Single staining

As a test of reproducibility of spot printing and staining, a 10 spot by 10 row array (100 spots total) were printed on three slides. All slides were stained in a single staining run, using the same secondary detection chemistry with DAB as a chromogen. While DAB cannot provide accurate photometric measurements, it can provide relative comparisons from one spot to another on the same slide. When the stained control slides were analyzed in this... 

Samples in tubes 2 and 3 (see table below) are single-stained standards that will be used for setting compensation. These should contain a substantial number of apoptotic and/or necrotic cells, so that subsets that are positively stained with annexin V-PE and/or 7-AAD may be readily recognized, and are best prepared from designated replicates of test samples that have been exposed to a high concentration of toxicant. 

As a negative control, the crossreactivities of the secondary antibodies may be checked by exchanging them in single-staining methods. 

Double staining of the fetal skeleton for bone and cartilage (versus single staining of bone only for pharmaceuticals). 

Approximately one half of the fetuses in each litter (i.e., every alternate fetus in the uterus) are eviscerated and fixed for skeletal examination. The skeletal examination is performed following maceration of the soft tissue with aqueous potassium hydroxide and staining of the skeleton. Single staining, of the ossified bones only, is performed for pharmaceuticals (see Chapter 16) and double staining, of the bone and cartilage is performed for chemicals (see Chapter 17). 

Analyze a PE single-stained tube, such as PE-CD8, using scatter gating to restrict the analysis to lymphocytes... 

Analyze a PE-Cy5 single-stained tube, again using scatter gating. 

It remains to be said (unfortunately) that the compensation values are valid only for a particular pair of fluorochromes with a particular set of filters and mirrors and with particular voltages applied to a particular set of PMTs. If any one of those elements is altered, the required compensation values will alter as well. In general, once compensation has been set using single-stained particles with a given experimental protocol (PMT voltages, filters, and so forth), compensation values between a given pair of fluorochromes should remain... 

Analyze (i.e. acquire data but do not sort) -50,000 unstained cells to create a gate containing yeast cells on the basis of forward scatter and side scatter properties, as well as to control for autofluorescence. (If compensation is needed, analyze the singly-stained cells and compensate for emission overlap (see Note 12))... 

In vitro staining is used to colour cells or structures that have been removed from their biological context. Certain stains are often combined to reveal more details and features than a single stain alone can provide. Combined with specific protocols for fixation, sample preparation and investigative methods such as fluorescence microscopy, these standard staining techniques are used as consistent, repeatable diagnostic tools. 

King, L., and Morrison, M. (1976). The Visualization of Human Erythrocyte Membrane Proteins and Glycoproteins in SDS Polyacrylamide Gels Employing a Single Staining Procedure, , 4na/. Biochem. 71 223-230. 

In order to minimize spectral overlap during a multicolor flow cytometry analysis, a compensation procedure before acquiring data is required. Use the FACSDiva software package to automatically calculate a compensation matrix from single-stained controls (CompBeads, BD immunocytometry system, 552843 see Table 4). Below, a brief guide for getting started ... 

Acquire all single-stained compensation controls. If necessary, adjust the PMTV gains for each detector starting from those defined by CS T procedure. 

Table 4 Single-Stained Controls Used for Compensation Procedure...

Figure 5.47 Design and modeling of DNA nanotubes. (I) Representation of two crossed-over double-strand DNAs forming the core of a tile, shown as a rectangle. (II) RE tile, which can bind only to itself (the colors of the terminal single stains show complementary sequences). (Ill) SE tile, which can bind to itself. Two cores form (IV) a diagonal lattice and (V) a perpendicular lattice. ...

The study of phase size and compatibility requires that the different phases be observed, distinct from one another, and that there is minimum distortion in the polymers. However, multiphase polymers often cannot be stained or sectioned uniformly. There are composite structures that are combinations of soft rubbers, coatings and oriented fibers which caimot be stained with a single staining agent and the sections may be deformed or distorted, limiting both observation and interpretation. The ebonite method was developed by Smith and Andries [111] to stain and uniformly harden polymers. 

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