Equipment/ Instruments

The Two-Photon Imaging Facility is equipped with two multi-photon microscope systems:

  1. LaVision BioTec TriMScope I
  2. Olympus FV1000MPE

    and two in-vivo optical imaging systems:
  3. Perkin Elmer IVIS Lumina XR
  4. Berthold NightOWL

LaVision BioTec TriMScope I

Copyright: 2PIF

Microscope table: Anti-vibration table, active dampening, Newport ST-series
Laser:

Spectra Physics MaiTai HP

Laser modulation: LaVision BioTec TriMScope I with fixed pre-chirp, optical beam multiplicator and non-resonance mirror scan head
Microscope stand: BX61WIF (motorized upright microscope)
Detection: Single beam mode: filter based detection system; imaging with up to six detectors is possible (three direct PMT detectors (d=25 mm) at the normal PMT port and three GaAsP detectors (Hamamatsu H 7422-40) at the ultrasensitive PMT port with optional exchange of detectors between the ports). One detector port in transmission below the sample stage (usually not occupied). Spectral filtering is performed by manual selection of dichroic mirrors and bandwidth filters in front of each PMT.
Multi-beam mode:

Two highly sensitive Hamamatsu C9100-02 EMCCD cameras with motorized filter wheel (LaVision BioTec FW8-0036) for spectral filtering.

Fluorescent filters available from 380-670 nm in approx. 20 nm and 50 nm bandwidth steps (including corresponding dichroic mirrors).

Microscope stage: Motorized LaVision BioTec intravital X/Y stage.
Incubation system: Life imaging services incubation system CUBE&BOX. Sensor: Pt-100 thin- layer RTD temperature range: +5 C to 50C Stability: 0.1C at sensor location, Precision: 0.3C at sensor location
Image acquisition computer: Windows 7 Prof. SP1 64Bit system; Intel i7-3770 CPU @3,40/3,41 GHz, 3,19 GB RAM, Nvidia Quadro 410, 30-inch flat screen, LaVision TriMScope control/trigger board, second AD-Board and Hamamatsu C9100 control board.
Image acquisition software: LaVision BioTec Inspector Pro
Others: Classic HBO fluorescent light source (X-Cite120) including light guide, four fluorescent filter cubes, bright field trans-illumination light source (TH4-200) with an aplanatic achromatic 1.4 condenser. Laser safety glasses and IR-viewer.



Olympus FV1000MPE

Microscope table: Anti-vibration table (active dampening) (Newport RS2000-series)
Laser: Spectra physics MaiTai DeepSee HPDS-OL with wavelength adjustable pre-chirp. Argon laser diode (458/488/515 nm), including a multi-laser combiner unit.
Laser beam modulation: Olympus Fluoview FV10 single beam scanner optimized for NIR wavelengths.
Microscope stand: BX61WIF (motorized upright microscope)
Detection:

Filter-based detection system; three internal non-directly scanned PMT detectors (d=25mm). Bandwidth selection of emitted light by filters in motorized filter wheel is in front of each PMT (confocal detectors). Four ultra-sensitive external non-descanned PMT detectors (d=32mm) for direct detection of emitted light. Bandwidth selection by spectral filters and corresponding dichroic mirrors (in a filter cube) in front of each PMT. Internal optical zoom unit.

Microscope stage: Motorized Prior Z-Deck X/Y stage.
Image acquisition computer:

Windows 7 Prof. 64bit system; Intel Xeon CPU E5-1620-0 @ 3,60GHz/3,60GHz, 8 GB RAM with FluoView interface board, 30 inch TFT- monitor.

Others:

Classic HBO fluorescent light source including light guide, two fluorescent filter cubes. CCD camera Olympus XM10 (12 bit monochrome camera, serial #: 51000191) with Cell-P acquisition software. Laser safety glasses and IR-viewer.

Objectives: XLUMPLAN20x
XLPLN25xWMP-SP
LUMFI60x
UMPLFLN 10XW
XLPLN10XSVMP
20x
25x
60x
10x
10x
NA 0.95
NA 1.05
NA 1.10
NA 0.30
NA 0.60
WD2.0
WD2.0
WD2.0
WD3.5
WD8.0

Additional equipment:

  • Small animal surgery station including stereomicroscope and S1-lab bench with basic equipment for preparation of chemicals.
  • Trigger box for the transformation of external signals (ECG and respiration) into trigger pulses for imaging.
  • 1st image processing computer: “Datec 1” (64 GB RAM)
  • 2nd image processing computer: “Datec 2” (192 GB RAM)
  • 3rd image processing computer: “Fujitsu R640” (192 GB RAM)

Image processing software:

  • ImagePro Analyzer 3D 7.0
  • Bitplane Imaris
  • Fiji (ImageJ)
  • Autoquant X3
  • Olympus Fluoview FV10 2.0 viewer
  • LaVision Biotec Inspector Pro

IVIS Lumina XR

Frontview of the IVIS Lumina XR Copyright: 2PIF
Light source: Halogen Tungsten Lamp (150 W), LED lamps for photographic images.
Detection: -90C cooled back-thinned, back-illuminated grade 1 CCD camera, 1024x1024 pixel, 13x13mm, (16Bit).
Filters: 10 position excitation filter wheels, equipped with 10 excitation filter (30nm bandwidth) in the range from 430-745 nm, 8 position emission filter wheel, equipped with 4 emission filters (GFP/DsRed/Cy5.5/ICG). Automated image integration to overlay with bioluminescence, fluorescence and photograph.
Imaging chamber: Light-sealed, heated stage to maintain optimum body temperature. Motor controlled stage, filter wheel, lens position, and f-stop.
X-Ray module: Radiation shielded cabinet. Fast X-ray image acquisition times of 1-10 s reducing radiation exposure.
Additional accessories: Optical zoom lens attachment for close up and high resolution X-ray images, optical X-FOV lens attachment for view field extension to 23x24 cm (5 mice). Gas anesthesia ports, 5 position manifold within imaging chamber. Syringe injection system
Anesthesia snit: Xenogen XGI-8 gas anesthesia system for small animals (adapted to anesthetize up to five mice in parallel).
Software: Living Image Software 4.4

Berthold NightOWL

Frontalview of the Berthold NightOWL Copyright: 2PIF

Light source:

Halogen 75 W tungsten lamp, ring-light fluorescent illumination.

Detection:

Andor Ikon, -70C cooled back-thinned, back-illuminated grade 1 CCD camera, 1024x1024 pixel, 13x13mm, (16Bit).

Filters:

4 excitation filters per slide. 4 emission filters per wheel.

Imaging chamber:

Light-sealed cabinet with motor-driven camera. Sample size up to 250 mm. Temperature controlled animal tray

Anesthesia unit:

TEM gas anesthesia system for small animals (adapted to anesthetize up to five mice in parallel).
Software: IndiGOTM software