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 levely pp

• 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 extends capability

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p py p y• Measure your sample, not the matrix

• Linear measurements from percent to ppb

• Complimentary information to UV-Vis and FT-IR

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 tmeasurements

• Only technique with the ability to move from ensemble averaging to single-molecule analysis

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• Double monochromator• Excitiation light

• Emission light

Hardware Overview

• High resolution 0.5 nm spectral bandwidth

• Coated optics and holographic gratingsincrease sensitivity

• Fast scanning drive

• Stable and powerful xenon lamp

• Accessories that enableeasy sample measurements

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easy sample measurements

• Software modules customized for your application

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

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500 525 550 575

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Wavelength

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Concentration

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 datavisualization tools

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Powerful 3D Graphics

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Fluorescence Characterizes Solid Samples

• Fast changes (~ 20 µs) in luminescence intensitycan be measured with the Lumina

• Optical components used in OLEDs and hi h l ti di l d thi d t thigh-resolution displays need this data toensure a crisp, clear picture is produced OLED displays

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UV-Visible Spectroscopy For Materials Science

Transmittance Optics

Plano Pieces at normal or variable angle

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Solar Cell and Display Windows

• Measure transmittance from 190nm to 1100nm at all viewing or illumination angles

• Fast, precise, simple

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• 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

Transmittance Optics

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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?

D it i “t ” l ?• Does it give “true” color?

• Is one mirror “brighter” than another

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Cold mirror lighting reflector

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

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Measurements done on 30° reflectance accessory

Specular Reflectance Measurements

• Evolution 220, 300, 600• High precision and accuracy

• High resolution with SBW to 0.2nm

Large variety of other accessories• Large variety of other accessories

• Evolution Array• Convenience and speed

VeeMAX 60° SpecCold Mirror

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Suspensions and Scattering Solids

• Most efficient small sphere on the market

• ISA-220 for Evolution 200

• Cuvettes up to 50mm• Cuvettes up to 50mm pathlength

• Scattering plastics

• Excellent performance in a low cost system

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Diffuse Reflectance – Integrating Spheres

DRA-EV-600 and Evolution 600

• Research standard double beam with red sensitive PMT detector

• Best sensitivityBest sensitivity

• Integrating sphere dynamic range to > 3A

• 220-850 nm

ISA-220 and Evolution 220

• Exceptional performance for every-day measurements and basic research

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measurements and basic research

• Unique matching of port size to beam

• Selectable 15nm SBW materials slit for outstanding performance with low reflectance samples

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 cm3

• Large sample cup holds

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Large sample cup holds 0.236 cm3

Band-gap Measurement on Semiconductor Powders

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Oxidation Reaction MonitoringProf. Huifang Xu, University of Wisconsin – Madison

Fe(III)-based Nano-plate Sample Band Gap of Metal Complex

These featuresnot visible insolution spectra

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Partially Reduced Reaction Product

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

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Same Praying Mantis accessory used for UV-Vis and FTIROnly change the baseplate and optical windows!

Evolution 300/600 Praying Mantis Catalysis Systems

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S4 1:3

1. Peak

Iron loaded on zeolite• Dilute sample with BaSO4

(to bring on-scale) and grind to homogenize

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0.4 2. Peak

3. Peak

5. Peak

4. Peak

Summe

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• Pre-treat at 600°C in He to reduce all iron to Fe2+, then cool and measure

• Graph shows proposed deconvolution of peaks using off-line software

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200 300 400 500 600 700 800

Evolution 300 Evolution 600

• Double beam with Dual Silicon detectors

• Dynamic range to 4+ Absorbance

• Variable slits from 0.50 to 4.0nm

• Double beam with photomultiplier tube detector

• Dynamic range to 5.5 Absorbance

• Variable slits from 0.20 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

• 190-900 nm

• Stable W/D2 continuous sources

• Optional built-in Mercury lamp

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• Optional built-in Mercury lamp• Calibration and verification on 8

emission lines• Independent calibration on every slit

Dual-wavelength calibration curve

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 partMechanical 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

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Stray light immunity

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 collectedg y g

• 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

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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

N i C i d U E i (CUE)

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• New generation Customized User Environment (CUE)• Create dedicated application analyzers using local

language prompts

• Patent pending

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

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• CUE creates completely customized interface – giving you the simplicity of a dedicated analyzer for your measurements

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

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• Automates and simplifies analyses by transforming multi-step workflows into one easy-to-follow method

Cue in Action

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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

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• High specificity in fluorescence measurements on any sample