H the corresponding concentrations of histidine (200 mM), groups of other amino acids (200 mM), ct DNA (50 mM) or BSA (50 mM). The emission spectra were recorded in the 555?30 nm range, after equilibration at 25uC for 5 min. Excitation wavelength = 365 nm.Cell CultureHeLa cells were maintained in minimum essential medium (MEM) supplemented with fetal bovine serum (10 ), penicillin (100 U mL21), streptomycin (100 mg mL21) at 37uC under a humidified atmosphere with 5 CO2.ConclusionsIn conclusion, we have presented the cytoplasmic permeable iridium(III) complex 1 as a phosphorescent dye for live and fixed cell imaging. Complex 1 shows a bright phosphorescence in living cells, and effectively enters and stains the cytoplasm. Given that the emission properties of metal complexes can be fine-tuned through modifications of auxiliary ligands, we envision that further improvements can be achieved in the application of luminescent iridium(III) complexes as cellular imaging probes.Luminescence ImagingFor colocalization imaging of living cells. The cells were washed with PBS, then incubated with 10 mM 18325633 of iridium complex in DMSO/PBS (pH 7.4, 1:99, v/v) for 10 min at 37uC, and then further incubated with Hoechst 33258 for another 20 min before imaging. For colocalization imaging of fixed cells. The cells were detached from the MNS culture and were fixed with 4 paraformaldehyde at room temperature for 20 min. After washing with PBS, the fixed cells were incubated with 10 mM of iridium complex in DMSO/PBS (pH 7.4, 1:99, v/v) for 10 min at 37uC, and then further stained with Hoechst 33258 for another 20 min. After washing with PBS, the coverslips were separated from the chamber, and the cells were mounted with 10 glycerol and sealed with nail varnish on a glass substrate.Materials and Methods MaterialsIridium chloride hydrate (IrCl3.xH2O) was purchased from Precious Metals Online. DMSO, L-alanine, L-arginine, Lasparagine, L-glutamine, L-glycine, L-isoleucine, L-lysine, Lphenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-glutamic acid, L-cysteine, L-methionine, Lhistidine, bovine serum albumin, and calf thymus DNA were obtained from Sigma (St. Louis, MO). Hoechst 33258 and cell culture reagents were purchased from Invitrogen (Carlsbad, CA).Synthesis of [Ir(C^N)2(H2O)2]OTf (1?, where OTf = trifluoromethanesulfonate)The following complexes were prepared using literature methods: [Ir2(ppy)4Cl2] [77], [Ir2(bzq)4Cl2] [77], [Ir2(phq)4Cl2] [78], [Ir(ppy)2(H2O)2]OTf [79], [Ir(phq)2(H2O)2]OTf [76], and [Ir(bzq)2(H2O)2]OTf [76].Author ContributionsDirected the research: DLM CHL. Conceived and designed the experiments: DLM CHL. Performed the experiments: HJZ WCF HYK. Analyzed the data: DSHC HJZ LHC. Contributed reagents/materials/ analysis tools: DLM CHL WFF CYW. Wrote the paper: DSHC WCF.
Serum amyloid P component (SAP) is a plasma glycoprotein. It is a member of the pentraxin superfamily of calciumdependent ligand binding lectin proteins. Another member is Creactive protein (CRP), the classical acute-phase reactant in humans [1,2]. These proteins have been highly conserved throughout vertebrate evolution. There is a considerable degree of sequence homology (51 ASP-015K identity, 66 homology) within the pentraxin family, although the proteins are functionally distinct [3,4]. The biological role of SAP is only partly known. It binds in a calcium-dependent way to molecular arrays such as DNA, chromatin, histones, and phosphoethanolamine-co.H the corresponding concentrations of histidine (200 mM), groups of other amino acids (200 mM), ct DNA (50 mM) or BSA (50 mM). The emission spectra were recorded in the 555?30 nm range, after equilibration at 25uC for 5 min. Excitation wavelength = 365 nm.Cell CultureHeLa cells were maintained in minimum essential medium (MEM) supplemented with fetal bovine serum (10 ), penicillin (100 U mL21), streptomycin (100 mg mL21) at 37uC under a humidified atmosphere with 5 CO2.ConclusionsIn conclusion, we have presented the cytoplasmic permeable iridium(III) complex 1 as a phosphorescent dye for live and fixed cell imaging. Complex 1 shows a bright phosphorescence in living cells, and effectively enters and stains the cytoplasm. Given that the emission properties of metal complexes can be fine-tuned through modifications of auxiliary ligands, we envision that further improvements can be achieved in the application of luminescent iridium(III) complexes as cellular imaging probes.Luminescence ImagingFor colocalization imaging of living cells. The cells were washed with PBS, then incubated with 10 mM 18325633 of iridium complex in DMSO/PBS (pH 7.4, 1:99, v/v) for 10 min at 37uC, and then further incubated with Hoechst 33258 for another 20 min before imaging. For colocalization imaging of fixed cells. The cells were detached from the culture and were fixed with 4 paraformaldehyde at room temperature for 20 min. After washing with PBS, the fixed cells were incubated with 10 mM of iridium complex in DMSO/PBS (pH 7.4, 1:99, v/v) for 10 min at 37uC, and then further stained with Hoechst 33258 for another 20 min. After washing with PBS, the coverslips were separated from the chamber, and the cells were mounted with 10 glycerol and sealed with nail varnish on a glass substrate.Materials and Methods MaterialsIridium chloride hydrate (IrCl3.xH2O) was purchased from Precious Metals Online. DMSO, L-alanine, L-arginine, Lasparagine, L-glutamine, L-glycine, L-isoleucine, L-lysine, Lphenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-glutamic acid, L-cysteine, L-methionine, Lhistidine, bovine serum albumin, and calf thymus DNA were obtained from Sigma (St. Louis, MO). Hoechst 33258 and cell culture reagents were purchased from Invitrogen (Carlsbad, CA).Synthesis of [Ir(C^N)2(H2O)2]OTf (1?, where OTf = trifluoromethanesulfonate)The following complexes were prepared using literature methods: [Ir2(ppy)4Cl2] [77], [Ir2(bzq)4Cl2] [77], [Ir2(phq)4Cl2] [78], [Ir(ppy)2(H2O)2]OTf [79], [Ir(phq)2(H2O)2]OTf [76], and [Ir(bzq)2(H2O)2]OTf [76].Author ContributionsDirected the research: DLM CHL. Conceived and designed the experiments: DLM CHL. Performed the experiments: HJZ WCF HYK. Analyzed the data: DSHC HJZ LHC. Contributed reagents/materials/ analysis tools: DLM CHL WFF CYW. Wrote the paper: DSHC WCF.
Serum amyloid P component (SAP) is a plasma glycoprotein. It is a member of the pentraxin superfamily of calciumdependent ligand binding lectin proteins. Another member is Creactive protein (CRP), the classical acute-phase reactant in humans [1,2]. These proteins have been highly conserved throughout vertebrate evolution. There is a considerable degree of sequence homology (51 identity, 66 homology) within the pentraxin family, although the proteins are functionally distinct [3,4]. The biological role of SAP is only partly known. It binds in a calcium-dependent way to molecular arrays such as DNA, chromatin, histones, and phosphoethanolamine-co.