Confocal-Microscopy-Core.jpg
Bookmark and Share

Live Imaging Laboratory (LSM 5 Duo)

Capabilities

The Live Imaging Laboratory Supports:

  • Fast imaging of live preparations (detailed motion studies)
  • Separation of up to four fluorophores in a single sample (using LSM510)
  • Fluorescence excitation of CFP, GFP, fluorescein, Fluo-3, YFP, rhodamine, Cy5 and DAPI on the LSM510 and CFP/GFP Rhodamine on the LSM 5-Live.
  • Forster resonance energy transfer (FRET) and anisotropy-FRET studies
  • Automated collection of optical sections, for three-dimensional reconstruction
  • Automated collection of time series, for studies of dynamic processes and fluorescence recovery after photobleaching (FRAP)
  • Lateral resolution of up to 250nm, axial resolution of up to 1 um
  • Environmental chamber for temperature and atmosphere control for long-term studies of living samples
  • Laser scanning Nomarski differential interference contrast transmission microscopy

Hardware

  • AxioOberver Z1 inverted microscope
  • Objectives: 63x/NA1.4 oil immersion plan-apochromat, 40x/NA 1.3 oil immersion plan-neofluor, 40x/NA1.2 water immersion C-apochromat, and 20x/NA0.8 multi-immersion plan-neofluor. 10x/NA0.3 plan-neofluor also available.
  • Long working distance condensor (NA=0.55)
  • Special optics for Anisotropy-FRET applications
  • DIC Optics
  • DC motor with optoelectronic coding, smallest increment 25 or 50 nm. Fast piezoelectric focusing drive acting on 63x objective.
  • Motor-driven XY scanning stage with Mark&Find and Tile Scan (Mosaic Scan) functions; smallest increment 1 μm
  • AxioCam Digital Microscope Camera

Zeiss LSM510 META laser scan head 

  • 13 x 2 speed stages; up to 5 frames/s with 512x512 pixels (max. 77 frames/s with 512x32 pixels), 0.38 ms for a line of 512 pixels
  • Omnichrome Argon ion laser
        ~25mW of 458nm, 488nm, and 514nm excitation light
  • Zeiss DPSS with 10 mW 561 nm excitation light
  • Zeiss Helium/Neon laser with 5mW 633nm excitation light
  • Zeiss Diode laser with 30 mW of 405 nm excitation light
  • Three fluorescence Hamamatsu photomultiplier tube detectors + Transmission PMT
  • Dichroic mirrors and filters for simultaneous or sequential observations of AMCA/Indo-1, CFP, fluorescein/fluo-3/GFP, YFP, rhodamine/RFP and Cy5 fluorophores, and for reflection confocal microscopy
  • Transmitted light detector

Zeiss LSM5 Duo laser scan head 

  • up to 120 frames/s with 512x512 pixels; faster modes with smaller frames (e.g. 505 frames/s with 512x100 pixels, 1010 frames/s with 512x50 pixels) ultrafast line scan mode with >60,000 lines/s
  • Simultaneously image two channels. Equipped with highly sensitive detectors (QE 70% or better). Bright-field transmitted-light mode possible.
  • Zeiss diode laser with 16mW of 440 nm excitation light
  • Zeiss diode laser with 100 mW of 488 nm excitation light
  • Zeiss diode laser with 75 mW of 532 nm excitation light

Location

The Live Imaging Laboratory is located in space provided by the Department of Physiology on the fifth floor of the Bressler Research Building, 655 W. Baltimore St, Room 5-042.

Funding Sources

NSF Grant for the Hyperspectral Confocal Microscope

In September, 2004 the Division of Biological Instrumentation and Resources of the National Science Foundation provided $346,692 to establish the Hyperspectral Confocal Imaging Laboratory. The grant was organized by P.Luther, (Physiology), along with co-PIs B. Alger (Physiology), R. Bloch (Physiology), W. Lederer (UMBI), W. Randall (Pharmacology), M. Schneider (Biochemistry), S. Thompson (Physiology), T. Rogers (Biochemistry), J. Wade (Physiology), and E. Weinmann (Nephrology). The University of Maryland School of Medicine and Department of Physiology contributed a total of $148,582 in matching funds.