Staining double

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...

Van der Loos CM, Teeling P. A generally applicable sequential alkaline phosphatase immunohistochemical double staining. I. Histotechnol. 2008 31 119-127. 

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. 

A collection of useful protocols for the simultaneous immunoenzyme staining of two or more tissue antigens is available on the websites http //www.protocol-online.org/prot/Immunology/ http //www.vectorlabs.com/Protocols/MLB.pdf and http //www.ihcworld.com/. The protocols given in this chapter are practiced in the authors laboratory. These methods fall into two main categories (a) simultaneous immunoenzymatic double staining, and (b) sequential immunoenzymatic double/ multiple staining. 

Basic protocol for simultaneous immunoenzymatic double staining using primary antibodies of two different species or two different IgG isotypes... 

Basic protocol for sequential immunoenzymatic double staining (Adapted from http //www.vectorlabs.com/ and http //www.ihcworld.com/ books/Dako Handbook.pdf)... 

Stripping Buffers for Sequential Immunoenzymatic Double Staining... 

Tidman N, Janossy G, Bodger M, Granger S, Rung PC, Goldstein G (1981) Delineation of human thymocyte differentiation pathways utilizing double staining techniques with monoclonal antibodies. Clin Exp Immunol 45 457 467... 

Fukuda, N., Tanaka, H., and Shoyama, Y. (2001). Double staining of ginsenosides by western blotting using anti-ginsenoside Rbl and Rgl monoclonal antibodies. Biol. Pharm. Bull. 24, 1157-1160. 

Vandesande F. 1983. Immunohistochemical double staining techniques. Immunohistochemistry. Cuello AC, editors. Chichester John Wiley Sons. 

Zebhe, L, Sallstrom, J. F., Hacker, G. W., Hauser-Kronberger, C., Rylander, E., and Wilander, E. (1994) Indirect and direct on situ PCR for the detection of human papillomavirus. An evaluation of two methods and a double staining technique. Cell Vision 1, 163-167. 

Qualitative analysis of intrahepatic distribution is possible with immunohistochemistry. With antibodies against the carrier or the carrier-bound drug this compound can be localized in liver sections [152],To identify the cell type(s) involved in the uptake, the sections can subsequently be double stained with markers for the different cell types. In the rat liver, the monoclonal antibodies HIS52 (anti-rat endothelial cell antigen-1 or anti-RECA-1), ED2, the combination of anti-desmin and anti-glial fibrillary acidic protein, and anti-aSMA are generally used to identify SECs, KCs, quiescent HSCs, and activated HSCs, respectively. 

Roshash I mentioned in passing that when you double stain for PDF, it is restricted to its original homes. There is no PDF expression in any of these ectopic locations. Therefore we don t know what connects from these locations to behavioural outputs. 

Stanewsky Did you do a double stain against ELAV (this is a protein expressed in all neurons), for example ... 

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). 

Certain guidelines (5-7) require or give as an option a double staining method for the fetal skeleton with Alcian Blue added to stain cartilage together with Alizarin for the ossified material. 

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