Development microscopy

Virchow, who was largely responsible for the acceptance of the cell theory, developed microscopy of cells from normal and diseased tissues as a major tool (histopathology) in the clinical armory. He believed the vital functions of the cell, growth, maintenance, and multiplication were discharged by its nucleus the specialised, distinguishing functions were made possible by the extranuclear constituents. In a Sunday evening lecture in Edinburgh in 1868, On the Physical Basis of Life , Thomas Huxley described cells as protoplasmic masses usually... 

The practical applications of the various microscopical techniques have created opportunities for microscopists in industry and, in particular, within pharmaceutical research and development. Microscopy is used extensively, from the earliest stages of drug discovery into late development and even into manufacturing. Pharmaceutical microscopy can be conveniently divided into physico-chemical and biological applications. This chapter will consider exclusively the physico-chemical aspects of microscopy in the pharmaceutical industry. There are three broad areas in which microscopy can play an important role in the development of drugs solid-state analysis, particle size and morphology studies, and contaminant identification. This chapter presents an overview of how microscopy contributes to each of these three areas. The emphasis will be on practical examples taken from the literature and from the author s experience. 

The objective of late stage solid-state analysis is to provide data for the choice of form for development. Microscopy has much to contribute to this analysis but generally plays an adjunct role to other techniques. Microscopy can, however, play a crucial role in some of the thermodynamic investigations leading... 

Microscopy may be the most versatile and most widely used method of characterizing sintering. However, because of the broad range of chemical and physical changes that occur during densification and microstructure development, microscopy is often supplemented with a variety of other characterization techniques. 

The development of scanning probe microscopies and x-ray reflectivity (see Chapter VIII) has allowed molecular-level characterization of the structure of the electrode surface after electrochemical reactions [145]. In particular, the important role of adsorbates in determining the state of an electrode surface is illustrated by scanning tunneling microscopic (STM) images of gold (III) surfaces in the presence and absence of chloride ions [153]. Electrodeposition of one metal on another can also be measured via x-ray diffraction [154]. 

Measuring the electron emission intensity from a particular atom as a function of V provides the work function for that atom its change in the presence of an adsorbate can also be measured. For example, the work function for the (100) plane of tungsten decreases from 4.71 to 4.21 V on adsorption of nitrogen. For more details, see Refs. 66 and 67 and Chapter XVII. Information about the surface tensions of various crystal planes can also be obtained by observing the development of facets in field ion microscopy [68]. 

The following several sections deal with various theories or models for adsorption. It turns out that not only is the adsorption isotherm the most convenient form in which to obtain and plot experimental data, but it is also the form in which theoretical treatments are most easily developed. One of the first demands of a theory for adsorption then, is that it give an experimentally correct adsorption isotherm. Later, it is shown that this test is insufficient and that a more sensitive test of the various models requires a consideration of how the energy and entropy of adsorption vary with the amount adsorbed. Nowadays, a further expectation is that the model not violate the molecular picture revealed by surface diffraction, microscopy, and spectroscopy data, see Chapter VIII and Section XVIII-2 Steele [8] discusses this picture with particular reference to physical adsorption. 

Rather sophisticated techniques have developed for both microscopies, too involved to discuss here. E)etails may be found in monographs such as Ref. 25, and references cited in Section VlII-2. 

One of the more recent advances in XPS is the development of photoelectron microscopy [ ]. By either focusing the incident x-ray beam, or by using electrostatic lenses to image a small spot on the sample, spatially-resolved XPS has become feasible. The limits to the spatial resolution are currently of the order of 1 pm, but are expected to improve. This teclmique has many teclmological applications. For example, the chemical makeup of micromechanical and microelectronic devices can be monitored on the scale of the device dimensions. 

AFM measures the spatial distribution of the forces between an ultrafme tip and the sample. This distribution of these forces is also highly correlated with the atomic structure. STM is able to image many semiconductor and metal surfaces with atomic resolution. AFM is necessary for insulating materials, however, as electron conduction is required for STM in order to achieve tiumelling. Note that there are many modes of operation for these instruments, and many variations in use. In addition, there are other types of scaiming probe microscopies under development. 

Raman microscopy is more developed than its IR counterpart. There are several reasons for this. First, the diffraction limit for focusing a visible beam is about 10 times smaller than an IR beam. Second, Raman spectroscopy can be done in a backscattering geometry, whereas IR is best done in transmission. A microscope is most easily adapted to a backscattermg geometry, but it is possible to do it in transmission. 

Turrell G and Corset J (eds) 1996 Raman Microscopy Developments and Applications (New York Academic)... 

The history of EM (for an overview see table Bl.17,1) can be interpreted as the development of two concepts the electron beam either illuminates a large area of tire sample ( flood-beam illumination , as in the typical transmission electron microscope (TEM) imaging using a spread-out beam) or just one point, i.e. focused to the smallest spot possible, which is then scaimed across the sample (scaiming transmission electron microscopy (STEM) or scaiming electron microscopy (SEM)). In both situations the electron beam is considered as a matter wave interacting with the sample and microscopy simply studies the interaction of the scattered electrons. 

It is interesting to note the analogy of developments in light microscopy during the last few decades. The confocal microscope as a scaiming beam microscope exceeds by far the nomial fluorescence light microscope in resolution and detection level. Very recent advances in evanescent wave and interference microscopy seem to promise to provide even higher resolution (B1.18). 

Guokenberger R, Flartmann T and Knapp FI F 1995 Recent developments Scanning Tunneiiing Microscopy ii ed R Wiesendanger and Fl-J Guntherodt (Berlin Springer) oh 9... 

A completely new method of determining siufaces arises from the enormous developments in electron microscopy. In contrast to the above-mentioned methods where the surfaces were calculated, molecular surfaces can be determined experimentally through new technologies such as electron cryomicroscopy [188]. Here, the molecular surface is limited by the resolution of the experimental instruments. Current methods can reach resolutions down to about 10 A, which allows the visualization of protein structures and secondary structure elements [189]. The advantage of this method is that it can be apphed to derive molecular structures of maaomolecules in the native state. 

The slit-shaped model has come into prominence in recent years, as electron microscopy has revealed the prevalence of solids composed of platelike particles the technique, indeed, has now developed to the point where it is possible to identify the presence of slit-shaped pores, and even to measure their width. In the ideal case where the sides of the slit are truly planar and parallel, the hysteresis takes an extreme form since the mean radius of curva-... 

Nobel-laureate Richard Feynman once said that the principles of physics do not preclude the possibility of maneuvering things atom by atom (260). Recent developments in the fields of physics, chemistry, and biology (briefly described in the previous sections) bear those words out. The invention and development of scanning probe microscopy has enabled the isolation and manipulation of individual atoms and molecules. Research in protein and nucleic acid stmcture have given rise to powerful tools in the estabUshment of rational synthetic protocols for the production of new medicinal dmgs, sensing elements, catalysts, and electronic materials. 

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