The Environmental Scanning Electron Microscope (ESEM) is capable of image generation in a gaseous environment at sample chamber pressures of up to 20 torr. in an ESEM, low energy secondary electrons emitted from a sample surface, by virtue of the primary electron beam, are accelerated towards the positively biased metallic ring (typically +30 to +550V) Gaseous Secondary Electron Detector (GSED). As these electrons accelerate towards the ring they undergo ionizing collisions with gas molecules producing positive ions and additional electrons known as environmental secondary electrons. The environmental electrons further ionize the gas on their way to the ring producing a cascade amplification of the original signal. The amplified signal induced in the ring is used to form an image. The electric field generated between the GSED ring and the grounded stage causes the positive ions produced in the cascade to drift towards the sample, effectively neutralizing negative charge build up on the surface of a non-conducting sample.

An increasing body of experimental evidence indicates that the flux and distribution of gaseous ions significantly affects secondary electron contrast of dielectric materials imaged in low vacuum and environmental scanning electron microscopes. Unusual contrast has been identified in some insulating materials, and attributed to charging effects (1-3). However, as these contrast mechanisms have not been reported in conventional microscopes, it would seem that the ions play an important role in making the contrast manifest. The effects of ions can include the creation of space charge in the gas above the specimen, influencing the charge state of the specimen surface, and electron-ion recombination (possibly leading to energy filtering of emitted secondary electrons). Space charge and surface potential can influence the gain realised in the gas amplification cascade by altering the detector field (4). However, surface potential, and recombination affect the number of electrons injected into the amplification cascade, to give rise to contrast variations in the final image.

The possibility of performing scanning electron microscopy under “environmental” conditions (ESEM); where the specimen does not have to be under high-vacuum conditions, or even coated to make it conductive, has existed for several years [1]. in particular it has been possible to image in conditions that allow the samples to remain fully hydrated during observation. This unique feature of the ESEM has been exploited to image the dehydration of aqueous protein gel materials which have been created under controlled conditions to produce a range of Gel types.

Drops of several μl of the Gels were placed in the Peltier cooling stage of the FEI-Philips Electroscan E3, and allowed to stabilize at 4.1°C. Several drops of water were placed on the stagesurround to maintain a high vapor pressure of water around the sample. The chamber was then pumped down to the working pressure of 6 torr and then back-filled with pure water vapor following the technique of Cameron et.al. [2].

We present experimental evidence for the effects of electron-ion recombination on contrast formation in secondary electron (SE) images obtained using an environmental scanning electron microscope (ESEM). Inclusion of the effects of SE-ion recombination in the theory of ESEM image formation accounts for contrast reversal observed, under appropriate conditions, in SE images of conductors (ie, in the absence of localized specimen charging) and of insulators that exhibit localized charging. Previously unexplained dynamic contrast caused by temporal charging can be understood if both conventional models of the effects of charging on SE emission, and the proposed role of SEion recombination in ESEM image formation are accounted for.

In an ESEM, the extent of charging exhibited by insulating specimens can be controlled by varying the type and pressure (P) of the imaging gas, and operating parameters that determine the extent to which the gas is ionized (eg, detector bias, VGSED, and working distance, WD).

In a focused ion beam (FIB) instrument, ions (typically Ga+) obtained from a liquid metal ion source are accelerated down a column at energies up to ∽ 50 keV. The beam of ions is focused by electrostatic and octopole lens systems and the ion dose (and beam diameter) is controlled using real and/or virtual apertures. Beam sizes in FIB instruments on the order of 5-7 nm may be achieved.

The versatility of the FIB instrument enables large regions of material (e.g., 500 μm3) to be removed at high beam currents in just a couple of minutes. Lower beam currents (i.e., beam diameters) are usually used to remove smaller amounts of material within the same time frame (e.g., ∽ 5μm3). The introduction of an organometallic gas in close proximity to the target allows for the deposition of metals, SiO2, and other materials, by an ion beam assisted chemical vapor deposition process.

A microscope facility at teaching institutions needs to be more than an isolated refuge of researchers. Students must be given easier access to the equipment and an increased awareness of the power of these tools for understanding the scientific principles taught in their lecture classes. The San Diego State College of Sciences supports a shared instrumentation Electron Microscope Facility, a suite of seven rooms that occupies 1500 square feet. The laboratory is fully equipped for all aspects of sample preparation for examination by confocal scanning light microscopy, scanning electron microscopy (SEM) or transmission electron microscopy (TEM). Historically, specialists with many years training in equipment operation, sample preparation, and research protocols have had exclusive access to EM facilities. At SDSU, however, every effort is made to encourage inclusivity via familiarity with and hands-on access to equipment in the EM Facility. in all, over 1000 SDSU students and several hundred community members participate each year in opportunities to access the SDSU EM Facility.

The Biosciences Electron Microscopy Facility at UBC has at present two TEMs, one SEM, two confocals. several fluorescent light microscopes and an above average amount of digital imaging equipment used p by graduate students. The facility supports the Faculty of Medicine and the Faculty of Science and is run by a Director (myself) with one assistant. It is busy having an average of 600 individual users per year. The only way this number of users can be accommodated is by making them independent of staff. Training students had been previously on a one-to-one basis. This is very time consuming. Four years ago with no full time assistant, I decided to approach training in a different way. We started a confocal microscopy workshop aided by one PhD student.

We run a 1.5 day practical confocal workshop. It is one day as a group, giving the basic concept of confocal microscopy, and using the instrument from “this is how you switch the machine on” to “this is how you deal with the stack of images.”

For many years the means of communicating electron microscopy techniques had been through either the spoken or the written word. with the advent of the computer and in particular the notebook computer more exotic methods of information presentation were made available. Now with the compact disk or the DVD, with their ability to store very large quantities of data, even more new avenues have been opened up for trainers.

There are a number of programs by small organisations, like QM Slide Show and EZ Slide, that enable the trainer to take the simple approach to presentation preparation. Using a Windows Notepad interface or similar the trainer gathers bitmap files in a simple sequence, referencing the file source to activate each part of the presentation. This type of presentation is extremely flexible, so much so that the trainer, with a large selection of bitmap files and a good filing system, may develop a completely new or modified presentation in just a few minutes.

Two of the greatest challenges facing any educator are deciding what information will be included in a syllabus and how to present it for maximum learning. Distilling the experience derived from a quarter of a century teaching microscopy exposes one driving concept: the importance of education, not just training.

The difference between these two approaches is fundamental. Education involves laying a foundation on good science, building a vocabulary for better communication and future growth, and engendering a questioning attitude about the probe-matter interactions which, in turn, leads to valid interpretation of the information in an image. Training is a matter of learning what knob to turn, which button to push: an approach which overlooks the sample itself; how the light, electron, or scanning force is interacting with that sample; and what can be learned as a result.

In a two year microscopy teaching program which includes 18 microscopy courses, there are thousands of images which must be collected from a variety of input devices running on various platforms. These must then be associated with different types of data files that need to be tagged to reports as well as being tagged to projects and courses. All of this information must be accessible, first to all those involved with doing the project as well as to anyone associated with needing the results. Tracking progress and security issues had to be part of the solution. It was also necessary that all of this take minimum staff time. Thus, we had to not only set up an image database, but truly an asset management system. Although it certainly would be possible to develop one’s own system, we are limited in staff time for such a project, and certainly didn’t want to re-invent the wheel if such a product was commercially available.

The West Greene School District (WGSD) in partnership with RJ Lee Group (RJLG) is developing innovative programs as part of the Schoolhouse Project to provide exceptional educational and business opportunities to its students. The objectives of the educational/business partnership 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. Access to remote analytical instrumentation is a key component of the Schoolhouse Project. to this end, RJLG has developed Web-based technologies (WEBSEM™) which enable control and operation of a scanning electron microscope (Personal SEM® or PSEM) over the Internet. A key component of the project will be the establishment of an incubator laboratory extension of RJLG at the WGSD. This laboratory will function as a learning laboratory during the school day and as a business during the evenings and weekends.

Abstract: Tele-microscopy has become a very active area of research and development in the 1990s where there was a desire to give researchers better access to expensive and specialized microscopes, breaking geographical and time barriers. Rapid advances in telecommunication, computers, and microscopy technologies make possible the establishment of the World Wide Web and the realization of the goal of developing a “laboratory without walls”. This provides unprecedented opportunities for researchers and educators alike to gain access to shared instrumental and educational resources.

In this presentation, a description of the Interactive Nano-Visualization for Science and Engineering Education (IN-VSEE) project will be given. The primary goals of IN-VSEE are to (i) convey the excitement of nanoscience and nanotechnology to promote studentmotivated learning and pursuit of science and engineering careers, (ii) teach fundamental interdisciplinary concepts in science and engineering using a visual format to help students learn and integrate information more effectively, (iii) provide students with the capability to routinely explore materials in three dimensions with resolutions at the nanoscale and even down to the atomic scale, and (iv) demonstrate the feasibility of remote operation of research-grade laboratory instrumentation for development into a powerful educational and collaborative tool.

The W.M. Keck Foundation Center for Molecular Structure (CMolS) at California State University Fullerton is the first comprehensive x-ray crystallographic and computational facility located at a predominantly undergraduate institution. CMolS, a core facility of the California State University Program for Research and Education in Biotechnology (CSUPERB), engages more than thirty faculty and hundreds of student investigators throughout the 23-campus CSU in joint research and education activities directed at the determination and critical analysis of molecular structures. Recognizing that distance learning and remote access represent an exciting new area of research and training, CMolS has taken a leadership role in the CSU in using the Internet to restructure and globalize formal education in x-ray crystallography by developing and implementing novel Webbased courses, tutorials and virtual laboratories that require specialized instrumentation and expertise.

An NIH Structural Biology report and the Department of Energy [1] have indicated that high throughput crystallography is a top research priority in the post-genomic era.

The University of Iowa Central Microscopy Research Facility (CMRF) has been in existence for 27 years. Starting out as a transmission electron microscopy (TEM) research laboratory, the facility has offered formal college courses. These courses require that students identify a project to investigate during the semester. Theory from the formal lecture is reinforced by work performed in the laboratory session. From its modest beginnings, the CMRF has continually grown. Currently, the facility offers two Confocal microscopes, two Scanning Electron Microscopes, a Scanning Probe Microscope, Energy Dispersive Spectroscopy, a Mossbauer spectrometer, a PTI Ion Imaging/Ratio system, a Freeze Fracture apparatus, and three light microscopes equipped with CCD cameras. Techniques range from routine histology to in-situ hybridization. Technological advances over the history of the facility have not been confined to the lab. in the past, most lectures were given using overheads and 35mm slides.

For the past two years we have been operating a remote instrument educational project called Bugscope. Bugscope is an educational outreach project that provides access to an environmental scanning electron microscope (ESEM) for K-12 classrooms. While the operational aspects of the project require a minimal amount of staff time, the information management for the project is difficult for a small microscopy research group to support without a significant allocation of resources away from the group’s principal research goals. in an effort to alleviate this problem we have begun, in the past five months, to develop a software toolkit called ‘Information Technology for Outreach Projects’ (ITOP) - using the Bugscope project as a test bed. The goal of ITOP is to make it practical for academic research groups to provide scientific resources for educational outreach projects by automating many of the administrative and data handling tasks.

ScienceFest is coordinated through the International JASON Project that introduces the world of science through live satellite broadcasts. Dr. Robert Ballard, the ocean explorer who found the Titanic, started the program in 1989. Included in the ScienceFest presentations at the John Ford Bell Museum of Natural History, Project MICRO run by the Minnesota Microscopy Society (MMS) was on display to the public. As in past years [1] the Project MICRO team of the MMS set up various microscope stations and invited the participants to record the things that they saw and to note what they learnt and mention the things they would like to see. The microscope stations are quite simple, low power ‘field’ microscopes and various samples as documented by the “Microscopic Explorations” [2] handbook that has been developed by the MSA in collaboration with the Lawrence Hall of Science [3] as part of their Great Explorations in Math and Science (GEMS) series.

Three rigorous one-semester, graduate-level courses are offered in light microscopy (LM) and scanning (SEM) and transmission electron microscopy (TEM), primarily for graduate students and staff, and for advanced undergraduate students in the teaching complex of the Bessey Microscopy Facility (BMF). These courses have evolved over the years. They include a variety of techniques and laboratory exercises that prepare a successful student to become an independent researcher in the BMF or elsewhere, and to work with any major professor who may not have expertise in microscopy.

Each course consists of: five credits at the 600 level; a limit of 10 students; 11 hours of formal lecture-lab time per week; and additional arranged time for continuation of laboratory procedures and individual operation of microscopes and ancillary equipment. Each student has access to the teaching complex 24 hours a day, seven days a week.

This invited paper will address several emerging technologies that have an impact on scientists moving into digital imaging technology and raise a few concerns about this digital technology that may not have reached our group.

The quality of inkjet printers has moved forward at a staggering rate. The latest printers from Epson (the 900 series) have achieved a milestone in image quality that is very significant and yet is difficult to find in the specification sheet. The “dots” which the company claims can be as small as 3 picoliters are sufficiently small that they cannot be seen with the naked eye. with an improved algorithm, the image quality in 1440 × 720 mode is as good as any dye-sublimation printer. The current 980 when printing in 1440 mode prints at twice the speed as the older 900. At only $250, it actually becomes cost effective to replace 900's with 980's solely based on the amount of time that is saved (even at a University).

Low cost and reliability, optical storage media, in particular the CD-R, has become the main steam data storage media. This presentation is intended to discuss issues related to the CD-rewriteable technology and the longevity of the data stored.

The working principle of CDs can be subdivided into three major categories, the CD-ROM, CD-R and CD-RW. The CD-ROM is primary used for music and software distribution and is manufactured by injection molding process. in contrast, CD-R and CD-RW are storage media allows data storage by users. The manufacture process involves spin-coating a layer of photo-sensitive organic dye on a polycarbonate substrate, then over-coated with a refractive coating. Writing of the data bits was achieved by photo-bleaching processes, which change the absorption properties of the dye. Considering the operation principle, there are many factors influence the reliability and storage life of the CD-R.

Direct-imaging cryo-electron microscopy combined with CCD cameras provide a powerful tool for studying macromolecular assemblies in the 2 to 300 nm range, which form in biological systems, biomedical solutions, surfactants, microemulsions and a host of other diverse systems. The CCD cameras are especially advantageous for identifying rare assemblies, studying specific assembly details (figures A and B) and transient structures forming during dynamic processes and phase transformations (figures C-E). This setup is also beneficial for exploring coexistence of structures and studying highly radiation-sensitive systems such as oil-continuous solutions. Information on internal ordering is also readily obtained (figure A).

Continuous CCD application at low dose conditions provides numerous significant advantages over classical imaging on photographic plates. First, it allows a preview of the exact area to be recorded (search and focus modes), concentration on specific interesting features and adjustment of microscope parameters for optimal imaging (focus mode), at very low exposures and with minimal radiation damage.