Optical microscopy development

The screening of libraries of compounds for the desired property constitutes an essential part of the combinatorial process. The easier and the faster the screening, the higher the throughput and the more compounds can be screened in a unit of time. This paradigm has led Still s group to develop a combinatorial approach to chiral selectors that involves a visual screening step by optical microscopy that enables the manual selection of the best candidates [81]. 

Much of the difficulty in demonstrating the mechanism of breakaway in a particular case arises from the thinness of the reaction zone and its location at the metal-oxide interface. Workers must consider (a) whether the oxide is cracked or merely recrystallised (b) whether the oxide now results from direct molecular reaction, or whether a barrier layer remains (c) whether the inception of a side reaction (e.g. 2CO - COj + C)" caused failure or (d) whether a new transport process, chemical transport or volatilisation, has become possible. In developing these mechanisms both arguments and experimental technique require considerable sophistication. As a few examples one may cite the use of density and specific surface-area measurements as routine of porosimetry by a variety of methods of optical microscopy, electron microscopy and X-ray diffraction at reaction temperature of tracer, electric field and stress measurements. Excellent metallographic sectioning is taken for granted in this field of research. 

Until the advent of modem physical methods for surface studies and computer control of experiments, our knowledge of electrode processes was derived mostly from electrochemical measurements (Chapter 12). By clever use of these measurements, together with electrocapillary studies, it was possible to derive considerable information on processes in the inner Helmholtz plane. Other important tools were the use of radioactive isotopes to study adsorption processes and the derivation of mechanisms for hydrogen evolution from isotope separation factors. Early on, extensive use was made of optical microscopy and X-ray diffraction (XRD) in the study of electrocrystallization of metals. In the past 30 years enormous progress has been made in the development and application of new physical methods for study of electrode processes at the molecular and atomic level. 

Preliminary room temperature x-ray data of 0.65 Me4C00-PECH indicates that the sample presents a highly ordered smectic mesophase which was not yet completely assigned. The textures seen by polarized optical microscopy are also typical of smectic phases. Due to the very high molecular weights involved, textures specific to mesophase in thermodynamic equilibrium could not be developed within a reasonable amount of time by annealing. 

The computer age has brought about considerable innovation in the operation of laboratory instrumentation. One consequence of this is the wider acceptance and utilization of the optical microscope as a quantitative analytical instrument. A brief literature survey illustrates the diversity of disciplines and optical methods associated with the development of computer interfaced optical microscopy. This is followed by a description of how our methods of fluorescence, interferometry and stereology, nsed for characterizing polymeric foams, have incorporated computers. 

Recent developments have allowed atomic force microscopic (AFM) studies to follow the course of spherulite development and the internal lamellar structures as the spherulite evolves [206-209]. The major steps in spherulite formation were followed by AFM for poly(bisphenol) A octane ether [210,211] and more recently, as seen in the example of Figure 12 for a propylene 1-hexene copolymer [212] with 20 mol% comonomer. Accommodation of significant content of 1-hexene in the lattice allows formation and propagation of sheaf-like lamellar structure in this copolymer. The onset of sheave formation is clearly discerned in the micrographs of Figure 12 after crystallization for 10 h. Branching and development of the sheave are shown at later times. The direct observation of sheave and spherulitic formation by AFM supports the major features that have been deduced from transmission electron and optical microscopy. The fibrous internal spherulite structure could be directly observed by AFM. 

Finally, photomicrography is the culmination of the preparation of specimens for optical microscopy. This is a very technical area requiring proper illumination (41-43), focusing, choice of films, as well as exposure and appropriate film development. This critical area of microscopy should see continued technological innovations as much of photomicrography is being computerized (44,45). This effort is witnessing the concomitant... 

Seeing the surface of a catalyst, preferably in atomic detail, is the ideal of every catalytic chemist. Unfortunately, optical microscopy is of no use for achieving this, simply because the rather long wavelength of visible light (a few hundred nanometers) does not enable features smaller than about one micrometer to be detected. Electron beams offer better opportunities. Development over the past 40 years has resulted in electron microscopes which routinely achieve magnifications on the order of one million times and reveal details with a resolution of about 0.1 nm [1], The technique has become very popular in catalysis, and several reviews offer a good overview of what electron microscopy and related techniques tell us about a catalyst 12-6],... 

Transmission electron microscopy (TEM) is a powerful and mature microstructural characterization technique. The principles and applications of TEM have been described in many books [16 20]. The image formation in TEM is similar to that in optical microscopy, but the resolution of TEM is far superior to that of an optical microscope due to the enormous differences in the wavelengths of the sources used in these two microscopes. Today, most TEMs can be routinely operated at a resolution better than 0.2 nm, which provides the desired microstructural information about ultrathin layers and their interfaces in OLEDs. Electron beams can be focused to nanometer size, so nanochemical analysis of materials can be performed [21]. These unique abilities to provide structural and chemical information down to atomic-nanometer dimensions make it an indispensable technique in OLED development. However, TEM specimens need to be very thin to make them transparent to electrons. This is one of the most formidable obstacles in using TEM in this field. Current versions of OLEDs are composed of hard glass substrates, soft organic materials, and metal layers. Conventional TEM sample preparation techniques are no longer suitable for these samples [22-24], Recently, these difficulties have been overcome by using the advanced dual beam (DB) microscopy technique, which will be discussed later. 

Other more advanced microscopic techniques have been developed, including near-held scanning optical microscopy [166] and scanning probe microscopy techniques, such as atomic force microscopy and scanning tunnelling microscopy [166, 167],... 

A new optical microscopy method which enables interference contrast imaging of biological cells has been developed. The method is based on a well known physical phenomenon, white light interference on a thin transparent film. 

Powerful methods that have been developed more recently, and are currently used to observe surface micro topographs of crystal faces, include scanning tunnel microscopy (STM), atomic force microscopy (AFM), and phase shifting microscopy (PSM). Both STM and AFM use microscopes that (i) are able to detect and measure the differences in levels of nanometer order (ii) can increase two-dimensional magnification, and (iii) will increase the detection of the horizontal limit beyond that achievable with phase contrast or differential interference contrast microscopy. The presence of two-dimensional nuclei on terraced surfaces between steps, which were not observable under optical microscopes, has been successfully detected by these methods [8], [9]. In situ observation of the movement of steps of nanometer order in height is also made possible by these techniques. However, it is possible to observe step movement in situ, and to measure the surface driving force using optical microscopy. The latter measurement is not possible by STM and AFM. 

The main objective of the present section, however, is to begin with a very standard technique such as optical microscopy and to use it to illustrate why colloids are difficult to see and what modern developments have emerged in recent years to allow us to see and do things that were considered impossible until a decade ago. We also use this opportunity to review briefly some new techniques that are currently available to measure interaction forces between particles directly. We appeal to some of these techniques in other chapters when we discuss colloidal forces. 

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