We are using the term "Industrial Polymers" to refer to polymers [plastics] that are produced by the ton or (in the case of films) by the mile. For example, in descending order of world-wide use (tonnage), the top eight of these polymers are polyethylene (PE), polyvinyl chloride (PVC), polypropylene (PP), styrene polymers (including polystyrene - PS, and acrylonitrile-butadienestyrene/ styrene-acrylonitrile - ABS/SAN), polyesters (PETP), polyurethane (PU), phenolics and aminoplastics.

Industrial polymers, which have been produced by the millions of tons for the last five decades and are of obvious social and economic importance, have been exhaustively characterized. Structural features which affect physical properties and indicate process variables have been studied by many techniques other than microscopy (x-ray diffraction, thermal analysis, rheology, chromatographies, etc.). Microscopy techniques for polymer characterization have been well documented. Our motivation to apply field emission (high resolution) scanning electron microscopy to the study of polymers is: (1) The application of low voltage, high resolution SEM to biological materials is well characterized.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

The Chlamydomonas zygospore is formed after gametic cell fusion by a progression of ultrastructural and physiological changes, including the synthesis of a protective and elaborately sculptured secondary wall (Figure 1). Assembly of the secondary wall occurs within the confines of a transient primary zygote wall, synthesized immediately after gamete fusion, and assembled beneath the residual gamete walls. Upon completion of secondary wall assembly, the primary zygote wall is shed from the maturing zygospore (Figure 2). This primary zygote wall, composed primarily of a β-1,3 glucan (callose), may be important in the regulation of secondary zygospore wall formation.

A temperature-sensitive mutant strain of C. monoica, zym30, has been isolated that shows marked abnormalities in both primary and secondary wall ultrastructure. The degree of wall formation in the mutant strain is inversely proportional to temperature. At 25° C most zygotes fail to synthesize either the primary or the secondary wall. At 15° C both primary and secondary wall assembly occurs to some extent and as many as 50% of the zygotes develop a “pseudo-wildtype” phenotype (Figure 3). Although release of the primary wall rarely occurs in the zym30 strain (even at permissive temperatures), those walls that are released appear thinner than the walls shed from wildtype zygospores, as viewed by phase contrast microscopy. Fluorescence images incorporating aniline blue, a fluorochrome specific for β-1,3 glucans, show decreased fluorescence suggesting reduced callose synthesis (Figure 4).

Several permanent, ultrastructurally distinct layers comprise the secondary wall, the assembly of which may be controlled, in part, by the primary wall. One or more of these secondary wall layers contain the highly durable, autofluorescent biopolymer, sporopollenin. Autofluorescence of zym30 zygospores produced at the permissive temperature is enhanced relative to wildtype zygospores. This correlates with ultrastructural changes in the primary wall and overproduction and/or misassembly of the surface secondary wall layer (Figure 5). Work is in progress to determine whether the environmental resistance, and/or viability of the zym30 “pseudowildtype” zygospores are reduced relative to wildtype zygospores.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Most endosymbionts remain separated from their host cell by a derivative of the phagocytic membrane. This membrane and its contents have been defined as a novel “organelle-like” structure called a symbiosome. However, in the case of many cnidarian-dinoflagellate symbioses the putative symbiosome is composed of multiple layers of apparent membranes (Fig. 1). In the past the origin of these membranes has been assigned to host and/or symbiont with very little experimental evidence to support either view; however, several interesting hypotheses have been proposed to explain the origin of the “membranes”. Trench and Blank suggest that occasionally the outer most layer of the vegetative symbiont's cell wall is sloughed off the surface of the cell. They concede that the outer most layer of “membranes” must be the host vacuolar membrane, but believe that it simply prevents the sloughed layer from moving away from the cell, (as would happen in culture), and that, over time, multiple sloughed layers accumulate beneath the host vacular membrane.

Cells of Pseudomonas aeruginosa were exposed to six different heavy metals, Al, Cd, Cu, Mn, Pb, Zn, exposure was done at the same concentrations of 20 ppm, except for the Cd which was done at 5 ppm. The cells were then analyzed using the STEM mode of a transmission electron microscope in conjunction with a PGT IMIX energy dispersive xray spectrometer. The data was then analyzed using a bulk sample analysis program (ZAF method) in standardless mode (w/w). We analyzed the cell wall the cytoplasm and polyphosphate bodies. Approximately 20 cells were analyzed per treatment and averages of the elements present were calculated. The cells of Pseudomonas aeruginosa were grown in nutrient broth and then embedded in Epon according to Luft's procedure. Quantitative analysis of the three different parts of the cell revealed that in cells exposed to Al, the wall contained on average 0(55%), C(20%) P( 8.0%) K(10%) Cl(2%) S(5.0%),Al(2.0%), the cytoplasm contained on average, 0(53%) C(20%) P(5.0%) Ca(4.0%) K(12%) Cl(4.0%) Al(2.0%).

Digital microscopy, the integration of digital and microscopy technologies, was initiated for quantitative microscopic image analysis, but it is now for almost all microscopy applications. During the past decade, with the advance of digital technologies, digital microscopy imaging is becoming an indispensable technology in drug discovery.

We started establishing state-of-the-art digital microscopy imaging for drug discovery with the investigation of bioimaging applications at our US research site. Our results shown that all the top 5 bioimaging needs require computer-aided microscopy. Based on this investigation and our review of the microscopy imaging applications in the pharmaceutical industry, we determined four directions for microscopy in drug discovery: multidimensional/multimodal microscopy, digitalization, automation, and bioimage informatics.

Multidimensional/multimodal microscopy imaging is required by the nature of biological research, which is fundamental in drug discovery. From genomic imaging to pathology observation, we require biological details and compound activities at the levels from subcellular organelles to organ tissues, from cellular signaling to anatomical locations of compounds.