Andor Revolutions XD

Contact: Tomasz Wegierski

Access: training required

General Description

Spinning-disc (Yokogawa CSU-X1) and TIRF (Zeiss, manual illuminator) imaging system on inverted stand. Lasers: 405, 488, 561, and 640 nm. Three front-illuminated EM-CCD Andor cameras. Lenses: 20×/0.5 (dry), 20×/0.75 (dry), 40×/1.3 (oil), 63×/1.4 (oil), 100×/1.46 (oil). Incubation cage and heating insert with CO2 delivery for live imaging. Hardware autofocus. Motorized XY stage and Piezo Z insert. Acquisition software: IQ3.

Applications

• Fast confocal volumetric imaging of fluorescently labeled live specimens (spinning-disc, Z piezo)
• Imaging of endocytosis (TIRF with 100×/1.46 lens)
• FRAP and photoactivation (FRAPPA module)
• Imaging at enhanced resolution (offline SRRF processing)

Sample preparation

Samples should be on glass coverslips (e.g. slides, 35mm petri dishes with glass bottom, 40mm Warner Instruments chambers). TIRF requires samples in aqueous media on high-quality 0.17 µm (“1.5H”) coverslip dishes. No special requirements for fluorescent dyes.

Fujitsu “Celsius” R940 Workstation

Contact: Tomasz Wegierski

Access: local account required

General Description

Workstation for image processing and analysis. 2x Intel Xeon CPUs, 128 GB RAM, 27” LED/IPS monitor. Storage on RAID0 (fast access) and RAID1 (archive) disk arrays. Software: Imaris 8.3. Possibility to work via Remote Desktop Protocol.

Applications

• 3D/4D image analysis and rendering

File preparation

czi, lsm, and tiff files are converted to ims files for the analysis in Imaris

Zeiss Lightsheet Z.1

Contact: Tomasz Wegierski

Access: training required

General Description

Lightsheet-type microscope for imaging of fluorescently labeled large samples (e.g. zebrafish larvae) with optical sectioning. Lasers: 405, 458, 488, 514, 561, and 640 nm. Two sCMOS cameras. Detection lenses: 5× (dry), 20×/1.0, 40×/1.0, and 63×/1.0 (all water immersion).  Samples can be moved in 4-axes. Dual-side excitation and pivot scan for better acquisition of obscured structures. Continuous Z drive for fast volumetric imaging. Acquisition software: ZEN 2014 for Lightsheet. Two 30-TB storage PCs for data acquisition.

Applications

• Fast volumetric imaging of fluorescently labeled (live) zebrafish
• Simultaneous imaging in two spectral channels
• Multiview imaging, registration, and fusion
• Possibility to image in transmitted light-mode

Sample preparation

Samples should be embedded in capillaries in 1-2% low-melting point agarose. Live zebrafish should be anaesthetized. Multiview imaging may require the addition of fiducial markers (fluorescent beads).

Nikon Eclipse 80i

Contact: Tomasz Wegierski

Access: training required

General Description

Wide-field fluorescence microscope on upright stand for monochromatic imaging of fluorescence and transmitted-light color imaging. Illumination with 100 W mercury lamp or halogen lamp. Four spectral channels: DAPI, FITC, mCherry/Texas Red, and Cy5. Lenses: 2×, 4×, 10×, 20×, 40× (all dry), 40× oil, 60× oil and 100× oil. Phase-contrast on some lenses. Monochromatic CCD camera with optional RGB filter wheel. Motorized XYZ axes, manual objective revolver and filter wheel. Acquisition software: Image-Pro Plus 7.0.

Applications

• Wide-field observation and imaging of fluorescent samples
• Mosaic imaging of histological sections in transmitted light

Sample preparation

No special requirements for sample preparation.

Olympus CellR/ScanR

Contact: Tomasz Wegierski

Access: training required

General Description

Wide-field fluorescence microscope on inverted stand for real-time acquisition of live and fixed cells. Illumination with 150 W xenon lamp or 100 W halogen lamp. Fast excitation filter-wheel and shutter. Front-illuminated EM-CCD Hamamatsu camera. Lenses: 10×/0.4, 20×/0.75, 40×/0.6, 40×/0.95 (all dry) and 60×/1.35 (oil). Nomarski contrast. Incubation cage and heating insert with CO2 delivery for live imaging. Hardware and software autofocus. Motorized XYZ axes. Acquisition software: CellR for multidimensional imaging and ScanR for unbiased, high-throughput imaging.

Applications

• Fast, multidimensional, fully motorized acquisition of fluorescence samples
• Ca2+ imaging with Fura-2
• Semi-high throughput imaging of cells in multiwell plates

Sample preparation

Samples should be on glass coverslips (e.g. slides, 35mm petri dishes with glass bottom, 40mm Warner Instruments chambers) or thin imaging-quality plastic except when using 10×/0.4 and 40×/0.6 which can image through thicker glass/plastic dishes (thickness must be less than 2 mm). No special requirements for fluorescent dyes.

Olympus IX70

Contact: Tomasz Wegierski

Access: training recommended but not required

General Description

Wide-field manual fluorescence microscope on inverted stand for viewing and imaging of fluorescently labeled samples. Illumination with 100 W mercury lamp or halogen lamp. Four filter cubes: blue, green, red and “white-light” cube to be used with CFP/YFP/dsRed excitation filters or for transmitted light. Lenses: 10×, 20×, 40× (all dry), and 100× oil. Phase-contrast on 10× – 40× lenses. Monochromatic CCD camera. Acquisition software: AnalySIS.

Applications

• observation and imaging of cells on slides and cell culture dishes, e.g. after transfection

Sample preparation

No special requirements for sample preparation. For 10× and 20× lenses, the cells can be in standard cell-culture dishes.

PerkinElmer Opera Phenix

Contact: Tomasz Wegierski

Access: training required for internal users,

full imaging service for external users

General Description

The Opera Phenix from PerkinElmer is an automated confocal spinning disk microscope. It is equipped with a robotic arm for automatic plate handling, two cameras and four laser lines (405, 488 561, 640 nm). The microscope has four air objectives (1.25×, 5×, 10× and 20×) and three water objectives with automatic immersion system (20×, 40×, and 63×). The system features “Harmony” software for unbiased high-content image analysis including machine learning algorithms for image segmentation and object classification.

Applications

• Confocal imaging of fluorescently labeled samples in multi-well plates
• Brightfield and digital contrast imaging
• Live imaging
• Co-localization

Sample preparation

Samples must be in multi-well plates suitable for microscopy. No special requirements for fluorescent dyes.

Zeiss LSM 710 NLO

Contact: Tomasz Wegierski

Access: training required

General Description

Confocal and 2-photon microscope on inverted stand Zeiss A×io Observer Z1. Lasers: 405 nm, Argon laser (458, 488, 514 nm), HeNe lasers (543, 594, and 633 nm), and MaiTai eHP DeepSee 2-photon laser (690-1040 nm). Detectors: two standard PMTs, one 32-channel spectral detector, and one T-PMT. Lenses: 10×/0.3 (dry), 20×/0.5 (dry), 40×/1.3 (oil), 40×/1.1 (water), 63×/1.4 (oil), 100×/1.4 (oil). DIC with lenses: 40×/1.1 and 40×/1.3. Incubation cage and heating insert with CO2 delivery for live imaging. Software and hardware autofocus. Motorized XY stage. Acquisition software: ZEN 2009. More advanced experiments can be designed with Multi-Time Series macro.

Applications

• Confocal imaging of fluorescently labeled specimens
• 2-photon imaging (best suited is 40×/1.1 UV-VIS-IR lens, W.D. 0.62 mm)
• Live imaging
• FRAP/FRET (best with Argon laser)
• Spectral imaging/dye unmixing
• Co-localization (with apochromatic lenses: 40×/1.1, 63×/1.4, or 100×/1.4)

Sample preparation

For imaging with lenses 40× – 100× the samples should be on glass coverslips (e.g. slides, 35mm petri dishes with glass bottom, 40mm Warner Instruments chambers with glass coverslips) or in imaging-quality multi-well dishes. Dry lenses (10×/0.3; W.D. 5.2 mm, 20×/0.5; W.D. 2 mm) can also be used with samples on thicker substrates. No special requirements for fluorescent dyes.

Zeiss LSM 800

Contact: Tomasz Wegierski

Access: training required

General Description

Confocal microscope on inverted stand with Airyscan detector for scanning with enhanced resolution (140nm in XY, 340 nm in Z). Diode lasers: 405, 488, 561, and 640 nm. Detectors: two internal GaAsP PMTs and a 32-channel Airyscan detector. Lenses: 10×/0.3 (dry), 20×/0.5 (dry), 40×/1.3 (oil), 40×/1.2 (water), 63×/1.4 (oil).  Incubation cage and heating insert with CO2 delivery for live imaging. Software and hardware autofocus. Motorized XY stage. Acquisition software: ZEN 2.6. More advanced experiments can be designed with Experiment Designer.

Applications

• Confocal imaging of fluorescently labeled specimens
• Imaging at enhanced resolution (Airyscanning)
• Co-localization (with two apochromatic lenses: 40×/1.2 and 63×/1.4)
• Live imaging (water-immersion lens 40×/1.2)
• Spectral imaging

Sample preparation

For imaging with lenses 40× – 63× the samples should be on glass coverslips (e.g. slides, 35mm petri dishes with glass bottom, 40mm Warner Instruments chambers with glass coverslips) or in imaging-quality multi-well dishes. Dry lenses (10×/0.3; W.D. 5.2 mm, 20×/0.5; W.D. 2 mm) can also be used with samples on thicker substrates. No special requirements for fluorescent dyes (including Airyscanning).