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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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VayTek's Digital Deconvolution Imaging System was used to view biofilm organisms.

The Biotechnology Research Department at the Orange County Water District has constructed a digitally-based microscope imaging system capable of high-magnification imaging of microbial biofilms and subsequent rendering of those images in 3D virtual space. By staining the biofilm organisms with fluorochromes reacting to changes in physiological status, the effects of various agents on microbial components of the biofilms may be investigated. The system consists of a standard light microscope to which several optoelectonic components are fitted that allow automatic optical sectioning of microscopic specimens, followed by digital removal of defocused data, reassembly of the sections and display of the reconstructed specimen in 3D. This presents the microscopist with a "virtual reality" environment in which to examine a vastly enlarged and enhanced representation of the original specimen.

The images are initially obtained using a light-microscope equipped with oil immersion UV and standard epifluorescent illumination optics (Olympus IX 70 Inverted Microscope). All optical components are standard, except that a 1 joule short arc xenon flash (EG and Optoelectronics, emission 256-10000nm) is used rather than a continuous wave mercury lamp as the source of excitation light. This minimizes fluorochrome photobleaching and allows a quantitative determination of fluorescence.

A Ludl drive motor attached to the microscope stage position knob permits acquisition of computer-controlled serial optical sections. Depth of field consideration currently limit practical z-axis resolution to 0.68 microns.

Projection eyepieces ranging from 1.67 to 6.7x are used in the microscope trinocular head to convey images to specially modified (manual gain added) 3rd generation image intensifier (Dark Invader Owl, B.E. Meyers and Co., Redmond, WA). The images are captured using a cooled CCD camera (Dage IFG 300). The camera uses a standard RS170 link to transfer data to a digital capture board in a Pentium-based PC (with an image capture board from Integral Techologies, Inc., Flash Point 128.) Digital image size is 512 x 480 pixels, which with a 2.5x projection eyepiece yield a spacial resolution of 9.227 microns in the x-axis and 0.191 microns in the y-axis. Each image in the image stack (monochrome 8 bit) is processed to filter out noise using ImagePro-Plus, then digitally deconvolved using VayTek's MicroTome AT devonvolution software to remove defocused information.

Once an image stack is processed, it is transmitted via an ethernet connection to a UNIX workstation (Silicon Graphics Iris Indigo Elan 4000, 382Mb RAM). The image stack is assembled into a volume using VoxBlast UNIX. Data are resampled, and interpolation is used to produce a voxel volume accurately representing the correct z-axis aspect of the original specimen. Voxel volume data are typically rendered in false color (24 bit) and false shadow to accentuate contrast of the original grey level image, permitting both manual and machine recognition of features within the rendered volume. The resulting virtual reconstruction may be freely manipulated by the microscopist in all three axes, allowing stereoptic display, virtual dissections, polygon extractions of areas of interest, and both 2 and 3D tomography.

Virtual images created by VoxBlast are recorded by first capturing into a bitmap editor, followed by enhancement to improve appearance and add features (text, borders, size bars, multiple or inset imaging, etc.) Images are then either digitally archived to tape or printed using a dye sublimation printer (Kodak ColorEase) onto either paper or transparency medium.