL Fluotar 10X/0.3
- PL Fluotar 20X/0.5
- PL Apo 40X/1.25 Oil
- PL Apo 63X/1.4 Oil
- PL Apo 100X/1.4 Oil
Lasers:
- Argon laser (458, 476, 488, 514 nm) with acousto-optical tunable filters (AOTF)
- DPSS 561 with AOTF
- HeNe 633 with AOTF
Detectors:
- 3 PMTs (spectral, 300-800 nm, positions 1,2 and 4))
- 1 HyD (spectral, 300-800 nm, position 3)
- TLD for capturing brightfield images (BF, DIC, POL)
Stage:
- Manual XY
Software:
- LAS-AF (Version 2.7.1)
- FRET and FRAP wizard modules
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Inverted spectral confocal laser-scanning microscope – 4 lasers and motorized stage (Leica Microsystems, TCS SP5).
Possible applications:
- Spectral confocal (300 nm-800 nm)
- ZStacks
- 2D and 3D projections
- Field rotator
- Lambda stacks (3 nm precision and minimum 5 nm bandpass)
- Spectral deconvolution
- FRET
- FRAP
- Mosaic and multiposition
Objectives:
- PL Fluotar 10X/0.3
- PL Apo 20X/0.7 IMM Corr
- PL Apo 40X/1.25 Oil
- PL Apo 63X/1.4 Oil
- PL Apo 100X/1.4 Oil
Lasers:
- 405 nm laser diode with filter wheel (3/6/12,5/25/50/80/100%)
- Argon laser (458, 476, 488, 514 nm) with acousto-optical tunable filters (AOTF)
- DPSS 561 with AOTF
- HeNe 633 with AOTF
Detectors:
- 3 PMTs (spectral, 300-800 nm, positions 1,2 and 4)
- 1 HyD (spectral, 300-800 nm, position 3)
- TLD for capturing brightfield images (BF, DIC, POL)
Stage:
- Motorized with Galvo-Z
Software:
- LAS-AF (Version 2.7.1)
- FRET and FRAP wizard modules
- Stitching
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Upright confocal multi-photon laser-scanning microscope – 4 visible lasers and one Ti:Sapphire Mai-Tai (Spectra-Physics) laser. Incubation chamber for temperature control, motorized stage and DeepLens water immersion objectives.
This microscope is usually dedicated to small animal intravital microscopy.
Possible applications:
- 2-photon laser (690-1040 nm)
- Incubation chamber (temperature-controlled)
- Intravital core (mouse-rat)
- 3NDD (Non-Descanned Detectors/RXD)
- Confocal
- ZStacks
- 2D and 3D projections
- Scan Field rotator
- Mosaic and multiposition
Objectives:
- UPlan SApo 4X/0.16
- UPlan SApo 10X/0.4
- UPlan SApo 20X/0.75
- UPlan SApo 40X/0.95
- UPlan SApo 60X/1.35 Oil
- DeepLens XLUM Plan FL N 20X/1.00 W (for intravital)
- DeepLens LUM FL N 60X/1.10 W
Lasers:
- 405 nm laser diode with acousto-optical tunable filters (AOTF)
- Argon laser (458, 488, 515 nm) with AOTF
- 543 nm laser with avec AOTF
- 635 nm laser with AOTF
- Mai-Tai (Spectra-Physics) 690-1040 nm tunable multi-photon
Detectors:
- 4 PMTs with filters:
- Ch. 1: SMD 490/510/560 ; BA 430-470 / 480-495 / 505-525
- Ch. 2: SMD 560 ; BA 505-525 / 505-605 / 535-565
- Ch. 3: SMD 640 ; BA 560-620 / 590-660
- Ch. 4: Mirror ; BA 655-755
- 3 RXD (NDD) with filters for 2-photon microscopy
- TLD for capturing brightfield images
Stages:
- Motorized stage
- Adjustable height
Software:
- Fluoview FW10-ASW (version 3.01.03.03)
- Stitching
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Inverted spinning disk (SD) confocal microscope with TIRFM module, incubation chamber, Definite Focus 2 module and Piezo Z- Focus Drive.
Real-time microscopy applications have priority use of this microscope.
This microscope may sporadically be used for risk group 3 pathogens (HIV/HCV).
Possible applications:
- Conventional fluorescence
- Spinning disk confocal acquisitions
- TIRFM
- Real-time videomicroscopy experiments
- Zstacks
- 2D/3D projections
- Fast acquisitions through hardware synchronization
- DIC
Objectives:
- EC Plan NeoFluar 10X/0.3 DICI
- LD PlanNeofluar 20X/0.4 DICII
- Plan Apochromat 20X/0.8 DICII
- Fluar 40X/1.3 Oil DICIII
- α Plan Apochromat 100X/1.46 Oil DICIII TIRF
Light source for conventional fluorescence:
- X-Cite 120Q LED (Excelitas Technologies)
Filters for conventional fluorescence:
- Filter Set 49 (G365 – FT395 – BP445/50) (DAPI, Hoechst, Alexa405)
- Filter Set 38 HE (BP470/40 – FT495 – BP525/50) (GFP, Alexa488, FITC)
- Filter Set 43 HE (BP550/25 – FT570 – BP605/70) (DsRed, Cy3, RFP, mCherry, Alexa555, Alexa568)
- Filter Set 64 HE (BP587/25 – FT605 – BP647/70) (Alexa594, TexRed)
- Filter Set 50 (BP640/30 – FT660 – BP690/50) (AF647, Cy5)
- Filter Set ET-Cy7 (BP710/75 – FT760 – BP810/90) (Alexa700+)
- Filter Set 76 HE (TBP406+489+561 – TFT427+503+578 – TBP459+525+608)
- Filter Set 77 (TBP482+563+642 – TFT503+582+658 – TBP524+604+688)
Lasers for spinning disk confocal and TIRFM modes:
- 405 nm (50 mW) SSL with acousto-optic tunable filters (AOTF)
- 488 nm (20 mW) SSL with AOTF
- 561 nm (40 mW) SSL with AOTF
- 639 nm SSL with AOTF
Spinning disk module:
- Yokogawa CSU-X1
Detection filters:
- DBP 480/22 + LP530
- DBP 527/54 + 645/60
- BP 690/50
- FF01 446/523/600/677
- DBP 460/30 + 590/30
- DBP 530/30 + LP650
Cameras:
- CCD AxioCam MRm for fluorescence and TIRF
- 14-bit monochrome camera
- 1388 X 1040 pixels
- 6.45µm pixel size
- EMCCD Evolve 512 (Photometrics) for the spinning disk confocal mode
- 16-bit monochrome camera
- 512 x 512 pixels
- 16µm pixel size
- High sensitivity
Stage:
- Motorized
- Equipped with ASI
- MCL nano-drive piezo (Focus drive)
Incubation chamber:
- Ibidi: Temperature and CO2 /adaptors for culture dishes (6 cm) or slides (multi-well or channels)
- Tokai Hit: heating ring for objectives
Software:
- ZEN Blue (Version 2012)
- Tiling and Stitching modules
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Inverted laser-scanning confocal microscope – 4 lasers.
Description to come.
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Upright widefield microscope for observing slides in fluorescence (three colours), as well as in brightfield. No camera attached.
Objectives:
- 10X/0.4
- 20X/0.75
- 40X/0.95
- 60X/1.3 Water
Filters:
- DAPI, Hoesch, Alexa405
- GFP, FITC, Alexa488
- TxRed, mCherry, Alexa594.
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Software available:
- Imaris Full Spectrum and deconvolution module, latest version
- AutoQuant V3 for deconvolution
- FIJI/ImageJ
- Zen Blue Lite
- LasX Lite