Rhodamine staining

Zijlmans JM, Visser JW, Kleiverda K, Kluin PM, Willemze R, Fibbe WE. Modification of rhodamine staining allows identification of hematopoietic stem cells with preferential shortterm or long-term bone marrow-repopulating ability. Proc Natl Acad Sci USA. 1995 92 8901-8905. 

The method allows visualization of the epithelial barrier and penetration of small molecules into cell layers indicating disruption of tight junctions (Saitou et al. 1998). Slides are examined under the fluorescent microscope for bound biotin and rhodamine staining in deeper epithelial and stromal layers. Staining indicates penetration of biotin molecules into the tissue through disrupted tight junction barriers (Xu et al. 2004). Recovery from disruption is tracked on day 0,1,2,3, and 4. 

Some specimens required either longer exposure times or greater concentrations of fluorescent dye to incorporate it throughout the hair. In only a Korean hair sample was uniform rhodamine staining throughout the hair shaft not achieved. 

Fig. 9.2 a Confocal image of rhodamine-stained enamel proximal lesions treated with resin infiltrant (Icon DMG). b Fissure caries lesions green) infiltrated with resin red) (color figure online). Reproduced with permission from [137] and [138]... 

Out of 440000 cells, 17 were capped" rhodamine-stained-POL-binding cells (Table 1). The fluorescein labeling coincided with the rhodamine labeling in 14 cells out of 17. In these cases, no fluorescence was detectable outside the cap. In the rest (3 out of 17) the fluorescein staining was most intense at the rhodamine cap but also could be seen on the rest of the cell surface. Further experiments showed that non-capped immunoglobulin on such cells had anti-POL specificity. This was determined by a double treatment with POL, first... 

POL a 26 ng/ml R-anti- POL Rd R-anti-c MIg FI Additional treatment or pretreatment Capped rhodamine staining (POL-binding cells) Fluorescein labeling coinciding with capped rhodamine staining... 

Total pectinase, cellulase and lipase activities secreted by colonies were detected on BSM plates containing respectively 1% of citrus pectin, 2% Walseth cellulose and 1% olive oil + rhodamine. After few days at 30°C, pectin plates were covered by 1% CTAB for Ihour, positive colonies became surrounded by a clear halo walseth plates are not stained the halo is visible directly on positive clones lipase activity is revealed under UV on oil-rhodamine plates. 

Microscopic examination ( smear ) detects about 8 to 10 X 1CF organisms/L of specimen using the older AFB (acid-fast bacillus) stain, with the newer auramine-rhodamine fluorsecent... 

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. 

FIGURE 24-1 Phosphotyrosine staining of a hippocampal neuron. A cultured rat hippocampal pyramidal neuron is stained with an antiphosphotyrosine antibody and detected by a secondary antibody conjugated to rhodamine. The staining reflects the presence of tyrosine-phosphorylated proteins throughout the neuron, including the cell body and synaptic regions. 

Mitochondria are distinct organelles with two membranes. The outer membrane limits the organelle and the inner membrane is thrown into folds or shelves that project inward and are called cristae mitochondriales. The uptake of most mitochondrion-selective dyes is dependent on the mitochondrial membrane potential. Conventional fluorescent stains for mitochondria, such as rhodamine and tetramethylrosamine, are readily sequestered by functioning mitochondria. They are, however, subsequently washed out of the cells once the mitochondrion s membrane potential is lost. This characteristic limits their use in experiments in which cells must be treated with aldehyde-based fixatives or other agents that affect the energetic state of the mitochondria. To overcome this limitation, the research... 

Rhodamine 6G (0.1 g l-1 in water) Spray or dip plate and view under UV light A versatile stain for all lipid classes. Pink-purple-red spots. Most useful with alkaline solvents. The dye may be incorporated in the plate... 

Figure 5 Epifluorescence microscopic images of BT 20 cells (A) Cells incubated with Rhodamine-PE labeled stearyl triphenylphosphonium liposomes (B) mitochondria in BT 20 cells stained with Mite Tracker Red. Abbreviation. PE, phosphatidylethano-lamine. Source Prom Ref. 30.

Wessendorf, M. W. and Brelje, T. C. (1992) Which fluorophore is brightest-A comparison of the staining obtained using fluorescein, tetramethylrhodamine, lissamine rhodamine, Texas Red, and cyanine 3.18. Histochem. 98, 81-85. 

Fig. 3. Comparisons of wide-field (A) and confocal fluorescence images (B, mesoglea level C, apical) of rhodamine phalloidin-stained F-actin in a whole-mount hydra tentacle. The hydra was fixed and stained as described in Chapter 18. The bar represents 25 pm. All images were collected with a Nikon (New York) Microphot FX microscope (x40 objective lens). Confocal images were collected with the microscope connected to a Bio-Rad (Hercules, CA) MRC600 laser-scanning confocal system.

Fig. 5. Optical sectioning of rhodamine phalloidin-stained F-actin in a neutrophil migrating through a 5-pm pore of a polycarbonate membrane. The neutrophil migration is stimulated in response to 10 M Af-formytmethionyl-leucy 1-phenylalanine. (A), (B), and (C) correspond to O.S-pm optical sections indicated as sections A, B, and C, respectively, in Fig. 4. The bar represents 10 pm. The images were collected with a Nikon Microphot FX microscope (x60 Plan-apochromat lens, numerical aperture, 1.6) connected to a Bio-Rad MRC600 laser-scanning confocal system.

The phallotoxins, phalloidin and phallacidin, are bicyclic peptides with mol wts of 789 and 847 Dalton, respectively. The NBD phallacidin and rhodamine phalloidin conjugates have been most frequently utilized in flow cytometric assays. The fluorescent phallotoxin conjugates have mol wts of 1000-1200 Dalton, are water-soluble, and stain actin at nanomolar concentrations (reviewed in ref. 19). Unconjugated phallotoxins should be obtained to verify the specificity of fluorescent phallotoxin staining to F-actin (see Note 4). 

Kim M, Cooper DD, Hayes SF, Spangrude GJ. Rhodamine-123 staining in hematopoietic stem cells of young mice indicates mitochondrial activation rather than dye efflux. Blood. 1998 91 4106-4117. 

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