Funding from Indiana University, the NIH and the Lilly Endowment have given Indiana University a wor

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Indiana Center For Biological Microscopy
RESEARCH CONTRIBUTION HIGHLIGHTS
LIST OF SERVICES
OVERVIEW

• The Imaging Facility provides researchers with
  training and access to advanced optical
  microscopy equipment, both for image collection
  and for image analysis.
• Facility personnel provide consultation on the
  design and analysis of microscopy studies.
• The Facility is active in developing new
  methods of microscopy and digital image analysis,
  including developing and optimizing multiphoton
  microscopy, particularly for studies of living
  animals and developing novel methods of digital
  image analysis.
• The Facility is active in education, providing
  frequent seminars on microscopy, participating in
  the graduate program on Biophysical Imaging, the
  undergraduate Biotechnology training program and
  a national course for renal researchers.
• The Facility maintains a website that in
  addition to supporting on-line equipment
  scheduling, also provides numerous tutorials on
  subjects such as Fluorescence Resonance Energy
  Transfer, microscopy of living cells and
  quantification fluorescence co-localization.

Funding from Indiana University, the NIH and the
Lilly Endowment have given Indiana University a
world-class center for biomedical imaging. The
Indiana Center for Biological Microscopy (ICBM)
is one of a handful in the world equipped for
low-light level microscopy, confocal microscopy,
UV confocal microscopy, 2-photon microscopy,
digital deconvolution microscopy, live cell
microscopy and the latest systems for digital
image analysis and visualization. The
facility represents a strong institutional
commitment to optical imaging technology
development. Since 1998, Indiana University has
committed more than 5M to this facility.
In addition to providing state-of-the-art support
for School of Medicine members, the core is also
actively involved in research into biological
imaging, resulting in the development and
dissemination of new methods of microscopy and
digital image analysis software. One of the
products of this research is the ongoing funding
of the Indiana University OBrien Center, whose
mission is to develop advanced methods of
intravital and 3-dimensional microscopy. The
products of these activities are disseminated
through a program of education, including
seminars, courses and individual training.
Volume rendering of a fluorescence microscopy
volume of the middle of gastrulating mouse
embryo, 7 days into development. The earliest
hematopoietic cells (blue) are just emerging from
the primitive streak. Michael Ferkowicz and Merv
Yoder are looking at these progenitor cells to
better understand the proliferation and
differentiation of hematopoietic cells during
development.
3-dimensional fluorescence image of a colony of
endothelial progenitor cells. Nuclei of all
cells are labeled with blue Hoechst nuclear
stain, and uptake of red acetyl-LDL is used to
distinguish early progenitors (center cluster of
cells lacking LDL uptake) from spindle cells
radiating out from the center and endothelial
cells, both of which take up the red fluorescent
acetyl-LDL. Daniel Prater and David Ingram are
using this cell system to better understand the
development of endothelial cells.
QUALITY CONTROL AND ASSURANCES
RESOURCES

• Facility personnel perform rigorous regularly
  scheduled testing to ensure optimal performance,
  and modify systems according to users needs.
• This includes
• Point Spread functions for testing light source
  alignments and chromatic and spatial aberration
  of objectives.
• Laser power is monitored with a power meter at
  the plane of the back aperture.
• Daily Images with standard slides to test image
  quality.

• Bio-Rad MRC1024 laser scanning confoca and
  multiphoton microscope is equipped with a
  Krypton-Argon (488, 568, 647 nm excitations)
  laser for confocal microscopy and a tunable
  Titanium-Sapphire laser (using a 5W Millennia
  diode solid state pump laser) for multiphoton
  microscopy.
• Perkin-Elmer UltraVIEW spinning disk confocal
  system equipped with 3 lasers providing
  excitations at 442 nm, 488nm, 514 nm, 568 nm and
  647 nm.
• Zeiss LSM-510 confocal microscope is equipped
  with a UV Argon Laser (351 nm, 364 nm
  excitation), a visible Argon laser (458, 488,
  514nm excitation) and two Helium-Neon Lasers (543
  nm and 633 nm excitation).
• Zeiss LSM510-Meta confocal and multiphoton
  microscope system equipped with an Argon laser
  (458, 488, 514nm excitation) and two Helium-Neon
  Lasers (543 nm and 633 nm excitation) for
  confocal microscopy, and a tunable
  Titanium-Sapphire laser (using a 10W Millennia
  diode solid state pump laser) for multiphoton
  microscopy.
• Inverted Nikon wide-field epifluorescence digital
  microscope equipped with a Hamamatsu Orca cooled
  CCD detector.
• Upright and inverted Nikon wide-field
  epifluorescence microscopes equipped with SPOT
  cooled color CCD detectors.
• An Eppendorf micromanipulator and microinjector
  is available for microinjection studies of living
  cells
• Two Hewlett-Packard workstations devoted to image
  analysis.
• All Microscope Systems and Analysis computers are
  connected over a 1000Base-T network.
• Our staff is an invaluable part of the services
  that we provide.

Intravital multiphoton fluorescence imaging of
the bone marrow of the calvarium of a transgenic
mouse that expresses GFP in bone marrow
hematopoietic progenitors. The vasculature was
visualized using a Texas-Red dextran, injected IV
5 minutes prior to imaging. This study is part of
a 13 investigator pilot project, led by Nadia
Carlesso and Ken Dunn, aimed at developing
intravital microscopy as a tool for Cancer Center
investigators to study the cancer
micro-environment in vivo. This project will
culminate in an intravital microscopy service
core for the Cancer Center.
CONTACT INFORMATION
Core Director Kenneth W. Dunn, Ph.D. Address
Research Institute II, 950 West Walnut
St, E233 and E243 Website
http//www.nephrology.iupui.edu/imaging/
Intravital multiphoton fluorescence imaging of
filtration in the glomerulus of a living rat.
Nucei are visualized with Heochsts (blue). A
texas-red dextran was injected after acquisition
of frame A. In B the dextran is being filtered
from the blood and can be seen within the
Bowmans space of the glomerulus. In C the
dextran has been concentrated and appears bright
in the distal tubules. In D the dextran has been
excreted. This study is part of the OBrien
Center for Advanced Renal Microscopic Analysis,
whose primary goal is to develop new optical
methodologies for investigators in Nephrologic
and Urologic Research.