Sulforhodamine rhodamine

Four forms of amine-reactive rhodamine probes are commonly available. Two of them are based on the tetramethyl derivatives of the fundamental rhodamine structure, one is based on the sulforhodamine B or Lissamine derivative, and the last is the sulforhodamine 101 or Texas Red-type of derivative. All of them react under alkaline conditions with primary amines in proteins and other molecules to form stable, highly fluorescent complexes. 

Lissamine rhodamine B sulfonyl hydrazine is a hydrazide derivative of sulforhodamine B that can spontaneously react with aldehyde- or ketone-containing molecules to form a covalent,... 

Texas Red hydrazide is a derivative of Texas Red sulfonyl chloride made by reaction with hydrazine (Invitrogen). The result is a sulfonyl hydrazine group on the No. 5 carbon position of the lower-ring structure of sulforhodamine 101. The intense Texas Red fluorophore has a QY that is inherently higher than either the tetramethylrhodamine or Lissamine rhodamine B derivatives of the basic rhodamine molecule. 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 derivatives of this fluorescent probe among the best choices of labels for use in double-staining techniques. 

Texas Red sulfonyl chloride is the active halogen derivative erf sulforhodamine 101. This important derivative of the basic rhodamine molecule possesses dual aliphatic rings off the upper-ring nitrogens and sulfonate groups on the No. 3 and 5 carbon... 

Lissamine rhodamine B sulfonyl hydrazine is a hydrazide derivative of sulforhodamine B that can spontaneously react with aldehyde- or ketone-containing molecules to form a covalent, hydrazone linkage (Fig. 217). It also can be used to label cytosine residues in DNA or RNA by use of the bisulfite activation procedure (Chapter 17, Section 2.1). The resulting fluorescent derivative exhibits an excitation maximum at a wavelength of 556 nm and a maximal emission wavelength of 580 nm when dissolved in methanol. In... 

Ujiie et al. [204] fabricated quartz chips for NCE and reported the separation of rhodamine B and sulforhodamine at 14.4 and 66.6 cm separator lengths. The buffer was 20 mM phosphate buffer at 2kV applied voltage and the separation was achieved in 70 seconds. Wakida et al. [205] reported a high throughput characterization for dissolved organic carbon in environmental waters within 2 minutes using NCE. The authors collected water samples from 10 sampling points at the Hino River that flows into Lake Biwa. Shin et al. [206] described NCE (PDMS) with fluorescence detection for analyses of atrazine. 

Rhodamines. The luminescence quantum yield study for molecules adsorbed on surfaces, which started with rhodamine 6G and 101 [81a], was later extended to zwitterionic rhodamines (sulforhodamine 101 and B) and to other nonrigid rtiodamines (rhodamines B and 3B [7, 82]. Important conclusions arising from those studies are that apart from rhodamines 6G and 101, only sulforhodamine... 

Sulforhodamine 101 and rhodamine 6G were further used for fluorescence quantum yield determination on different silica surfaces [7]. Silicas with controlled pore (22, 25,40, 60, 100, 150 A), and particle sizes were used. A system-... 

Sulforhodamine 101 and Rhodamine 6G Adsorbed on Different Pore Size Silicas... 

XPS experiments were also performed with sulforhodamine 101 and rhodamine 6G on silicas. The dye loadings under study were 0.002,0.025,0.10, and 1.0 fimdl/g onto silica gel with 25 A pores, and 0.002,0.04,0.10, and 1.0 /rmol/g... 

Fig. 50. Remission function values for sulforhodamine 101 and rhodamine 6G adsorbed on silicas with 22, 60, or 150 A pore diameters for the range of concentrations under study 1—0.004 mol/g to 8—0.1 iimoVg. Spectra are normalized to the maximum of the absorption of the dye. (All samples are Type I.)...

Fig. si. Atomic ratio N/Si for sulforhodamine 101 (full symbols) and rhodamine 6G (open symbols) adsorbed on 25 or 150 A pore silicas as a function of dye concentration. Straight lines in the plot represent average values. 

These studies indicate that sulforhodamine 101 forms nonplanarconformersin small pore size silicas as compared to large pore silica samples where the amount of conformers being formed is reduced. Rhodamine 6G samples exhibit very little conformer formation but their are still slightly dependent on pore size. Both rhodamines exhibit smaller fluorescence quantum yields when compared to the case of adsorption onto microcrystalline cellulose, this effect being more relevant in the sulforhodamine 101 case. 

Figure 5. Calculated signal gain versus signal wavelength for Rhodamine B (RB), Rhodamine 6G (R6G), DCM, Rhodamine 101 (RlOl), and Sulforhodamine 101 (SlOl)-doped GIPOFA. Launched signal power (FWHM = 3.5 ns) = 1.0 W. Launched pump power (at 532 nm, FWHM = 6.0 ns) = 10 kW. Core diameter = 500 [Am.

Sulforhodamine B 2 ethyl groups on each nitrogen on the outer ring plus a sulfonate at positions 3 and 5 (Lissamine rhodamine B)... 

The extensive rhodamine dye family is based on the xanthene structure (see Fig. 12-7) (Schultheiss, 2002). One of the most common rhodamine dyes is rhodamine 6G (R6G), also called rhodamine 590, shown in figure 12-8. The excitation and emission spectra of R6G are shown in Figure 12-9. Rhodamine 6G has been found to be an efficient and stable dye. The rhodamine family is created by varying the substituents, especially of the amino groups which appear at positions 3 and 6. of particular interest is sulforhodamine 101... 

In principle, any couple of fluorophores can be used for FRET, provided that the emission spectrum of the donor overlaps with the absorption of the acceptor. For a review of FRET-couples (and RO values) of chemical dyes see [62]. Furthermore, donors with a high fluorescence quantum-yield and acceptors with a high molar absorbance will display increased FRET. For FLIM it will be important to tune the instrument-performance to ensure maximal sensitivity to small changes in lifetimes at the control donor lifetime. Usually this is easily achieved. Many FRET-pairs have been used for FRET-FLIM including chemical probes as Fluorescein-Rhodamine [54,93],calcein-sulforhodamine B [94], and Cy3-Cy5, [70]. Since 1996, the availability of genetic-encoded fluorophores such as CFP, GFP, YFP has boosted application of FRET-FLIM enormously [95]. Nowadays fluorescent-tagging of proteins no longer depends on laborious protein pu-... 

---