Live Cell and Confocal Imaging Systems

Zeiss LSM 510 META Confocal Scanning Laser Microscope

Description- The Zeiss LSM 510 META CSLM is a state-of-the-art single photon spectral imaging system. The IRF system is equipped with:

  • UV, Argon, green HeNe and red HeNe lasers
  • 3 photomultiplier tubes
  • META detector
  • Transmitted light and DIC capability
  • Zeiss Axioplan 2 motorized microscope
  • Plan Neo 2.5X, 10X, 40X oil, PlanApo 20X and 63X oil, 40X water immersion objectives
  • Software packages for physiology, FRET, FRAP and 3D
  • Additional workstation with full software packages for offline image processing

Applications-

  • Imaging of specimens labeled with multiple fluorochromes from the UV through far red ranges
  • Measurement of alterations in ion concentrations
  • Visualization of protein interactions
  • Determine protein kinetics
  • Quantitative co-localization studies
  • Excitation and emission fingerprinting of fluorochromes
  • Multichannel unmixing
  • 3D reconstructions

Location- The LSM 510 system is located in Building 1 Room B60.

Website- www.zeiss.com

Zeiss VivaTome Spinning Disk Confocal Live Cell Imaging System

Description — The Zeiss VivaTome system is a spinning disk confocal system mounted on a Zeiss Axiovert  200M inverted microscope stand. The system offers the ability to monitor rapidly occurring events within living cells without compromising resolution while employing high frequency low intensity illumination that substantially reduces photobleaching and phototoxicity. This in-vitro system allows for real-time observation and image recording of live cells by signal collected by phase contrast and fluorescence optics. The IRF system consists of:

  • Illumination Sources
    • High intensity HAL 100 source for brightfield and phase contrast imaging.
    • Exfo X-Cite 120 Metal Halide high intensity source providing wavelengths from 360-700nm at constant power across the illumination field.

Optics: Phase contrast and brightfield objectives:

  • 10x NA 0.25 PH-1 Achroplan
  • 20x NA 0.40 PH-2 Achroplan
  • 40x NA 0.75 PH-2 Achroplan

Optics: Fluorescence objectives

  • 10x NA 0.5 FLUAR
  • 20x NA 0.75 FLUAR
  • 40x oil NA 1.3 FLUAR
  • 63x oil NA 1.4 Apochromatic

Filters and Dichroics

  • 490/515 FITC, Cy2
  • 555/570 TRITC,Cy3,Texas Red
  • 635/670 Cy5
  • 340-360/510 Fura-2 for ratiometric calcium imaging
  • 492/520 for GFP imaging
  • 360/460 for DAPI, Hoechst and AMCA imaging

Image Capture

  • Image capture is by a Hamamatsu ORCA-ER digital camera controlled by Kinetic Imaging Image Acquisition Software (AQM Advance-6)

Software Packages

  • Zeiss Vivatome image analysis package
Environmental Control
  • The environment of the microscope stage and specimen under observation is by a Zeiss M 200 Incubator chamber and heated stage and objective controllers. This system allows for the control and stabilization of the temperature, CO2 concentration and humidity.

Specimen holders

  • The microscope stage accommodates standard slides, Lab-Tek chambers, 35-60mm Petri Dishes, Microplates, and small, specialized culture chambers.

Applications

  • Long-term time-lapse studies including image capture in phase contrast for investigation of cell migration patterns and cell movements under normal and experimental conditions.
  • Monitoring of cell division.
  • Short term, high speed imaging (up to 30 frames/sec) for imaging of ion flux in living cells, in particular +calcium, sodium and pH.

Location- The AttoFluor CARV-1 Confocal Live Cell Imaging System is located in Building 4 Room B24 in the School of Medicine complex.

IRF Contact: John Fuseler

Website- www.bdbiosciences.com