Fluorescence & UV- Visible Workshop Simple Applications to Sophisticated Analyses Why UV-Vis and Fluorescence? Quantitative measurements in solutions and solids Quality assurance and quality control (QA/QC) Sensitivity to the ppm level Full spectrum data for troubleshooting samples and method development Performance of windows and optics Diffuse and specular reflectance Integrating sphere detectors for scattering and non-planar samples Probe the electronic structure of organic and inorganic solids Fluorescence spectroscopy pyextends capability Measure your sample, not the matrix Linear measurements from percent to ppb Complimentary information to UV-Vis and FT-IR 2
Fluorescence Spectroscopy High resolution and sensitivity provide a new window to your samples Benefits of Fluorescence Spectroscopy Fluorescence is by far the most sensitive of all the molecular spectroscopy techniques Dramatically lower background compared to absorbance measurements Only technique with the ability to move from ensemble averaging to single-molecule analysis 4
Hardware Overview Double monochromator Excitiation light Emission light High resolution 0.5 nm spectral bandwidth Coated optics and holographic gratings increase sensitivity Fast scanning drive Stable and powerful xenon lamp Accessories that enable easy sample measurements Software modules customized for your application 5 Extreme Sensitivity Lumina detect concentrations as low as 60 femtomolar This is around 100 molecules! Low background provides a large linear range for concentration measurements Small volume cells allow low volumes to be measured Fluoresc cence Concentration Fluoresce ence 500 525 550 575 Wavelength 6
3D Scanning Provides Molecular Fingerprints By collecting many emission spectra, 3D fluorescence maps can be assembled 3D maps can identify: Hydrocarbon samples in water Drugs of abuse Dyes, inks, and additives Luminous software has powerful data visualization tools 7 Powerful 3D Graphics 8
Fluorescence Characterizes Solid Samples Fast changes (~ 20 µs) in luminescence intensity can be measured with the Lumina Optical components used in OLEDs and high-resolution h displays need this data to ensure a crisp, clear picture is produced OLED displays 9 UV-Visible Spectroscopy For Materials Science
Transmittance Optics Plano Pieces at normal or variable angle 11 Solar Cell and Display Windows Measure transmittance from 190nm to 1100nm at all viewing or illumination angles Fast, precise, simple 12
Transmittance Optics Compound or curved optics with the Diffuse Transmittance Accessory Magnetic base-plate drilled and scored like a laser table Integrating sphere detector is immune to changes of beam shape and focus 13 Specular Reflectance Optics How reflective is a mirror? Does it reflect in the UV? How effective is the AR coating on a piece of glass? Does it give true color? Is one mirror brighter than another Cold mirror lighting reflector 14
Application Focus - Dental Mirrors Red is rhodium dental mirror Green is new dental mirror Black is a reference mirror specified at 91% absolute reflectivity at 500 nm Measurements done on 30 reflectance accessory 15 Specular Reflectance Measurements Evolution 220, 300, 600 High precision and accuracy High resolution with SBW to 0.2nm Large variety of other accessories Evolution Array Convenience and speed VeeMAX 60 Spec Cold Mirror 16
Suspensions and Scattering Solids Most efficient small sphere on the market ISA-220 for Evolution 200 Cuvettes up to 50mm pathlength Scattering plastics Excellent performance in a low cost system 17 Diffuse Reflectance Integrating Spheres DRA-EV-600 and Evolution 600 Research standard double beam with red sensitive PMT detector Best sensitivity Integrating sphere dynamic range to > 3A 220-850 nm ISA-220 and Evolution 220 Exceptional performance for every-day measurements and basic research Unique matching of port size to beam Selectable 15nm SBW materials slit for outstanding performance with low reflectance samples 18
Small Volume Powder Measurements The Praying Mantis DRA uses Al optics and can measure down to 190nm Ideal for band gap studies at very short and volumes Small sample cup holds 0.021 cm 3 Large sample cup holds 0.236 cm 3 19 Band-gap Measurement on Semiconductor Powders 20
Oxidation Reaction Monitoring Prof. Huifang Xu, University of Wisconsin Madison Band Gap of Metal Complex Fe(III)-based Nano-plate Sample These features not visible in solution spectra Partially Reduced Reaction Product 21 Evolution 300/600 Praying Mantis Catalysis Systems UV-Vis Reflectance studies Coking Build-up of carbon on the surface Reduction of the surface Electronic structure of the surface of the catalyst Same Praying Mantis accessory used for UV-Vis and FTIR Only change the baseplate and optical windows! 22
Evolution 300/600 Praying Mantis Catalysis Systems 0.6 0.5 0.4 0.3 0.2 01 0.1 Iron loaded on zeolite Dilute sample with BaSO 4 (to bring on-scale) S4 1:3 and 1. Peak grind to homogenize oge e 2. Peak Pre-treat at 600 C in He to 3. Peak reduce all 5. iron Peak to Fe 2+, then cool and measure 4. Peak Summe Graph shows proposed deconvolution of peaks using off-line software 0 200 300 400 500 600 700 800-0.1 23 Evolution 300 Evolution 600 Double beam with Dual Silicon detectors Dynamic range to 4+ Absorbance Variable slits from 0.50 to 4.0nm 190-1100 nm Long lifetime Xenon flash lamp light source Walk-up measurements with no warm-up High intensity, especially in the UV Reliable with long time between qualifications Optional built-in Mercury lamp Calibration and verification on 8 emission lines Independent calibration on every slit Double beam with photomultiplier tube detector Dynamic range to 5.5 Absorbance Variable slits from 0.20 to 4.0nm 190-900 nm Stable W/D 2 continuous sources Optional built-in Mercury lamp Dual-wavelength calibration curve 24
Evolution Array UV-Vis Spectrometer Evolution Array UV-Vis Photodiode array technology Full spectrum (190-1100 nm): <1 second Simple, robust design Mechanical shutter is the only moving part Maximize up-time 10,000 hour tungsten lamp Open sample compartment Easy access to samples Easy to install accessories Stray light immunity 26
Full-Spectrum Analysis Full-Spectrum on Every Sample, Every Time Never need to recollect data because you chose the wrong wavelength Change analysis wavelength AFTER data is collected Improved troubleshooting of out-of-specification samples Failing samples must be investigated to determine the cause Spectrum holds more information than a single wavelength measurement Analytical Method Development 27 Evolution 200 Series UV-Vis Spectrophotometers Evolution 201, 220 and Evolution 260Bio
Evolution 200 Series New Double beam long lifetime Xenon system with Dual Silicon detectors Fast scanning double beam at 6000 nm/min Fast Kinetics at 100 data points/sec AFBG technology- Application Focused Beam Geometry Match resolution to the application Local control option with color touch screen Optional Mercury lamp Wavelength calibration and verification New generation INSIGHT software New generation Customized User Environment (CUE) Create dedicated application analyzers using local language prompts Patent pending 29 INSIGHT and CUE: A Revolution in Software Re-defined applications layered so the commonly used tasks are straightforward Intuitive graphics and setup wizards guide user through traditional UV-Vis applications Redefining Traditional UV-Vis Applications in QA/QC CUE creates completely customized interface giving you the simplicity of a dedicated analyzer for your measurements 30
What is CUE? Customized User Environment software A tool for customizing the user interface to run routine methods through the creation of scripts Script = set of actions that tells the software what to do and how to do it using a workflow diagram Automates and simplifies analyses by transforming multistep workflows into one easy-tofollow method 31 Cue in Action 32
Summary Quantitative measurement solutions for all sample types High sensitivity measurements and straightforward software for QA/QC High performance reflectance accessories for the optics industry Access to band-gap information in semiconductor materials FT-IR like convenience and speed for full spectrum analysis with the Evolution Array spectrophotometer p High specificity in fluorescence measurements on any sample 33