Microscopy & Histology Core Facility

VisiScope Spinning Disk Confocal Microscope – live (Room -1.261) 

The Spinning Disc Confocal (5 Elements, Visitron) microscope has an ablation laser on a Nikon stand with a CSU-W1 spinning disk (25 and 50 µm pinholes) from Yokogawa. It is equipped with two sCMOS cameras for simultaneous live cell imaging in two channels and seven laser lines. The microscope is optimised for live cell imaging with TIRF, FRAP and DNA ablation options (355 nm pulsed laser). 

•  Available objectives: 10x/0.45 air, 10x/0.5 gly/water/oil, 20x/0.75 air, 40x/0.95 air, 20x/0.95 water, 25x/1.05 silicone, 40x/1.25 water, 60x/1.2 water, 100x/1.49 oil 

BC43Spinning Disk Confocal Microscope – live (Room 00.413) 

The BC43 (Andor) is a prime solution for 3D imaging thick samples. A high-sensitivity, high-speed imaging camera makes it perfect for live-cell confocal imaging. Widefield and transmitted light modes are available as well. A motorised XY stage allows multiple-position acquisition, large organism montages and multipoint scanning, resulting in outstanding productivity from a single imaging experiment. 

• Epi and confocal fluorescence mode 
• Four excitation lines (405, 488, 561, 640nm) 
• Available objectives: 2x air, 10x/0.45 air, 20x/0.75 air, 40x/0.75 air, 60x (1.4) oil

Stellaris 8 FALCON Confocal Laser Scanning Microscope with Fluorescence Lifetime Option (Room 00.421) 

The Stellaris 8 FALCON (Leica) is an inverse confocal fluorescence scanning microscope with the capability to perform Fluorescence Lifetime Imaging Microscopy (FLIM). It is optimal for imaging living and fixed samples with multicolor fluorescence.

• FLIM can be used for several applications, such as improving image quality by removing autofluorescence, multiplexing fluorophores with overlapping spectra, detecting cellular metabolites based on their fluorescence lifetime, and measuring molecular interactions with FRET sensors
• Deconvolution module (Lightning) 
• FRAP module 
• Fast scanning with a resonance scanner 
• Equipped with a pulsed white light laser (excitation range 440-790 nm, 80MHz pulse frequency) and diode lasers (405 nm, 488 nm, 561 nm) 
• The fluorescence detection range can be continuously adjusted between 400 - 850 nm and emission is detected by optimized photomultipliers (HyD-S, HyD-X, HyD-R)
• Mosaic scanning of large objects is possible due to the motorised xy-stage 
• Available objectives: 10x/0.4 air, 20x/0.75 air, 40x/1.3 oil, 63x/1.4 oil, 100x/1.4 oil, 63x/1.2 water 

SP5 Confocal Laser Scanning Microscope (Room 00.461) 

The TCS SP5 (Leica) is an inverse confocal fluorescence microscope optimised for imaging fixed samples with multicolour fluorescence. Four lasers are available for excitation (405, argon (458, 476, 488, 495, 514), 561 and 633 nm). The fluorescence detection range can be continuously adjusted between 400 - 800 nm and emission is detected by 4 PMTs. Mosaic scanning of large objects is possible due to the motorised xy-table. 

•  Available objectives: 10x/0.3 dry, 20x/0.7 dry, 40x/1.3 oil, 63x/1.2 water, 63x/1.4 oil

SP5 STED CW Super-Resolution Microscope (Room -1.261) 

The TCS SP5 STED CW (Leica) confocal microscope is dedicated to super-resolution microscopy based on STED (STimulated Emission Depletion) and live cell imaging. Super-resolution in the xy-direction (ca. 70 nm) is achieved for fluorophores with emission spectra overlapping with the 592 nm depletion laser. Additionally, the setup is equipped with an FCS module (Picoquant) and APD sensors to measure diffusion rates. Three lasers with the following wavelengths are available for excitation: argon (458, 476, 488, 495, 514), 561 and 633 nm. The detection range can be continuously adjusted between 400 - 800 nm and emission is detected by 3 PMTs and 2 HyDs. 

• Available objectives: 10x/0.3 dry, 20x/0.7 water/oil, 40x/1.1 water, 63x/1.2 water, 63x/1.4 oil, 100x/1.4 oil 

CX-F 3D Cell Explorer - live (Room -1.291) 

The 3D Cell Explorer fluo (Nanolive) is a holo-tomographic widefield microscope with fluorescence options for gentle live cell imaging. The contrast is based on refractive index measurements in living samples and provides "label-free" discrimination of several organelles (mitochondria, lipid vesicles, nuclei, nucleoli, lysosomes, etc.). 

Whole 3D volumes as large as 80 µm x 80 µm x 30 µm can be imaged within 2 seconds. The set-up is equipped with a 60x air objective and an incubation chamber (temperature, CO₂ settings available) for live cell imaging. Fluorescence imaging options (widefield) for green, red and far-red fluorophores are available.

IncuCyte SX5 Widefield Fluorescence Screening Microscope - live (Room 00.281) 

The IncuCyte SX5 (Sartorius) is a box-type widefield screening microscope for live cell imaging with several predefined assays and analysis pipelines. The setup is equipped with 4x, 10x and 20x air objectives and located inside an incubator chamber (temperature, CO₂ settings available) ideally for long-term live cell imaging. There are six slots for multiwell plates (6, 12, 24, 48 or 96 well) and/or four slots for chambered slides. 

Two fluorescence detection modules are available: 

• PH/G/R: Phase contrast/green (441-481 nm, 503-544 nm)/red (567-607 nm, 622-704 nm) 
• PH/G/O/NIR: Phase contrast/green (453-485 nm, 494-533 nm)/orange (546-568 nm, 576-639 nm)/NIR (648-674 nm, 685-756 nm) 

Opera Phenix Spinning Disk Confocal Microscope – live (Room 00.531) 

The Opera Phenix (Revvity) is a high-content screening (HCS) microscope system, built to handle multi-well plates as well as slides. The system can be operated in widefield mode and spinning-disc confocal mode. Fixed and living cells/organisms can be recorded. The system is equipped with two sCMOS cameras for fast recording, with four laser lines (405, 488, 561 and 640 nm). 

• Available objectives: 1.25x/0.03 air, 10x/0.3 air, 20x/1.0 water, 40x/1.1 water, 63x/1.15 water 

NanoImager Super-Resolution Microscope (Room 00.413) 

The NanoImager is a compact benchtop widefield microscope dedicated to single-molecule localization microscopy (SMLM). Resolution in the range of 20 nm in xy (STORM, DNA-PAINT, PALM) and 100 nm in Z (+/- 600 nm) can be obtained using this method. SMLM in multiple colors (four lasers) and two simultaneous channels (dichroic mirror split at 640 nm) is possible. 

• Excitation laser power at sample: 405 nm (15 mW), 488 nm (250 mW), 561 nm (250 mW), 640 nm (250 mW) 
• Excitation laser power at source: 405 nm (1 W), 488 nm (1 W), 561 nm (750 mW), 640 nm (1 W) 
• Available objective: 100x/1.45 oil 
• Motorised xyz stage
• TIRF/HiLo mode

GSDIM Super-Resolution Microscope (Room -1.261) 

The GSD 3D (Leica) widefield microscope is dedicated to super-resolution microscopy based on fluorophore blinking achieved by GSD (Ground State Depletion) and the single-molecule localisation algorithm (also called dSTORM or PALM). Structures in the range of 40 nm (xy) and 50 nm (z) can be resolved. The microscope is further equipped with a TIRF (total internal reflection) module, which is ideal for imaging samples located close to the cover glass/water interface, such as membrane receptors. The objective for super-resolution and TIRF is 160x/1.47 oil. 

SP5 STED CW Super-Resolution Microscope (Room -1.261) 

The TCS SP5 STED CW (Leica) confocal microscope is dedicated to super-resolution microscopy based on STED (STimulated Emission Depletion) and live cell imaging. Super-resolution in the xy-direction (ca. 70 nm) is achieved for fluorophores with emission spectra overlapping with the 592 nm depletion laser. Additionally, the setup is equipped with an FCS module (Picoquant) and APD sensors to measure diffusion rates. Three lasers with the following wavelengths are available for excitation: argon (458, 476, 488, 495, 514), 561 and 633 nm. The detection range can be continuously adjusted between 400-800 nm and emission is detected by 3 PMTs and 2 HyDs. 

• Available objectives: 10x/0.3 dry, 20x/0.7 water/oil, 40x/1.1 water, 63x/1.2 water, 63x/1.4 oil, 100x/1.4 oil

FRAP/TIRF/Ablation: VisiScope Spinning Disk Confocal Microscope – live (Room -1.261) 

FRAP/FLIM: Stellaris 8 FALCON Confocal Laser Scanning Microscope with Fluorescence Lifetime Option (Room 00.421) 

FRAP/FCS: SP5 STED CW Super-Resolution Microscope (Room -1.261) 

CX-F 3D Cell Explorer - live (Room -1.291) 

The 3D Cell Explorer fluo (Nanolive) is a holo-tomographic widefield microscope with fluorescence options for gentle live cell imaging. The contrast is based on refractive index measurements in living samples and provides "label-free" discrimination of several organelles (mitochondria, lipid vesicles, nuclei, nucleoli, lysosomes, etc.). 

Whole 3D volumes as large as 80 µm x 80 µm x 30 µm can be imaged within 2 seconds. The set-up is equipped with a 60x air objective and an incubation chamber (temperature, CO₂ settings available) for live cell imaging. Fluorescence imaging options (widefield) for green, red and far-red fluorophores are available. 

Lumicks C-Trap (Room –1.281) 

The C-Trap (Lumics) combines high-resolution optical tweezers with advanced microfluidics and multicolor confocal imaging, allowing manipulation and visualisation of single molecules and the measurement of piconewton forces. The system is designed for automated control over the flow of up to five solutions, with four optical traps and confocal imaging with three laser excitation lines (488, 561 and 638 nm). 

The Huygens Remote Manager provides access to web-based deconvolution by the Huygens Software (SVI). Here you can upload your images to the university server and start the deconvolution. You will receive an email notification when the deconvolution has finished. 

Five image processing stations (Room 00.457)

Both commercial (Imaris, vision 4D, VisiView, LAS-X, Zen blue Intellesis, Huygens and Huygens Remote Manager, Harmony, Columbus, GraphPad Prism, MatLab) and open source (e.g. Fiji (ImageJ), QuPath, Icy) software tools are available, on-site and remotely. 

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