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Courtesy of John George, PhD in Biophysics and Neuroscience at Los Alamos National Laboratory: Rat brain nerve cell reconstructed from a series of confocal slice images collected with a prototype "Virtual Pinhole Microscope" (see press release below). The shadowy area left of the nerve cell is an electrode track.

NEWS RELEASE

VIRTUAL PINHOLE MICROSCOPE

A Revolution in Confocal Microscopy

Los Alamos Laboratory and VayTek, Inc recently announced the development of a prototype for the "Scanning Computed Confocal Imager" or Virtual Pinhole Microscope. This breakthrough technology takes advantage of existing improvements in digital imaging systems, but configures functional subsystems in a novel way that eliminates many of the limitations and much of the expense associated with conventional confocal imagery.

One of the key elements of the new Imager is the use of a "virtual" aperture. The virtual aperture is implemented in software on a host computer, such as a pentium-based computer or a Power Mac. It is possible, with this system, to adjust key parameters (such as effective aperture size) to optimize the image even after the basic data are acquired. Several algorithms have been used to produce images similar to those acquired through conventional confocal microscopy.

A key subsystem of the Virtual Pinhole Microscope offers an elegant solution for providing scanned illumination. John George, PhD in Biophysics and Neuroscience at Los Alamos, uses spatial light modulators (SLMs) to produce time-varying spatial patterns of illumination. The SLM that Dr. George is using on the current prototype is a liquid crystal display (LCDs) similar to those used in some video projectors. By setting the polarization on the LCDs, Dr. George can program light patterns from a single point to a complicated spatial pattern, to one or more lines or slits -- choosing optimal patterns in space and time for capturing the desired images. Other technologies, such as ferroelectric liquid crystals, micro-mirror display systems, or electronic laser scanning systems can be adapted for use with the Scanning Computed Confocal Imager.

The LCD illumination grid is evident in this image. This is one of the images used to construct the confocal image shown at the beginning of the news release. Courtesy John George, PhD, Los Alamos National Laboratory.

Other Features of the Virtual Pinhole Microscope Include:

• No Moving Parts -- provides an excellent basis for a confocal and spectroscopic endoscope system.
• Easy to Retrofit -- adaptable to existing systems, including conventional microscopes with video capability ·
• Low Cost -- high performance, extended sensitivity, speed, and dynamic range from low cost components
• Can be configured to operate in transmitted light modes, reflected light, or epifluorescence modes
• Multiple images with different effective contrast mechanisms can be computed from a single data set
• All attributes of the system are programmable

Examples of Applications for the Virtual Pinhole Microscope Include:

Optical Imaging of Neural Function -- Maximizing the ability to obtain confocal images in real time, researchers could measure changes in the optical properties of the cortex that reflect neuronal activity. Virtual Pinhole Confocal techniques would allow different response properties at different cortical layers to be measured at a much higher resolution. These measurements could describe the spatial response pattern across the cortex at each of the cortical layers and sublayers.

High Speed Imaging -- Although most of the power in macroscopic electrophysiological measurements of neural population activity is at frequencies between 10 and 100 Hz, firing rates of individual neurons may approach 1kHz. Oscillatory population responses are observed in a range of 40-60Hz. It would be useful to resolve phase relationships between active neural populations. Existing technical approaches using slow scan or standard video devices cannot address important temporal dynamic regimes within single cells or networks of cells.

Engineering and Material Science -- The new system offers a rugged, lighter weight imager with greater flexibility and improved reliability.

Inspection and Industrial Quality Control -- Most inspection sites, such as those associated with semi-conductor inspection, have invested heavily in expensive equipment. The ease with which the Virtual Pinhole Microscope can be retrofit to existing systems makes it ideal for these systems.

A complete Virtual Pinhole Microscope system consists of imaging optics (an existing microscope, for example), a system for scanned illumination, a standard or high performance solid state video camera, and a computer system for image acquisition, scan control, and image reconstruction.

Dr. John George and his colleagues at Los Alamos National Laboratory have a patent pending on the system. VayTek, Inc. has contributed expertise in digital imaging techniques and has licensed the Virtual Pinhole Microscope for manufacture and distribution.

To inquire about the Virtual Pinhole Microscope, contact VayTek at 641-472-2227 or vaytek@vaytek.com. For further information on image acquisition and reconstruction visit the VayTek web site at http://www.vaytek.com.

Courtesy VayTek, Inc., Fairfield, IA. This color image consists of three separate 12 bit grayscale images, each captured at a different wavelength for three different dyes: FITC for the actin fibers, rhodamine for a protein, and DAPE for the nucleus. Images were captured at 6 different vertical depths at 1 micron steps. Each color channel was processed separately, and a topview composite image was created. By compositing several images from different vertical depths in the image a more complete view of the specimen is produced. The composite view of the images with the haze removed is much more revealing than a topview of the unprocessed images. See news release below.

NEWS RELEASE

New MicroTome (TM) Software

Digital Imaging Systems for Microscopy

MicroTome, produced by VayTek, Inc. of Iowa, is a software package that operate as part of a digital imaging system for scientific research. MicroTome is ideal for researchers who need to keep costs down, but must have the low-noise, highly detailed images that are normally obtained from expensive confocal equipment. MicroTome is ideal for scientists and engineers who need to remove out-of-focus haze from microscope images.

MicroTome run under Windows and Macintosh operating systems.

MicroTome is more flexible than other deconvolution software. It allows the user to select one of many algorithms, including VayTek's proprietary single image haze removal or constrained iterative algorithms. MicroTome also accepts both measured and calculated point spread functions.

MicroTome is more versatile than similar software. It works with a broader range of fluorescent, transmitted, reflected and other image modes.

In most instances, images deconvolved with MicroTome are as good as those acquired using a confocal microscope -- at less than a quarter of the cost. Even images captured with a scanning confocal can be deconvolved with this software, improving the image resolution.

The MicroTome software package also provids a suite of tools that aid in quantitative analysis and in image processing. Cell counting, densitometry, two dimensional image processing that includes filters, and color image processing are some of the features. With a click of the mouse, MicroTome automatically creates true color, 24 bit (RGB) images.

To inquire about MicroTome, contact VayTek at 641-472-2227 or vaytek@vaytek.com.