Stain patterns

Takami S Graziadei P. and Ichikawa M. (1992). The differential staining patterns of two lectins in the accessory olfactory bulb of the rat. Brain Res 598, 337-342. 

Figure 4 shows confocal images of the staining pattern for DDC (Fig. 4A), serotonin (Fig. 4B), and TH (Fig. 4C) in the segmental ventral ganglion of the CNS from third instar larvae. Panels B and C are the same CNS double stained with serotonin and TH. The DDC-expressing cells can be categorized into a set of paired ventral lateral serotonin cells (Fig. 4A,B labeled VL in 4A), and two morphologically distinct types of dopamine cells, the medial dopamine cells (Fig. 4A,C labeled M) and the dorsal-lateral dopamine cells (Fig. 4A,C labeled DL). Figure 4 demonstrates clearly that individual DDC cells synthesize either serotonin or dopamine, but not both. One additional set of cells shows TH immunoreactivity in the ventral ganglion. These six large vacuolated cells are located more laterally than any other DDC cells (Fig. 4C, unlabeled short arrows). It is likely that the immunoreactivity in these cells results from a non-specific cross-reaction, since these cells are not...

Mailer Why do you say that the first spindle poles are made of PCM Is there some reason to think that PCM is needed without a centrosome In Nenopus, I have never heard anyone argue that there is PCM organizing the meiotic spindles. If you look at the /-tubulin staining pattern, it is totally different in a meiotic spindle than in a mitotic spindle. 

Table 5.1 lists a number of SG and PB markers, and indicates the relative specificity of each reagent for SGs or PBs. This is only a partial list of known SG and PB markers, focusing on some that are broadly useful and for which antibodies are readily available. The staining patterns obtained using some of these antibodies are shown in Fig. 5.1. 

At Leica Biosystems Newcastle Ltd., invasive breast cancer tissue controls, demonstrating HER2 expression levels at 3+, 2+, 1+, and 0, are incorporated into all Oracle HER2 Bond IHC System cell line quality control runs. This ensures that control cell lines are validated as a viable assay control. The evaluation of control cell lines should always be performed within the context of appropriate tolerance limits. Subtle changes from batch to batch may occur, and it is the correct evaluation of the cell line staining patterns within appropriate tolerance limits that enables control cell lines to be utilized both in a commercial setting and as an EQA monitoring device. 

Figure 6.10 MDA-MB-175 (1+) cell line demonstrating unique glandular-like luminal formation with correct HER2 IHC staining pattern highlighted. Stained with Dako HercepTest K5204. Blue arrows indicate specific weak incomplete 1+ membrane staining, whereas the green arrows illustrate nonspecific moderate luminal surface staining, which is not interpreted. See color insert.

Optional when using a fluorescent label, a short treatment (1 3 min) with 4% formaldehyde in PBS before mounting in water-soluble media is recommended for blocking the detachment of the fluorophore from the antibody this preserves the staining pattern for a longer storage. Wash sections in PBS for 2x3 min. 

Staining patterns of two-dimensional gels show the six RC ATPases to be present at comparable levels, and affinity capture of yeast 26S proteasomes indicate the presence of one copy of each in the regulatory complex. Mutational analysis in... 

Close observations of immunofluorescence signals showed that there are three different staining patterns, which correspond to three different nucleolar compartments FC (Fibrillar center), DFC (dense fibrillar component), and GC (granular component). Nucleolus is surrounded by heterochromatin. When the cells are in very active state of its proliferation, the nucleolar compartments and heterochromatin are integrated into a highly intricate structure called nucleolonema . A recent study has suggested that the chromatin associated with the nucleolus is less mobile than... 

In addition to the many enzyme systems available, there are with each a series of chromogenic substrate solutions that can be used to create different colors and locations of reaction products. For the peroxidase system, there are numerous oxidizable compounds that precipitate as a permanent color. The most common and still widely used is 3,3 diaminobenzidine tetrahydro-chloride (DAB). This compound precipitates to a golden brown color when in solution with peroxidase and hydrogen peroxide. This brown color has many subtleties and readily stands out in a tissue section. With practice, it is possible to differentiate specific from nonspecific staining patterns just by examining the characteristics of the precipitated pigment. This material is also insoluble in alcohol and xylene, and therefore the tissue may be routinely dehydrated and cleared without loss of chromogen. 

Microwave heating Nonspecific granular cytoplasmic staining pattern is often accentuated when paraffin sections are microwave-pretreated (5). A biotin block... 

PCR and in vitro recombination reactions are quite simple and straightforward for generating multiple expression plasmids in parallel, e.g., in a 96-well plate see Fig. 3a). The first preliminary expression experiment was done to evaluate the production level of each GST-fused protein. In this step, we compared the staining patterns of E. coli proteins harboring expression plasmids with the patterns of proteins harboring empty vectors on sodium dodecyle sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under the same culture conditions see Fig. 3b). In addition to... 

Figure 26-15 (A) Numbering and staining patterns (G-banding) of human chromosomes. The horizontal line marks the centromeres. Chromosomes 13,14,15, 21, and 22 have nucleolar organizers located at the constrictions in their short arms. From Alberts et al.297 Adapted from U. Franke.298 (B) Map of genetic defects identified in X chromosomes by 1995. From McKusick and Amberger.294...

Accessibility and preservation of the nuclear epitope to produce well-defined immunofluorescent staining patterns with minimal autofluorescence... 

Examples of typical staining patterns are shown in Fig 2 These profiles are analyzed for positivity by setting a lower limit of detection based on the isotypic control sample In our laboratory, we would normally set a region on the control that includes < 1 % of positive events and transpose this region to the M Ab-stained samples. 

Make a permanent record of the staining pattern by photocopying or photographing the blot. Alternatively, mark the positions of the bands of interest directly on the blot by overlaying the wet blot with plastic wrap or a plastic bag and using a pencil or pen to outline the bands. Press with moderate pressure to make a permanent indentation... 

Hirai, H. (1988) Paragonimus ohirai identification of nucleolar organizer regions (NORs) and silver nitrate staining pattern in spermatogenesis. Experimental Parasitology 67, 281-286. 

Fig. 24. Electronmicrograph of crystals formed from 2 molar CaClj extracts o( Mercenaria mercenaria following removal of salt. Negatively stained with Kf uranyl acetate (x 50000). A characteristic staining pattern is developed with well-defined periodicities. This material represents the MM. The crystals and staining pattern closely resemble tropomyosin-troponin paracrys-tals,93>...

Meek, K. M., Chapman, J. A., and Hardcasde, R. A. (1979). The staining pattern of collagen fibrils. Improved correlation with sequence data. /. Biol. Chem. 254, 10710-10714. 

Note that although immunohistochemistry is contributing significantly to clinical information, in the absence of quantitation, this method is subjective and prone to intra- and interobserver variations. To assure the reproducibility of histopathological results and then-correct evaluation, at least an assessment of the percentage of positive cells and both the staining pattern and intensity must be made. If available, computer-assisted image analysis... 

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