Cellular imaging

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 

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 

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 

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 

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 

Inverted laser-scanning confocal microscope – 4 lasers. 

Description to come.

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. 

Software available: 

• Imaris Full Spectrum and deconvolution module, latest version 
• AutoQuant V3 for deconvolution 
• FIJI/ImageJ 
• Zen Blue Lite 
• LasX Lite