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VayTek offers Advanced Image Processing Systems, including both hardware and software for:

• Microscopy
• Industrial Inspection
• Medical Imaging
• Quality Control
• Non-Destructive Testing
• Deconvolution of Confocal Images
• 3D Volume Visualization and Measurement

VayTek, Inc.
505 North Third Street
Suite 200
Fairfield, IA 52556
Tel 641-472-2227
Fax 641-472-8131
Email vaytek@vaytek.com

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Application Note

Uses for VayTek's Software in Study of Alzheimer's Disease

Richard Kraig, Ph.D., et al, have imaged microglial cells within a hippocampal organ culture. Out-of-focus haze was removed with VayTek's MicroTome software, and the 3D projection was made using VayTek's VoxBlast software.

Richard Kraig, Ph.D.

This before/after image appeared on the cover of Microscopy & Analysis, September, 1997. It shows a microglial cell stained with IsoB4Lectin (Sigma) within a hippocampal organ culture. The culture is 100-150 microns thick and closely mimics its in vivo counterpart. Individual cells within this isolated culture system can be followed in space and time, making it an ideal preparation to examine cellular and molecular changes within normal and diseased brain tissue.

This image was captured with a Photometrics CH250 camera. The right half of the image was processed with a digital deconvolution program, MicroTome, available from VayTek, using a theoretical point spread function. 68 slices were captured at .18 microns per step at 100x oil objective. From these slices a 3D projection was made using VoxBlast, a 3D reconstruction program from VayTek. The use of deconvolution and 3D reconstruction programs allows for increased accuracy in the measurement and visualisation of the microglial cells.

This research relates to answering larger questions that include; what role astrocytes and microglia have in making, maintaining and modifying synapses; how these cells influence the evolution of ischemic brain injury; how they regulate the capacity for regeneration after ischemic injury; and how glia may participate in the development of inflammation from Alzheimer's disease.

Whole animals, primary culture of astrocytes and microglia as well as hippocampal organotypic cultures are used to examine cellular behavior. The fact that brain cells do not exist in isolation is stressed. Accordingly, much effort is directed toward considering how glia interact with each other and with neuronal cells to affect brain function. A variety of electrophysiologic tools (including ion-selective, DC, and patch clamp electrodes as well as ratiometric imaging strategies) are used to monitor functional changes in cells.

How changes in membrane potential, pH, calcium and neurotransmitters interrelate to effect physiological, biochemical and molecular changes in glia and in their adjacent neuronal counterparts are studied. Imaging techniques (including quantitative immunohistochemistry, computer optical sectioning microscopy and digital deconvolution, and 4-dimensional reconstruction of dye filled cells) are used to assess changes in shape and function of glia.

Culture prepared by Dr. Phillip Kunkier; Image processing by Raymond Hulse; Richard P. Kraig, Ph.D./M.D., Laboratory Director; Cerebrovascular Disease and Aging Laboratories; Department of Neurology, University of Chicago