WILLIAMS, GEOFFREY L.* AND KAREN L. KLOMPARENS. Center for Electron Optics and Department of Botany and Plant Pathology, Michigan State University, MI 48824. - Picea pungens pollen, a three dimensional exploration.
The Laser Scanning Microscope (LSM) provides a tool for studying
morphology in a totally new way, in three dimensions, in various
conformations and also in motion. Additionally, the LSM can further
enhance visual comprehension by showing the inside and the outside of
a cell at the same time. Small, optically transparent tissues are
best for this type of exploration, and many investigators are
beginning to use this as an important tool in their work. I am using
spruce pollen to exploit this new technology, as the pollen grains are
on the large side of the high-resolution limit. The size of the
pollen grains I examined are around 60 microns, within the limits of
the working distance on a high NA objective, but too big for
traditional mounting techniques. The resulting high-resolution images
generated by the Silicon Graphics, Inc. workstation (SGI) provide
similar surface detail to the scanning electron microscope and much
improved detail over a conventional fluorescence light microscope.
The internal information is limited to the fluorescent stain used and
the laser lines available on the LSM. The samples were stained with
Acridine Orange and excited with a 488nm laser. The techniques used
are quick and easy for pollen, as it can be collected fresh and
stained or collected, fixed, then stained and mounted at a later date.
The SGI helps to create 3-D models that are easy to understand and
visually illustrate more information at once without using cartoons or
the imagination. The information gleaned from LSM and SGI processed
images combines high surface detail with clear internal information
that is equivalent to more time consuming techniques thus speeding up
information gathering on the morphology of pollen in a format amenable
to presentation in the computer age.
Key words: 3D-reconstruction, LSM, Picea, pollen, SGI