Recent developments promise the possibility to externally control every aspect of microscopes through a computer interface. In combination with high-resolution cameras and feedback to the microscope, this can be leveraged to create highly automatic routines, e.g., to remotely correct astigmatism. Together with the development of fast computer networks this creates a new branch of microscopy, the so-called “telemicroscopy”. The goal of telemicroscopy is the control of a microscope over a large distance including the transfer of images with an acceptable repetition rate. A big advantage for electron microscopy in particular is the possibility of having access to expensive and well-equipped microscopes. In the field of light microscopy the branch “telemedicine” was created, meaning the “virtual” presence of a colleague or specialist for discussion or diagnosis.

Using transmission electron microscopy as an example, the history and special requirements for automation and telemicroscopy will be discussed. In the late 80's the first TEM with a remote control was revealed. Shortly thereafter, first automatic functions for defocus control and astigmatism correction were developed using a video camera as electronic image converter.

The field of in-situ electron microscopy has recently become very active. Microscopy and Microanalysis 1997 was host to an extended symposium devoted entirely to in-situ studies. Also, a recent Winter School sponsored by Arizona State University was devoted entirely to in-situ electron microscopy and drew over 90 attendees. Examination of the presentations and papers in these in-situ symposia has revealed that the researchers involved still have problems acquiring, managing and storing and manipulating their data. An in-situ experiment, by its very nature, is dynamic and recording micrographs is typically not the optimum method of image acquisition. Videotape is the preferred medium. High grade S-video recording of experiments permits the extraction of single frames of reasonable quality from the tapes for presentation and publication. There are, however, a number of problems with video recording. During in-situ studies a number of variables in the sample environment are changing, e.g. the pressure, the temperature, the stress, the strain and the microscope parameters (accelerating voltage, beam current, magnification, specimen tilt, etc.).

The Materials Microcharacterization Collaboratory (MMC) was created last year as a pilot project within the US Department of Energy's DOE2000 program. The DOE2000 program has three main goals: improved ability to solve DOE's complex scientific problems, increased R&D productivity and efficiency, and enhanced access to DOE resources by R&D partners. One of the strategies to meet these goals is the construction of national collaboratories. These provide integration on the Internet of unique or expensive DOE research facilities and expertise for remote collaboration, experimentation, production, software development, modeling, or measurement. In addition, collaboratories will benefit researchers by providing tools for video conferencing, shared data-viewing, and collaborative analysis. Cooperative pilots projects, jointly funded by DOE2000 and a scientific program area, are expected to lead to clear success in the scientific environment: they are to provide new capabilities and to increase the efficiency of doing the work (e.g., reduce travel, increase communication, provide for sharing of data among formerly disparate environments).

We are developing a testbed for automated identification of specimens, and acquisition of high resolution images for 3D macromolecular EM. Several software package which have been developed specifically for acquisition of tomographic data sets have illustrated the benefits of automation. These software packages require the user to identify a specimen of interest and then automatically acquire a single tilt series from the identified specimen. The testbed we are developing will provide a generalized system for specimen identification, image acquisition and image quality assessment. The goals of the testbed are to provide a system that will acquire EM images unattended by an operator.

The testbed integrates a computer controlled TEM with a distributed real-time image processing environment. Computerized control and image acquisition from the TEM uses the emScope software package (1). This is coupled to existing image processing software packages and new software tools developed specifically for this project.

The Microscope and Graphic Imaging Center (MAGIC) at California State University, Hayward, has developed a model for interactive remote shared access to visual and analytic scientific instrumentation (IRSA/VASI). Using specialized software over the Internet, instruments such as electron microscopes can be shared throughout the world. Remote access to a centralized microscope provides investigators with specialized equipment that they might not be able to purchase and support on their own. For those who already have such instruments, it affords access to additional equipment and permits them to use their own resources in a wider context. Remote control of instrumentation is being researched and developed at national laboratories, supercomputing centers, and universities. Remote access and control was successfully demonstrated in 19921 and since that time, there have been other major achievements.

To demonstrate our model of interactive remote shared access to instrumentation, we have developed a system of software programs for remote control of a Philips XL-40 scanning electron microscope.

Our original “Telemicroscopy” project has now been expanded into a project in which we aim to provide a “Collaboratory for Microscopic Digital Anatomy” (CMDA) Tools being developed will provide a researcher at a remote site distributed interaction with unique instrumentation for data acquisition, as well as significantly enhanced capabilities to derive and analyze 3D data by accessing high performance computers. Initial focus is on the collection and analysis of data from a unique IVEM located at the National Center for Microscopy and Imaging Research (NCMIR). The NCMIR instrument is one of a few IVEMs in the USA made available to the biological research community by the National Center for Research Resources of the National Institutes of Health. This particular instrument was specially designed to obtain images from thick specimens containing substantial 3D structure. Because of the characteristics of the EM, images acquired represent two dimensional projections of the 3D structure within the specimen.

Recently there has been great interest in creating Internet and Intranet tools for electron microscopy. We can somewhat arbitrarily divide the goals of these tools into three categories: (1) Service lab interaction, (2) Laboratory extension, (3) Teaching & outreach. The service lab wants to disseminate the results of experiments quickly to its customers. Sometimes it needs to do this during the experiments for verification that the experiments are being correctly performed, but most collaboration problems can be solved with a phone discussion while looking at the same image. This level of interactivity is not sufficient for the laboratory extension community that wants real time experimental results and perhaps even controls of the equipment projected to a remote location. Finally, several groups have expressed interest in bringing observation and control of instrumentation into the classroom to potentially many computers and potentially many different displays of the experiment in progress.

A stop-frame animation system has been developed for producing high-resolution, color and stereo motion picture animation sequences. The first of these sequences can be seen in the recently released IMAX 3D movies, “Four Million House Guests” a.k.a. “The Hidden Dimension”. IMAX movies have long been known for their breathtaking special effects that seem incredibly realistic because of the large projection screen (about 7 stories high) which is close to the entire audience, high resolution, and powerful audio effects. IMAX 3D is an extention to the traditional format that allows the audience to see three dimensional special effects with the aid of electronically shuttered viewing glasses. IMAX movies are an ideal medium to demonstrate the high resolution digital images that are possible with the SEM.

The goal of the SEM movie project was to produce movie sequences where viewers feels like they are flying smoothly through micro-space past microscopic creatures and objects in three dimensions and in color.

In 1995, we presented the initial design and demonstrated operation of the Tele-Presence Microscopy (TPM) Collaboratory at ANL. At that time the primary interaction mode for the Collaboratory was passive operation where the user was given a fixed interaction space with the system. Since then a number of changes have been implemented in both the system and its configuration. While the TPM Site is still accessed using browser based technology (URL = http://tpm.amc.anl.gov) the decisions which have been made at ANL since then, have necessitated that restrictions to the nature of the client browser software. Due to the requirement that TPM provide both a platform independent persistent electronic space as well as real time imaging, we have chosen to implement server push technology. This means that only those browsers which are fully compliant with Netscape Version 2.0 or greater now have full access to the TPM Site (MS Internet Explorer is not compatible with this technology).

One of the primary research components of the DOE2000 Materials Microcharacterization Collaboratory involves studies of catalyst materials for reduction of NOx emissions from diesel engine exhausts. A series of experimental catalysts is currently under investigation with colleagues from Ford Research Laboratory. The major thrust of this research is the study of catalyst reactions to determine the progressive changes in the structure and morphology of the catalytic materials with temperature and time under conditions simulating diesel exhaust exposures. Because the High Temperature Materials Laboratory (HTML) provides an ex-situ reactor system that permits TEM samples to be examined after sequential reactions without intermediate atmospheric exposures, the research was undertaken on a collaboratory basis, with microscopy sessions conducted remotely via telepresence. This permits Ford scientists to interact with HTML (and/or other collaboratory) scientists during the TEM sessions, but does not require their time and involvement during the actual reactor runs. The first of these telepresence sessions was held recently, as described below.

One of the missions of the NIST Manufacturing Engineering Laboratory (MEL) is to improve and advance length metrology in aid of U.S. Industry. This responsibility is found within the Precision Engineering Division (PED). The successful development of a “Collaboratory” for TelePresence Microscopy provides an important new tool to promote technology transfer in the area of length metrology and measurement technology. NIST and Texas Instruments, under the auspices of the National Advanced Manufacturing Testbed (NAMT) and in collaboration with the University of Illinois are developing a microscopy collaboratory testbed. This facility is designed to demonstrate the value of telepresence microscopy within a large distributed manufacturing facility such as Texas Instruments and between organizations such as NIST, Texas Instruments and Universities.

Telepresence Microscopy is an application of the state-of-the-art Internet based technology to long-distance scientific endeavors.

In recent years, a number of different groups(1) have investigated and reported on implementing Web-based controls for SEMs and TEMs. Through telemicroscopy, as the field is often known, they have demonstrated that expensive microscopy resources can be shared and the experience of remote experts utilized in an efficient, cost-effective manner. Up to this point most technological demonstrations of telemicroscopy have been implemented as retrofits to existing microscopes. As retrofits, these demonstrations had limitations which a system designed from day one as a networked instruments could avoid. present here details of an ongoing project at R.J. Lee Instruments to redesign the software interface to our Personal SEM product around a TCP/IP link between the SEM hardware and the user interface. All communications between the SEM and console, including commands and image data are transmitted over a single 10-BaseT Ethernet link using the connection-oriented TCP protocol.

The West Greene School District (WGSD) in southwestern Pennsylvania has been extremely aggressive in providing exceptional educational opportunities to its students which come primarily from a rural, small-town and post-industrial mining community. The five school systems serving Greene County have been linked with multiple T-l lines making special interest courses available to all students through a sophisticated distancelearning facility. The goals of this program are multiple including motivating students to pursue advanced education, especially in mathematics, computer sciences, engineering, and the basic sciences; enriching the educational experience beyond that commonly available to students in rural communities; and improving the general attractiveness of the region for business growth.

RJ Lee Group (RJLG) is a small business located about 60 miles northeast of Greene County specializing in solving materials-related problems. Scanning electron microscopy (SEM) is often one of the methods used by RJLG.

Digital imaging has made electron microscopy more quantitative, more direct, and more automated. It has vastly changed the way in which microscopy is done today. The conventional photographic emulsion technique for image recording is being replaced by CCD-based digital imaging techniques in virtually all aspects of electron microscopy applications, resulting in increased work efficiency and data accuracy. Many of the microscope's evaluation procedures are traditionally time-consuming and complicated, such as measurements of TEM resolution and spherical aberration coefficient of the objective lens, etc. However, they can be greatly simplified with the use of slow-scan CCD cameras bee and appropriate software.

With many superior properties over other image recording media, slow-scan CCD cameras offer possibilities for many new TEM applications, such as TEM autotuning and automated microscopy. On-line image display and analysis have proved to be the key elements in CCD-based digital imaging.

Control of electron microscopes by a remote networked computer system has been implemented successfully in a variety of ways including via the World Wide Web (WWW). This approach works well with users who are familiar with the operation of the instrument and are experienced microscopists. However, many of our collaborations involve other scientists or engineers looking to us for analytical expertise. In order to accommodate collaborators with a broad range of skills and to allow them to interact remotely, in near-real time with a scientist operating an instrument, we have implemented a Virtual Collaborative Environment (VCE) based on a WWW server and the use of the server-push protocol.

The system developed at NIST is a frames-based WWW page that is presented to the user with an index of instruments on the left side (FIG.l), live video pages to the right side, and a download area (via ftp) toward the bottom for data and high resolution images.

Collaboration between local microscopists and image processing specialists, and their remote biological colleagues, has been hampered by the difficulty of i) transferring the three-dimensional reconstructions of macromolecules resulting from the cryomicroscopy and image processing, ii) viewing the results in a meaningful way, and iii) communicating the results and the interpretations derived therefrom to each other.

The acrosomal process is an intracellular quasi-crystalline organelle in the head of the sperm of the horseshoe crab Limulus polyphemus. It consists of 100 - 130 actin-scruin filaments packed together in a pseudo-hexagonal lattice and is up to 60 μm long with a diameter of 0.1 μm. Scruin-scruin interactions are responsible for cross-linking the actin filaments together in the bundle. Our goal was to reveal interfilament interactions in the bundle. We have taken tilt series images in the electron microscope to reconstruct its three-dimensional structure at 45 Å resolution.

The advent of digital imaging in microscopy has inspired many changes to the conventional generation and presentation of photographic image data. The demise of the photographic darkroom is a direct result of the prolific electronic image format. Digital images offer extensive flexibility with respect to image manipulation and analysis, duplication, technical presentations, and data transfer to remote locations. With this advance to digital technology come new obstacles for the "traditional" microscopy laboratory to hurdle once this technology is implemented. A few of these obstacles include data storage and retrieval systems, networking of analytical instruments, and adequate software for image manipulation.

The intent of this presentation is to introduce the analytical community to a successful implementation of a laboratory-wide image management system, specifically, the strategies behind the concept and completion of the system and successes gained therefrom.

We report here the successful development of a new instrument, the Centrifuge Polarizing Microscope (CPM). The CPM was developed to explore, by high-extinction polarized light microscopy, the dynamic alignment, linkage, and interaction of macromolecules, fine structure, and organelles in living cells and developing embryos being centrifugally stratified. It should also find applications in material sciences, e.g., for examining centrifugal stratification, compaction, and alignment of liquid crystals and emulsions; the CPM uniquely allows continuous observation and measurement of the weak birefringence exhibited by molecules and fine structures that are oriented, or become oriented, under centrifugal acceleration.

Centrifuge microscopes of various designs have been built since the 1930s but were not equipped for polarization optics. In the 8-cm-radius rotor of our prototype CPM, spinning between the objective and condenser lenses with numerical apertures of up to 0.55, the specimen chamber is illuminated stroboscopically by a 5- to 7-ns intense flash of 532-nm-wavelength light from a Nd- YAG laser, synchronized to fire exactly as the specimen intersects the axis of the objective lens (Fig. 1).

The collection and analysis of x-rays emitted by samples in electron microscopes is in many ways a dynamic process. Conventional methods of off-line teaching are restricted in their ability to convey this dynamic and interactive nature. The recent developments in multimedia software and the increase in power of desktop systems can be taken advantage of in teaching in this area. Specifically the original AMAS (Australian Microbeam Analysis Society) EDS course notes have been integrated with a range of interactive tutorials and multi-choice assessment using the Macromedia Director 6.0 multimedia authoring software. The ‘Virtual EDS’ software is compatible with the Macintosh, Windows'95, Windows NT and Windows 3.11 platforms, on CD-ROM.

The project is on-going but currently the software contains the full AMAS EDS notes updated as thirteen reference topics and the following four interactive tutorials:

A partnership has developed between the Microscope and Graphic Imaging Center (MAGIC) at California State University, Hayward and Ohlone Community College. The purpose of the collaboration is to develop a program to allow community college students to gain experience in preparing and viewing samples using scanning electron microscopy technology. The learning module involves students from the Ohlone College Biology Majors Program and student mentors from CSUH. An additional component is the introduction of under-represented students into a Biology Fellowship Program in which they also participate in the SEM learning module. Participants for these programs are selected on the basis of their interest and how this experience will benefit them as expressed in a one-page written essay. Ten students are selected to participate in the programs.

The objectives of the learning module are to: 1) learn specimen preparation techniques and develop skills in SEM technology; 2) gain hands-on experience and develop some laboratory skills necessary for effective use of a SEM in studying biological specimens; 3) share the experience gained with peers at their home institution.