NACH OBEN

Instruments
Zeiss Elyra PS.1 + LSM 880

Type: Confocal Laserscanning and Structured illumination microscope

Laser Lines: 405 nm Diodelaser (30 mW), 458, 488, 514 nm Argonlaser MultiLine (25 mW), 561 nm, 633 nm

Objectives: 10x (Plan-Neofluar 0.3), 20x (Plan-Apochromat 0.8 M27), 63x (Plan-Apochromat 1.4 OIL DIC) and 100x (Plan-Apochromat 1.46 OIL DIC) Objectives

Detectors: Scanhead with 32 channel GaSsP PMT plus 2 channel MA-PMT

Add-ons: Live-Cell incubation unit with integrated temperature and CO2 control, Definite Focus system, Motorized XYZ Piezo-Stage for tiled/multi-position imaging

Software: Zeiss Zen Black

Zeiss Elyra7

Type: Lattice Structured illumination microscope (Lattice-SIM)

Laser Lines: 488 nm (100 mW), 561 nm (100 mW) and 647 nm (100 mW)

Objectives: 10x (Plan-Neofluar 0.3), 40x (C-Apochromat 1.2 W Korr M27) and 63x (Plan-Apochromat 1.4 OIL DIC)

Detectors: 2x pco.edge sCMOS camera with high Quantum efficiency (effective pixels: 1280 × 1280; pixel size 6.5 μm × 6.5 μm; QE: 82%; dynamic range 15 bit)

Add-ons: 70 nm resolution upgrade (SIM2), Live-Cell incubation unit with integrated temperature and CO2 control, 2 Laser-Line Duo-Link System, Definite Focus 2 system, Motorized XYZ Piezo-Stage for tiled/multi-position imaging, Optogenetics upgrade Software: Zeiss Zen Black

Nanolive CX-A

Type: Holotomographic microscope

Laser Lines: 488 nm, FITC, TRITC and CY5 channels additional fluorescence

Objectives: 40x only

Detectors: proprietary Nanolive detector

Add-ons: Live-Cell incubation unit with integrated temperature and CO2 control, Motorized Stage for tiled imaging, automated imaging platform (35 mm and special 96-well plates)

Software: Quantification Software: EVE Analytics, Continuous, multiplexed live cell data, multi-parametric readout

Cytation 5 MPW

Type: Multi-Mode Microplate Imaging Reader

Laser Lines: Scientific-Quad Monochromator with flexible Wavelength for Absorption (230-999 nm) and Fluorescence (230-850 nm)

Fluorescent Channels: DAPI, FITC, TRITC

Objectives: 1.25x and 20x

Detectors: Scientific-Quad Monochromator with flexible Wavelength for Absorption (230-999 nm) and Fluorescence (230-850 nm)

Add-ons: Live-Cell incubation unit with integrated temperature and CO2 control, 2-channel sample injector, High-contrast Brightfield Kit with Differential Interference Contrast (DIC)

Software: Gen5 software for Image acquisition and analysis modules

Sapphire Biomolecular Imager

Type: Laser-based Biomolecular Imager

Laser Lines: 520, 658 and 784 nm

Objectives: 1.25x and 20x

Detectors: Resolution down to 10 µm

Add-ons: Low resolution fluorescent slide-scanner option, Adjustable focus with 7 mm z-sectioning, Chemiluminescent Imaging, In-cell Western Blots, Protein Arrays, In Vivo Imaging

Software: Azure Image capture and Analysis software


Imaging Methods

Laser scanning microscopy

  • Intraorganellar calcium imaging
  • FRAP (fluorescent recovery after photobleaching)
  • Imaging of photoconvertible proteins
  • Spectral imaging and linear unmixing (lambda mode) via 32-channel GaAsP detector
  • FRET (fluorescent resonance energy transfer)
  • FCS (fluorescence correlation spectroscopy)
  • TIRF (total internal reflection) microscopy
  • Proximity Ligation Assay (PLA)
  • Pan-Expansion microscopy (Pan-ExM)

Structured illumination microscopy (SIM)

  • Axial resolution up to 150 nm (Elyra PS.1)
  • 70 nm resolution upgrade (SIM2)
  • Ultra fast and gentle (up to 255 frames per second)
  • Optogenetics upgrade

PALM/dSTORM: Photoactivated localization microscopy

  • Axial resolution up to 20 nm
  • Final image reconstruction from approx. 5000-10000 single images

Holotomographic microscopy

  • label-free single cell and population imaging based on the Refractive Index (RI)

Image Analysis

Imaris 10.1.0 Image analysis software

”State of the art” image analysis. Single full version includes:

  • Segmentation
  • Surface reconstruction
  • Spot/Particle tracking
  • Filament reconstruction
  • Machine learning

CellProfiler 4.2.6
  • Freeware under the GLPv2 public license developed by the NIH and the Broad Institute
  • pipeline/module-based system
  • modules for identification of different cell properties
  • system is able to compute several module outputs with each other