Spectrofluorophotometer RF-5301PC C125-E008J
World s Highest Level Performance and Windows Software Open Up a New Spectrum of Fluorescence Analysis. High-Sensitivity (SN Ratio Over 150) Sensitivity is the essence of fluorescence analysis. The high-throughput optical system in the RF-5301PC employs a blazed holographic grating, photomultiplier and digital circuit to provide the highest level SN ratio attainable. High-Speed Scanning High-speed scanning up to 5,500 nm/min allows you to measure a spectrum in seconds. In addition, a monochromator slewing speed of about 20,000 nm/min enables quick and easy setting of two or more wavelengths. Versatile Sample Compartment Size The sample compartment measures 140 mm wide, 170 mm deep and 140 mm high, enabling the use of micro cells, high-sensitivity cells, LC flow cells, etc. for a wide range of applications. Built-In Performance Checks When the instrument is switched ON, operating conditions of the spectrophotometer are automatically verified. In addition, built-in noise level (SN ratio) and light source (Xenon lamp) usage features help maintain the instrument in its optimum condition, providing absolute confidence in the quality of the data.
Windows-Based Workstation A PC directly controls the instrument for data acquisition and processing. The Windows friendly operating environment allows you to perform measurement, data processing, editing and recording in one continuous operation with a click of the mouse. The Copy Graph function allows easy transfer of measurement data or spectra to word processing or spreadsheet software for preparation of documents or additional calculations. From Routine Analysis to Research Employing the Highest Level of Sensitivity in the World! Features of Fluorescence Spectrometry Tens to thousands of times higher sensitivity than absorption spectrometry High selectivity (combining excitation and fluorescence wavelengths allows measuring even mixtures) Life Sciences Amino acids, peptides, proteins, fats, sugars, vitamins, coenzymes, amines, etc. Qualitative/quantitative analysis Enzyme reaction research Research of reaction mechanisms through analysis of substrates, coenzymes, and proteins (tryptophan and tyrosine residue) Antigen-antibody reactions Research of protein-protein, protein-nucleic acid, and protein-sugar interactions Analysis of intracellular ions (such as Ca 2+ ) using a fluorescence probe (such as Fura-2) Analysis of neurotransmitters (such as catecholamines) Research of nucleic acid hybridization Research of biomembranes Food Products Amino acids, proteins, fats, carbohydrates, vitamins, coenzymes, antibiotics, food additives (such as flavorings, seasonings, and antioxidants), cooking oil (such as vegetable oils and animal oils), and pesticides (carbamate, organophosphorus, and 1-naphthylacetic acid) Food ingredient analysis and quantitative analysis Inspections for residual pesticides Chemicals / Environment Organic, inorganic, and polymer compounds (such as fibers, plastics, and rubbers) Spectrofluorophotometer RF-5301PC Trace analysis Materials science research Photochemical reaction research Polymer research Optical materials research Assessment of atmospheric, water, and soil pollution such as the analysis of polycyclic aromatic hydrocarbon compounds Crude oil and coal analysis
Windows-Based Software Provides Powerful Support for Sample Analysis and Data Processing Windows-Based Control 3 Measurement Modes SpeedBox TM Select from 3 measurement modes Copy and Paste Wide Selection of Data Processing Functions Enables comprehensive management, from measurements to data analysis and report creation/output. Online help is provided as well. According to the Sample Spectrum Measurement Mode To measure spectra, simply click the [Start] button. Displays up to 10 spectra in the same window and performs calculations interactively. Freely zoom in or out using the mouse. Shimadzu's proprietary radar function allows viewing all spectra with a single click. Quantitative Measurement Mode Automatically creates calibration curves from measurement of standard samples. To quantitate unknown samples, simply click the [Read] button. PopUpScan allows confirming spectra while quantitative measurements are still in progress. Quantitation method: Peak height Calibration curves: 1st to 3rd order equations Mean value calculation function Time Course Measurement Mode High sensitivity, a stable baseline, and a 20-bit high-resolution A/D converter ensure reliable detection of small changes in samples. The activity value calculation function supports even enzyme research. 4
One-Click Operation SpeedBox Create buttons for frequently used functions to execute the functions with a single click. Freely customize button shapes, locations, and other features. Compatible with Commercial Software Applications Copy & Paste Data can be copied and pasted to Windows-based applications, such as word processing and spreadsheet software, which allows creating sophisticated reports. The ASCII conversion function allows transferring data to spreadsheet software. Simple and Powerful Extensive Data Processing Functions Use these data processing functions to correct data, extract hidden information, and so on, from the spectra obtained. Four basic arithmetic operations Smoothing Area calculation Differentiation Peak detection Activity value calculation Operate all functions easily and visually using the mouse. RF-5301PC Spectrofluorophotometer 5
Fast and Smart Monochromator and High Expandability Provide Powerful Support for Measurement and Analysis SN Ratio over 150:1* Max. Scan Speed of 5,500 nm/min Wavelength Slewing Speed of 20,000 nm/min Search Function Self-Diagnostic Function High sensitivity and high speed scanning provide quick and easy measurements. The search and self-diagnostic functions allow performing measurements without time-consuming processes. * Signal-to-noise ratio is from a Raman peak for distilled water with an excitation wavelength of 350 nm and bandwidth of 5 nm. High Sensitivity SN Ratio over 150:1 A bright optical system featuring a blazed holographic grating and digital signal processing enable ultra-trace analysis at signal-to-noise ratios over 150:1. Note: SN ratio is measured for the Raman line of distilled water Excitation wavelength Bandwidth 0-98 % response 350 nm 5 nm 2.0 sec Detects Optimal Excitation and Fluorescence Wavelengths Search Function Simply click the [Search] button. Automatically detects optimal excitation and fluorescence wavelengths. Automatically determines fluorescence from Rayleigh scattering, Raman scattering, 2nd-order light, etc. Makes time-consuming wavelength settings easy. Note: For a bandwidth of 10 nm, the SN ratio is equivalent to over 600:1. Enables High-Speed Measurements Max. Scan Speed of 5,500 nm/min An advanced direct wavelength drive mechanism provides spectra with minimal distortion, even at high speeds. A maximum scan speed of 5,500 nm/min enables high-speed spectra monitoring. Fast 20,000 nm/min wavelength slewing speed improves measurement efficiency. Automatic Instrument Performance Management Self-Diagnostic Function Automatically checks the operating status of the drive system when power is switched ON. Excitation slit Excitation monochromator Fluorescence slit Fluorescence monochromator Zero setting The software automatically performs time-consuming instrument management and performance checks, making it possible to use the instrument in an optimal state at all times. Manages lamp usage hours. Automatically measures and calculates SN ratios. 6
Large Sample Compartment High Degree of Expandability Easy to Use, Large Size Large Sample Compartment Tailor systems to samples and measurement methods. Allows Configuring a Variety of Systems High Degree of Expandability A large sample compartment provides plenty of space, improving ease of use. A wide variety of optional equipment can be attached. Various types of cell holders Various types of flow cell units Polarizer attachment Outputs analog signals of fluorescence intensity. Can be connected to a Chromatopac integrator Enables chromatographic measurements Note: Analog signals are output at a full scale of 1 V. Systems can be configured to samples and measurement methods. Specifications Hardware Software Light source Excitation and emission monochromators Wavelength scale Measuring wavelength range Spectral bandwidth 150 W Xenon lamp. Ozone resolving type lamp housing Blazed holographic grating F2.5, 1300 grooves/mm. 220-990 nm and 0 order 220-750 nm and 0 order as standard. 220-900 nm and 0 order with the optional R928 photomultiplier 6-step selection of 1.5, 3, 5, 10, 15 and 20 nm. (6 nm bandwidth is available for half sample height on the excitation side only.) Measurement Data processing Filing Excitation, emission and synchronous spectrum measurement, time-course measurement, quantitation, automatic search of optimal excitation and emission wavelengths, PopUp Scan. Arithmetic calculation between spectrum and a constant, smoothing, 1st through 4th derivatives, 1/Y, logarithmic conversion, data printout (with or without activity value computation), peak pick, point pick, area calculation, averaging (in quantitation), generation of calibration curves of 1st through 3rd order Save, recall, and delete of data. Conversion into ASCll and DIF formats. Wavelength accuracy Light source compensation Sensitivity ±1.5 nm Dynode feedback system with monochromatic light monitoring The SN ratio is 150 or higher for the Raman line of distilled water (350 nm excitation wavelength, 5 nm spectral bandwidth, and 2-second response for 98% of the full scale). Data output Maintenance Automatic scale adjustment, readout of data at user-specified point, data printout (preview function provided), selection of colors and types of curves Automatic monitoring of signal-to-noise ratio, monitoring of the run time of light source lamp Wavelength scanning Wavelength slewing speed Response 7-step selection of Survey (about 5500 nm/min), Super (about 3000 nm/min), Very Fast, Fast, Medium, Slow and Very Slow About 20,000 nm/min. 8-step selection of 0.02, 0.03, 0.1, 0.25, 0.5, 2, 4 and 8 seconds for 98% of the full scale User interface Other functions PC requirements SpeedBox (assign icons for commonly used menu commands) Data exchange via clipboard, auto response control, automatic shutter CPU: 1GHz or higher OS : Windows 7 Professional, Windows XP Professional (SP2/SP3), Windows Vista Business (SP1) 32 bit Sensitivity selection 2 steps of HIGH and LOW (The sensitivity at HIGH is about 50 times that of LOW.) Interface RS-223C interface, interface for autosampler, and interface for sipper unit. Dimensions and weight W667 D530 H270 mm; 43 kg. Power requirements 100, 120, 220, 240 V; 50/60 Hz; 400 VA. Operational temperature range 15-35 C Operational humidity range 40-80% (Below 70% with temperature higher than 30 C). RF-5301PC Spectrofluorophotometer 7
Wide Variety of Optional Accessories Provides Powerful Support for Measurement and Analysis Intracellular Calcium Measurement System This measurement system uses a fluorescence probe to measure real-time changes in concentrations of Ca 2+, H +, and other intracellular messengers. Measures up to 4 wavelengths, allowing measurements using multiple fluorescence probes. High-speed switching between wavelengths enables switching from 340 nm to 380 nm in a minimum 0.5 seconds, using a Fura-2 probe. Displays real-time changes in fluorescence intensity (maximum of 4) and ratios (maximum of 4) during measurements. Support for quantitation using single-wavelength quantitation, two-wavelength quantitation, and calibration curves enables measurement using Fluo-3, Fura-2, BCECF, and other fluorescence probes. Compatible with Window 7 Professional / XP Professional (SP2 / SP3) and Vista Business (SP1). System Contents P/N RF-5301PC Spectrofluorophotometer Intracellular Ca 2+ Measurement Software (Super Ion Probe) Constant-Temperature Cell Holder with Stirrer Constant-Temperature Water Circulator Syringe Adapter Computer and printer P/N 206-24800-91 206-84838-92 206-24930-91 208-97263 206-69266-01 8
Options NTT-2200P Constant- Temperature Water Circulator P/N 208-97263 Constant-Temperature Single Cell Holder with Stirrer P/N 206-24930-91 (100V) P/N 206-24930-92 (120V) P/N 206-24930-93 (220/240V) High Sensitivity Cell Holder P/N 204-26841-01 Circulates temperature-controlled water to a constant-temperature cell holder. -Temperature range: Ambient + 5 C to + 80 C -Temperature control precision: 0.05 C~ -Max. pumping rate: 27/31 L/min, 9.5/13 m (50/60 Hz) -External circulation nozzle: 10.5 mm OD (both outlet and return) -Tank capacity: About 10 L (9 L during use) -Safety features: Detection of over-temperature of Upper or Lower limits, Detection of heater wire malfunction, Protection of heating too little circulating water, Detection of sensor malfunction, Independent overheat protection, Over current circuit protector -Standard accessories: Lid with handles, Rubber hose (4 m, 9 mm ID), Hose clamps (4 pc.), Instruction manual -Dimensions: W270 H560 D400 (mm) -Power requirements: 100 VAC, 1250 VA, with 1.7 m power cord and grounded plug Note: Piping to Constant-Temperature Cell Holder (P/N 206-24930-xx) is possible by the hose (9 mm ID). Keeps the sample at the desired constant temperature during measurement by circulating temperature-controlled water (5-70 C) around the sample cell. The stirring option has variable speed control and the minimum sample volume is 2 ml. Effective for fluorometry of suspended cell samples. Note: Does not include the Temperature Controlled Water Bath. Not applicable to samples having a specific gravity greatly different from that of the solvent. Note: The hose joint is applied to the 6-9 mm ID tube. Constant-Temperature Four Cell Holder P/N 206-24940-91 Keeps four sample cells at the desired constant temperature via connection to a water bath. Note: Requires the Temperature Controlled Water Bath. This cell holder enhances the sensitivity two to three times or more. This is recommended for low concentrations. It repeatedly reflects the excitation and emission beams through the sample to increase the excitation efficiency. Solid (Powder) Sample Holder P/N 204-26836-01 Provides a means to examine the fluorescence of a solid sample. The angle of the holder is designed to limit the reflected excitation beam from the emission monochromator. Standard accessories include a sample holder, quartz plate, powder sample plate, spacers and a sample fixing bar. Cutoff filters are included to reduce scattering (U-340, IHU-310, L-42, Y-50, O-56, R-60). Rectangular Flow Cell, 120µL P/N 204-03285-04 Used for fluorescence monitoring of effluents in column chromatography. Made of fused silica. Requires adapter (P/N 206-83934-91). RF-5301PC Spectrofluorophotometer 9
Options Polarizer Attachment (for UV/Vis region) P/N 204-03290 Polarizer Attachment (for Vis region) P/N 204-03290-01 Quartz Cell Polished on Four Sides P/N 200-34441 Non-Fluorescent Cell P/N 200-34594-03 Flow Cell Unit (120L) for LC Application P/N 204-06249 Measurement of fluorescence polarization provides information on the size, fluidity and environments of fluorescent molecules. Permits reliable measurement of polarization characteristics over a wide wavelength range from 240 nm to 800 nm. Micro Cell Holder with Micro Cell P/N 204-27125 Provides a 10 mm square non-fluorescent cell, polished on four sides. This cell is recommended for trace analysis, particularly when using low UV wavelengths for excitation. Does not fluoresce when excitation wavelengths from 240 to 260 nm are used. Flow-Thru Cell Unit for HPLC (12L) P/N 204-05566 The square micro-flow cell has an inner volume of 120 µl and is made of silica to ensure minimum light scattering. The inlet and outlet tubes are both made of vinyl resin and are 0.5 mm I.D. and 2 mm O.D. R-928-08 Photomultiplier P/N 200-75021 Permits measurement of samples as small as 400 µl in volume. The cell is installed in an adapter of the same dimensions as an ordinary 10 mm square cell, so it can be placed in the standard cell holder. With this attachment, the RF-5301PC is used as a highly sensitive spectral fluorescence monitor in HPLC. Free selection of the excitation and emission wavelengths permits selective detection. The mobile phase stream can be stopped for spectral measurement, so that chromatographic peaks can be qualitatively determined with higher reliability. The flow-thru cell is a square type with an inner volume as small as 12 µl and is made of silica, ensuring minimized light scattering. Extends the measuring wavelength range from 750 to 900 nm. Filter Set P/N 204-04691 Includes 8 filters: two broad-band interference filters U-340 and B-390 for isolation of excitation wavelengths of 300-380 and 350-450 nm, respectively. Six cutoff filters: UV-29, L-41, Y-46, Y-50, O-56 and R-60 with approximately 50%T at 300, 420, 460, 500, 560 and 600 nm, respectively. 10
8mm dia. Test Tube Holder P/N 204-05853 Bottom Rise Sample Stage P/N 204-04811 Sipper Unit 5300 P/N 206-24950-91 Holds a test tube, 8 mm outer diameter and 45 mm-100 mm long. The minimum sample volume required is 400 µl. Raises the sample cell position in order to reduce the minimum sample volume to 1.5 ml. Aspirates solution samples using a peristaltic pump driven by a stepping motor, to be measured successively. Minimum sample volume is 2 ml. Linked operation with the ASC-5 Auto Sampler possible. 12mm dia. Test Tube Holder P/N 204-03293 Spectra Correction Accessory P/N 204-07550 Used to correct excitation and emission spectra in the wavelength region from 250 nm to 600 nm, using rhodamine solution as the reference (quantum counter). ASC-5 Auto Sample Changer P/N 206-23810-xx Holds a test tube, 12 mm outer diameter and 60 mm-100 mm long. Builds an automatic multi sample spectrophotometry system in combination with a sipper unit. Up to 100 test tubes are set together on the rack. The aspirating nozzle is programmable to move X, Y and Z (vertical) direction. RF-5301PC Spectrofluorophotometer 11
RF-5301PC Company names, product/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation or its affiliates, whether or not they are used with trademark symbol TM or. Third-party trademarks and trade names may be used in this publication to refer to either the entities or their products/services. Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own. www.shimadzu.com/an/ For Research Use Only. Not for use in diagnostic procedures. The contents of this publication are provided to you as is without warranty of any kind, and are subject to change without notice. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. Shimadzu Corporation, 2013 Printed in Japan 3655-07317-30AIT