Staining technique

Fig. 26. Photomicrograph of a series of cross sections of beefwood (Casuarina equisetifoUa) leaves, showing results with a variety of staining techniques.

Electron and optical microscopes are being used to see blend homogeneity. Elastomer-plastic blends are somewhat easier to identify than elastomer-elastomer blends because normal staining techniques, e.g., osmium tet-raoxide, can be used in the case of plastic-elastomer blends. Normally, there are two methods that are followed for examining the blend surface by electron microscopy. 

The presence of actual clumps of mold rot in a comminuted product can be demonstrated by use of the rot fragment count, which employs a staining technique and observation at about 50 X magnification. 

Chen S., Cao T., and Jin Y., Ruthenium tetraoxide staining technique for transmission electron microscopy of segmented block copoly(ether-ester), Polym. Commun., 28, 314, 1987. 

A combination of specialized staining techniques and high-resolution microscopy has allowed geneticists... 

Acid-fast bacillus General term for mycobacteria based on staining technique. 

The second step in 2D electrophoresis is to separate proteins based on molecular weight using SDS-PAGE. Individual proteins are then visualized by Coomassie or silver staining techniques or by autoradiography. Because 2D gel electrophoresis separate proteins based on independent physical characteristics, it is a powerful means to resolve complex mixtures proteins (Fig. 2.1). Modem large-gel formats are reproducible and are the most common method for protein separation in proteomic studies. 

Figure 2.1. Schematic illustration oftwo-dimensional gel electrophoresis. Proteins are extracted from the organism of interest and solubilized. The first dimension separates proteins based on isoelectric point. The pi strip is reduced and alkylated and applied to an SDS-PAGE gel for separation by molecular weight. Proteins canbe visualized using a number of staining techniques.

Another limitation of 2D gels is that membrane proteins are underrepresented. Because membrane proteins account for approximately 30% of total proteins (Wallin and Von Heijne, 1998), this is a serious problem for characterization of the proteome. The relative lack of membrane proteins resolvable on 2D gels can be attributed to thee main factors (i) they are not abundant, and therefore are difficult to detect by standard staining techniques, (ii) they often possess alkaline pi values, which make them difficult to resolve on the pH gradients most often used for isolelectric focusing, and (iii) the most important reason for under representation may be that membrane proteins are poorly soluble in the aqueous media used for isoelectric focusing (Santoni et al., 2000). Membrane proteins are designed to be soluble in lipid bilayers and are therefore difficult to solubilize in water-based solutions. 

Tlie morphology of some bacteria, especially those that form spores, is distinctive enough under the light microscope to have value for identification. This means that differential staining techniques, such as the Gram stain or acid-fast stain, and fluorescence microscopy may help to determine the iden-... 

Holm, C. Jespersen, L. A flow-cytometric gram-staining technique for milk-associated bacteria. Appl. Environ. Microbiol. 2003, 69, 2857-2863. 

Much of the confusion was caused by the inadequacy of the staining procedures. By means of the HCl-Giemsa staining technique of Piekarski, many observations have been reported demonstrating the presence of chromatinic structures in bacteria. 

Table 2.1. Filarial nematodes infected with intracellular bacteria. The method of detection is shown. Neither electron microscopy nor the immuno-histochemical staining techniques used are to be regarded as Wolbachia specific (see note). Positive identification of intracellular bacteria as Wolbachia is shown only where PCR amplified products of rRNA or ftsZ genes have been sequenced. ...

During the past two decades, cell-biological and biomedical research has greatly benefited from innovations in fluorescence microscopy. Both the increase in the repertoire of fluorescence staining techniques at the (sub)cellular level and the development of a multitude of novel fluorescence microscopy techniques contributed significantly. 

TRITC has been used in numerous applications involving fluorescence detection, including double-staining techniques with fluorescein-labeled probes (Mossberg and Ericsson, 1990), the synthesis of fluorescently labeled DNA probes (Smith et al., 1985), as a label in homogeneous... 

The intense Texas Red fluorophore has a QY that is inherently higher than the tetrameth-ylrhodamine or Lissamine rhodamine B derivatives. Texas Red s luminescence is shifted maximally into the red region of the spectrum, and its emission peak only minimally overlaps with that of fluorescein. This makes Texas Red derivatives among the best choices of labels for use in double-staining techniques. 

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