SPHERO TM Fluorescent Particles

Transcription

1 The SPHERO TM fluorescent microparticles are prepared by either staining polystyrene particles with a fluorophore solution or by polymerizing a fluorophore in styrene in the presence of polystyrene core particles. As a result, a wide variety of fluorescent particles can be prepared ranging in size, type of fluorophore, fluorescence intensity and surface functional groups. The fluorophores chosen for use in the preparation of SPHERO TM fluorescent particles are water insoluble and therefore are very stable. These fluorophores, once incorporated into the particles, do not leach and their color and fluorescence remains stable for long periods of time under proper storage conditions. The excitation and emission spectra of some of the fluorophores used in the SPHERO TM fluorescent particles are shown in Figure 16. Figure 16 Excitation & Emission Spectra of the fluorophores used in SPHERO TM fluorescent particles. SPHERO TM Fluorescent Beneficial to bioimaging and biosensing applications Uniform and stable fluorescence Available with functional groups for covalent binding of antigens or antibodies. UV Light Nile Red Pink Purple Nile Blue Sky Blue The SPHERO TM fluorescent particles are available in single or multiple fluorophores of various sizes and fluorescence intensities with very small coefficient of variation in both size and fluorescence. They can be used for latex agglutination, fluorescence microscopy, confocal fluorescence microscopy. Many of these particles can be used for flow cytometry. The flow cytometer histograms of 2.9 μm Nile Red (Catalog # FP ) at four channels are shown in Figure 17. More flow cytometry data for SPHERO TM fluorescent particles is shown on page 14. Figure 17 Flow cytometry histograms of 2.9 μm, Nile Red (Cat. No. FP ) Excitation (nm) UV Light Nile Red Pink Nile Blue Purple Sky Blue Emission (nm) NOTE: All spectra are taken with fluorescent particles in aqueous suspension.

2 The flow cytometry data of some of the fluorescent particles are shown in Table 1. Table 1 Fluorophores Cat. No. Size (µm) %CV* SPHERO TM Fluorescent, µm UV, Low FL UV, High FH Light, Low FL Violet 1 Violet 2 FL PE TR PE-Cy5 PE-Cy7 APC APC-Cy7 Light, High FH , Low FL , High FH Nile Red, Low FL Nile Red, High FH Pink FP Purple, Low FL Purple, Mid-level FP Purple, High FH Sky Blue FP SPHERO TM Fluorescent, µm FP Nile Red FP Nile Blue FP Blue FP SPHERO TM Multiple Fluorophore, µm UV/LY FP PR/Y, Low FL PR/Y, Mid-level FP PR/Y, High FH SPHERO TM Multiple Fluorophore, µm UV/LY FP PK/Y FP PR/Y FP SPHERO TM Fluorescent, µm FP Pink FP Purple FP *Data for particles with sizes above 1.0 micron obtained using a Dako Cyan ADP with the following Excitation and Emission wavelength: Violet 1: Ex 405nm, Em. 450/50nm; Violet 2: Ex. 405nm, Em. 530/40nm; FITC: Ex. 488nm, Em. 530/40nm; PE: Ex. 488nm, Em. 575/25nm; TR: Ex 488nm, Em. 613/20nm; PE-Cy5: Ex 488nm, Em. 680/30nm ; PE-Cy7: Ex 488nm, Em. >750nm ; APC: Ex. 633nm, Em. 665/20nm; APC-Cy7: 633nm, Em.> 750 nm **Data for particles with sizes below 1.0 micron obtained using a Stratedigm S1400 with the following Excitation and Emission wavelength: FITC: Ex. 488nm, Em. 530/30nm; PE: Ex. 488nm, Em. 545/60nm; TR: Ex 488nm, Em. 615/30nm

3 Figure 18 Microscope photos (a) 10 μm Nile Red beads (40x ), (b) 0.4 μm Avidin fluorescent beads binding to the surface of 6.0 μm Biotin polystyrene beads (5000x). (a) (b) SPHERO TM Fluorescent Polystyrene FP ml Nile Red FP ml Pink FP ml Purple FP ml Sky Blue FP ml Light FP ml FP ml Nile Red FP ml Purple FP ml Sky Blue FP ml Light FP ml FP ml Nile Red FP ml Pink FP ml Purple FP ml Sky Blue FP ml Light FP ml FP ml Nile Red FP ml Pink FP ml Purple FP ml Blue FP ml Sky Blue FP ml FP ml Light, Medium FP ml Nile Red FP ml Pink, Medium, FP ml Purple, Medium FP ml Nile Blue FP ml Blue FP ml Sky Blue FP ml FP ml Nile Red FP ml Pink FP ml Nile Blue FP ml Blue FP ml Ocean Blue FP ml Sky Blue FP ml FP ml Nile Red FP ml Pink FP ml Purple FP ml Sky Blue FP ml FP ml Nile Red FP ml Pink FP ml Ocean Blue FP ml FP ml Sky Blue FP ml UV FP ml Light FP ml FP ml Nile Red FP ml Purple FP ml PAK Blue FP ml Sky Blue FP ml UV FP ml Light FP ml FP ml Nile Red FP ml Purple FP ml PAK Blue FP ml Sky Blue FP ml FP ml Nile Red FP ml FP ml Nile Red FP ml Purple FP ml

4 SPHERO TM High Fluorescent Polystyrene SPHERO TM Low Fluorescent Polystyrene UV FH ml Light FH ml FH ml Nile Red FH ml Purple FH ml Sky Blue FH ml UV FH ml FH ml Nile Red FH ml Purple FH ml Purple FH ml SPHERO TM Fluorescent PMMA FPMA ml Nile Red FPMA ml Purple FPMA ml Nile Red FPMA ml Purple FPMA ml SPHERO TM Multiple Fluorophore Fluorescent Multiple Fluorophore FP ml Multiple Fluorophore FP ml Multiple Fluorophore FP ml Multiple Fluorophore FP ml Multiple Fluorophore FP ml Multiple Fluorophore FP ml UV / Light FP ml UV / Light FP ml UV / Purple / / Pink / Nile Blue Purple /, Medium Itensity FP ml FP ml UV / Light FP ml Pink / FP ml Nile Red / Blue FP ml Purple / FP ml Nile Red / Blue FP ml Purple /, High Purple /, Low FH ml FL ml FL ml UV FL ml Light FL ml FL ml Nile Red FL ml Purple FL ml Blue FL ml Blue FL ml Sky Blue FL ml Nile Red FL ml Blue FL ml Nile Red FL ml SPHERO TM FITC Polystyrene Surfaced labeled with FITC Used as calibration particles for flow cytometry Also used to cross calibrate different flow cytometers for data normalization. FITC F1CP ml FITC F1CP ml FITC F1CP ml FITC F1CP ml FITC F1CP ml FITC F1CP ml Figure 19 Flow cytometry histograms for Cat. No. FH lot AC01 (Fluorescent,, High, 1% w/v, 1.84 µm, 2 ml).

5 SPHERO TM Functionalized Fluorescent Used for covalent coupling of proteins or ligands Can be coated with Avidin, Biotin, Protein A, Goat anti-mouse IgG, or other protein of interest Used as fluorescent tracers for cell surface markers in fluorescence microscopy and flow cytometry. SPHERO TM Amino Fluorescent AFP ml Nile Red AFP ml AFP ml Pink AFP ml AFP ml Nile Red AFP ml Pink AFP ml Purple AFP ml Nile Blue AFP ml AFP ml SPHERO TM Carboxyl High Fluorescent CFH ml Nile Red CFH ml Pink CFH ml Nile Blue CFH ml SPHERO TM Carboxyl Fluorescent CFP ml Nile Red CFP ml Pink CFP ml Sky Blue CFP ml CFP ml Nile Red CFP ml Pink CFP ml Purple CFP ml CFP ml Nile Red CFP ml Pink CFP ml Purple CFP ml Sky Blue CFP ml CFP ml Nile Red CFP ml Pink CFP ml Purple CFP ml Sky Blue CFP ml CFP ml Pink CFP ml Purple CFP ml Sky Blue CFP ml Light CFP ml CFP ml Nile Red CFP ml Pink CFP ml Nile Blue CFP ml Sky Blue CFP ml Nle Red CFP ml Nile Red CFP ml SPHERO TM Carboxyl Low Fluorescent CFL ml CFL ml Pink CFL ml Nile Blue CFL ml SPHERO TM Dimethylamino Fluorescent Purple DFP ml SPHERO TM Carboxyl Multiple Fluorophore Fluorescent UV / Light / CFP ml