Optical Microscopy

Staff

Core Director:
Lee Cohen-Gould, MS, CEMT
St. Staff Associate in Biochemistry and Cell & Developmental Biology

Multiphoton Facility Director:
Sushmita Mukherjee, PhD, MS
Assistant Professor of Biochemistry

Technologist:
Joshua Sterling, BS

Location:
LC-207, E-135, E-002 WCMC
Phone
212-746-6146 (LCG)
212-746-6495 (SM)
212-746-6199 (MPM lab)

Email:
Lee Cohen-Gould – lcgould@med.cornell.edu
Sushmita Mukherjee – smukherj@med.cornell.edu
Joshua Sterling – jos2048@med.cornell.edu

Research Support
http://www.cornellbiochem.org

Services

  • Consultation. Consultation on experimental design and approach is provided by the Facility Directors.
  • Confocal Microcopy Imaging. The equipment in this facility includes a confocal microscope to be used for 3D image acquisition, and Dell workstations to quantify, manipulate and render 3D fluorescence microscopy images.
  • Training: The Director and Technologist will provide training in usage of the confocal microscope, the Multiphoton microscope and the workstations. Training sessions are by appointment.

Instrumentation

LSM510

  • Upgraded in the summer of 2010, consists of:
  • AxioObserver microscope stand
  • AxioCam MRm digital camera for capturing widefield images that may be incorporated into a confocal image stack (enabling imaging of DAPI-labeled nuclei for example),
  • New computer running both the LSM510 AIM software and Zeiss’s new, ZEN (Zeiss Enhanced Navigation) software.
  • Laser excitations at 458, 488, 514, 543 and 633 nm.
LSM510

Axiovert 200 Wide Field Microscope

  • Controlled with MetaMorph software (Molecular Devices).
  • Lens magnifications ranging from 5x to 63x
  • Brightfield imaging in Phase and Differential Interference Contrast Fluorescence filter sets for DAPI, GFP, CyanFP, Calcium Green, Rhodamine, and CY5.
Axiovert 200 Wide Field Microscope

MultiPhoton Microscopy

The Multiphoton Imaging Facility is a sub-core of the Optical Core facility. It is housed in a dedicated single-use laboratory.

Multiphoton microscopy (MPM) is a nonlinear optical technique that utilizes femtosecond-pulsed near-infrared light as the illumination source. MPM can be used both for fluorescence imaging (tissue autofluorescence, cells expressing fluorescent proteins, as well as exogenous contrast agents), and for imaging utilizing a higher order scattering phenomenon termed Second Harmonic Generation (SHG, generated by oriented non-centrosymmetric tissue components such as collagen and oriented microtubules).

Some advantages of MPM over conventional single-photon imaging include:

  • greater imaging depth (0.5 mm or more in tissue)
  • imaging without the use of exogenous contrast agents (using tissue autofluorescence and SHG signals alone)
  • minimal photodamage to tissue, allowing high-resolution intravital imaging, including studies requiring repeated imaging.

MPM-Ithaca built

MPM-Ithaca built
  • Custom-built for us by the Developmental Resource for Biophysical Imaging Opto-Electronics (DRBIO) at the Ithaca campus of Cornell.
  • Excitation source is a femtosecond pulsed Ti-Sapphire laser (Mai Tai HP, Spectra-Physics, Mountain View, CA), tunable from 700 to 1020 nm.
  • Dedicated to a joint project between the two campuses.

Olympus FluoView FV1000MPE MPM

Installed in the summer of 2010. Purchased with funds from an NIH Shared Instrumentation Grant (Fall 2010).

MPM-Ithaca built
  • Adjustable height Prior stage
  • Excitation source is a Mai Tai DeepSee, with automated dispersion compensation, and is tunable between 690 - 1040 nm.
  • Four non-descanned detectors
  • 405 nm UV laser, (for uncaging and photoactivation, in conjunction with Multiphoton imaging.)
  • Standard objectives
  • IR-transmitting objectives for MPM
  • Two Olympus microprobe objectives (6x and 20x; tip diameter: 1.3 mm).

Olympus stereomicroscope with fluor. & camera

  • Olympus SZX16, with zoom up to 16X.
  • Transmitted light and fluorescence illumination
  • DP20-5E digital color camera

Animal anesthesia and surgery (for intravital imaging)

  • Mouse/rat anesthesia equipment (inhalation anesthesia using isofluorane from VetEquip);
  • Dissection microscope (see above) for microsurgery
  • Carbon dioxide euthanasia setup for mice and rats
  • Mouse pulse oxymeter
  • Basic set of veterinary surgical tools and supplies.

Histology Microscope

  • Olympus BX41 wide field inverted microscope
  • DP20-5E digital camera (shared with the dissection microscope)

3 Image analysis workstations

Software available:

  • MetaMoprh (Molecular Devices),
  • AutoQuant Media Cybernetics)
  • CS3 &CS4 (Adobe)
  • Microsoft Office
  • IQBase (an image management database from Media Cybernetics)