Extension of NRC fluorescence measurement capabilities

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Ex t e nsion of N RC Fluore sc e nc e

M e a sure m e nt Ca pa bilit ie s

CORM 2012, Session I, 30 May 2012

NRC Sussex, Ottawa, Canada

Joanne Zwinkels, Mario Noel and Bill Neil

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Out line

• Background of NRC Fluorescence Project

- Instrumentation & services: surface fluorescence

• Extension to Volume Fluorescence

- Results of international comparison

• Extension to NIR Fluorescence

- Impact on use of colour standard tiles

• Extension to Other Measurement Geometries

- 3M Goniospectrofluorimeter Facility

- Preliminary characterization results

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N RC Re fe re nc e Spe c t rofluorim e t e r

• Based on two-monochromator method • Measurement geometry: 45°a/0°

J. Zwinkels et al., Applied Optics, 36, 892-902 (1997)

370 400 430 460 490 520 550 580 610 640 250 290 330 370 410 450 0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03 6.0E-03 7.0E-03 8.0E-03 L um in an ce r ad ian ce f act or Emission (nm) Excitation (nm) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 300 400 500 600 700 800 Wavelength (nm) R adi anc e f ac tor s Total D65 Reflected Luminescent

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Why Ca librat e ?

500 ₅₆0 620 ₆₈0 740 800 370 420 470 520 570 620 0.0E+00 2.0E-03 4.0E-03 6.0E-03 8.0E-03 L u m in an ce r ad ian ce fact o r Em ission (nm ) Excitation (nm ) Raw Bispectral Lum inescent Radiance Factor

500 ₅60 620 680 740 800 370 420 470 520 570 620 0.0E+00 2.0E-03 4.0E-03 6.0E-03 8.0E-03 1.0E-02 L u m in an ce r ad ian ce fact o r Em ission (nm ) Excitation (nm ) Norm alized Bispectral Lum inescent Radiance Factor

0.0 0.5 1.0 1.5

300 400 500 600 700 800

Raw “Measured” _{After Calibration}

Total (D65)

True result

“after calibration” Erroneous result “raw” data

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Surfa c e Fluore sc e nc e

N RC Ca librat ion Se r vic e s

Total Spectral Radiance Factors

β_T(45ºa/0º); β_T = β_R + β_F  300 – 850 nm, 5 nm bandpass • Selected illuminants (CIE D65, C,A, D50, custom)

• In accordance with ASTM and CIE

colorimetry standards

Solid Samples:

Paper, textile, paint, plastics, pressed powders

Spectral Quantum Yields

Fluorescent Quantum Yield

0% 10% 20% 30% 40% 50% 60% 70% 250 300 350 400 450 Excitation Wavelength (nm)

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Ex t e nsion t o Volum e Fluore sc e nc e

Background:

• Increasing use of fluorescence techniques as a quantitative

analytical tool

• Increase in biological and medical applications of fluorescence,

e.g. probes and labels

• Increasing diversity of fluorescence instrumentation

• Fluorescence measurements are very dependent on

instrument-specific effects (wavelength, polarization, aging)

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Ex t e nsion t o Volum e Fluore sc e nc e

Specifications for Intercomparison:

SAMPLES: fluorescent dyes

• Transparent

• Side-viewing

• 0°/90° geometry

• High resolution (1 or 2 nm)

• Low concentration (weak

fluorescence)

• LI QU I DS

Specifications of NRC Spectrofluorimeter:

IDEAL: fluorescent surface colours

• Opaque

• Front-surface viewing

• 45°/0° geometry

• Medium resolution (5 nm)

• Moderate to strong fluorescence

• SOLI DS

NRC – invited to participate in an International Comparison of Correction of Emission Spectra

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Pre lim ina r y N RC Re sult s

Spectrum of Quinine Sulfate Dihydrate, First look !

370 390 410 430 450 470 490 510 530 550 570 590 610 630 650 670 Wavelength (nm) R el at ive F lu o rescen ce I n ten si ty Expected value at HV= 1100 V

S/N ~ 5:1

=> Need to

Increase

S

and

Decrease

N

Increase S:

- Optimize sample presentation - Optimize system throughput

Decrease N:

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Ex t e nsion t o Volum e Fluore sc e nc e

Re-optimization of instrument parameters

- No ground loops - No cooler switching noise - Choice of cuvette - Cuvette positioning - PMT high voltage

S/N ~ 100:1

(20x

improvement

)

0 0.2 0.4 0.6 0.8 1 1.2 375 400 425 450 475 500 525 550 575 600 625 650 675 R el at iv e F lu o rescen ce I n ten si ty Wavelength (nm)

Quinine sulphate dihydrate (0.08A) Raw

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N RC Fina l Com pa rison Re sult s

0.0E+00 1.0E-04 2.0E-04 3.0E-04 4.0E-04 300 350 400 450 500 550 600 650 700 750 A B C D E Y SRM936a-0,08 SRM936a-0,04 X Wavelength (nm)

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U nc e r t a int y Ana lysis – Volum e

Fluore sc e nc e M e a sure m e nt s

Uncertainty Component Type Standard

Uncertainty (k=1) Relative Uncertainty Measurement repeatability A 1.0% 1.0% Wavelength B 0.10 nm 0.2% Detector linearity B <0.20% <0.20%

Radiometric calibration of emission unit

Spectral irradiance standard B 3.0 – 0.5% 3.0 – 0.5% Spectral radiance factor

standard

B 0.2% 0.2%

Transfer of relative spectral emission calibration

B 0.4% 0.4%

Total expanded (k=2) uncertainty

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Spe c t ra l Em ission Cor re c t ion

Com pa rison Re sult s

Participants:

• NIST  BAM

• PTB • NRC

U. Resch-Genger et al. Anal. Chem., 84, 3889-3898 (2012)

State-of-the-art

comparability of

corrected

emission spectra

~4.2%

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Fluore sc e nc e Re se a rch

400 500 600 700 800 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 Samples Gray Green Red Cyan R ef lec tanc e Fac tor (R ) Wavelength [nm] Colour standards: Requirement: reflectance is independent of mode of measurement

Koo, A . et. al, Appll. Opt., 49, 2375-2381(2010)

Nadal, M., Miller, C., and Zarobila, C., Color Res. Appl., (2010) Monoch. Illumination/ polych. detection

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Tra c e a bilit y of N RC Fluore sc e nc e

M e a sure m e nt s

Standard Detector 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 250 350 450 550 650 750 850 950 1050 1150 Wavelength (nm) R e sp o n si v it y s-polarized p-polarized Standard Lamp 0.0 1.0 2.0 3.0 4.0 5.0 300 350 400 450 500 550 600 650 700 750 800 Wavelength (nm) Irra d ia n ce Standard Reflector 0.99 1 1.01 1.02 1.03 350 400 450 500 550 600 650 700 750 800 850 45/ 0 R ad ian ce F ac tor

CD = calibrated detector

CS = calibrated light source

CR = calibrated reflector

CD: 1. Calibrate monitor detector CD-CR: 2. Calibrate excitation unit CS-CR: 3. Calibrate emission unit

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Spe c t ra l Em ission Ca librat ion

Three Methods:

- Physical Transfer Standards

: CS

-

CR

-

Physical Transfer Standards

: CD-

CR

- Certified Reference Material: RM

0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 350 400 450 500 550 600 650 700 750 800 Wavelength (nm) Grating B Grating A

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Ex t e nsion t o N I R:

CD

-

CR

M e t hod

)

(

)

(

)

(

)

(

2

λ

β

λ

std ref sw sys

E

k

i

R

=

0.00 0.20 0.40 0.60 300 500 700 900 1100

I

sw

(λ)

0.0 1.0 2.0 3.0 300 500 700 900 1100 Wavelength (nm)

E

ref

(

λ)

Emission Unit Spectral

Responsivity, R

sys

(

λ)

CD

Excitation Unit Spectral

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Ex t e nsion t o N I R

0.00 0.10 0.20 0.30 0.40 0.50 300 400 500 600 700 800 900 1000 1100

Responsivity Emission Unit, Grating B

R emission (Vertical) R emission (Horizontal) Wavelength (nm) 0.00 0.10 0.20 0.30 0.40 0.50 300 400 500 600 700 800 900 1000 1100

Responsivity Emission Unit Unpolarized, Xe 2011 Responsivity with Tungsten lamp (ISWB2012)

CS-CR

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Ex t e nsion t o N I R

350 440 530 0.00E+00 1.00E-05 2.00E-05 3.00E-05 4.00E-05 5.00E-05 700 750 _{800 850} 900 950 ₁₀₀₀ Excitation (nm) R aw B is p ec tr a l F a ct o r Emission (nm) 350 480 0 0.0001 0.0002 0.0003 0.0004 0.0005 700 750 800 ₈₅₀ 900 ₉₅₀ 1000 Excitation (nm) B is p ec tr a l F a ct o r Emission (nm) 360 460 560 0.00E+00 1.00E-05 2.00E-05 3.00E-05 4.00E-05 5.00E-05 Excitation (nm) Ra w B is p ec tr a l F a ct o r CCS Yellow CCS Orange

Corrected

Raw

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I m pa c t of N I R Fluore sc e nc e

0.0 0.2 0.4 0.6 0.8 1.0 1.2 300 400 500 600 700 800 900 1000 R a d ia n t P o w e r ( a .u .) Wavelength( (nm) A Equi-energy Xe source

Polychromatic illumination mode

0.0 0.2 0.4 0.6 0.8 1.0 1.2 300 400 500 600 700 800 900 1000 Radiance factor ß 45:0 Wavelength (nm)

CCS Orange

0.0 0.2 0.4 0.6 0.8 1.0 300 400 500 600 700 800 900 1000 Wavelength (nm)

CCS Yellow

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Ex t e nsion t o Ot he r Ge om e t rie s:

Gonio- a nd Sphe re

Motivation:

Research Optical Properties of Materials

 Gonio-characteristics of fluorescent white standards

New Calibration Services

 Gonio-apparent fluorescent materials(security, decorative materials)

Standardization Procedures

 Study geometric dependence: primary c.f. transfer calibrations

- Reference measurements: bidirectional geometry (45:0) c.f.

- Commercial instruments & Standard Test Methods (ISO, ASTM, AATCC) : sphere geometry (d:0, d:8).

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Goniospe c t rofluorim e t e r Fa c ilit y

Gonio – variable angles of incidence and detection

Donated to NRC in 2009 by 3M Co.

Complements the NRC Reference Spectrofluorimeter

Gonio- spectral fluorescence: Spectral range: 300- 850 nm Spectral bandpass: 5 nm Incidence: 0° to 90°

Viewing: 22° to 180°

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Seve ra l M e a sure m e nt Ge om e t rie s

Bidirectional reflectance (45a:0)

Hemispherical reflectance (sphere method)

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Goniospe c t rofluorim e t e r

Xe source + excitation mono Sample compartment Emission mono + PMT detector

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Pre lim ina r y N RC Re sult s

S/N ~ 5:1

=> Need to

Increase

S

and

Decrease

N

Increase S: - Re-coat mirrors - Refurbish predisperser - Re-optimize optical design - Improve optical alignment

Decrease N:

-Optimize lock-in operation: phase

-Optimize software :data -4E-05 0 4E-05 8E-05 0.00012 0.00016 0.0002 500 550 600 650 700

Sample: SFS-336 , 0/45, raw data

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Cur re nt N RC Re sult s

Increase S:

- Change to AR-coated quartz beamsplitter

Decrease N:

-Add order-sorting filters -Improve stability of lock-in & chopper method -2E-05 3E-05 8E-05 0.00013 0.00018 0.00023 500 550 600 650 700

Sample: SFS-336 , 0/45, EX=350 nm, raw data

Sep2010 Apr2012

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Ca librat ion of Em ission U nit –

Ca librat e d Sourc e M e t hod

Spectral radiance

Spectral reflectance standard

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M e a sure d Em ission U nit Dat a (ra w )

0 0.002 0.004 0.006 0.008 0.01 0.012 0.014 0.016 330 430 530 630 730 830 0/25 0/30 0/35 0/40 0/45 0/50 0/55 0/60 0/65 0/70 0/75 0/80 Wavelength (nm)

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Tra c e a bilit y: Spe c t ra l Em ission Dat a

Calibrated spectral reflectance standard (pressed PTFE) BRDF data: 0/25 to 0/80 degrees

Calibrated spectral radiance sphere source 0.8 0.85 0.9 0.95 1 1.05 1.1 400 500 … 600 700 Wavelength (nm) 0 100 200 300 400 500 600 700 800 900 380 580 780 980 Wavelength (nm)

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Em ission U nit Spe c t ra l

Cor re c t ion

0 1 2 3 4 5 350 450 550 650 750 25 30 35 40 45 50 55 60 65 70 75 80 Corrected for: • Gonio/spectral of PTFE reflectance standard • Spectral transmittance of ND filter (0.7) • Standard spectral radiance source Spectral Correction – NOT GEOMETRY-DEPENDENT Wavelength (nm) (R e spo ns iv it y )-1

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Cur re nt N RC Goniofluore sc e nc e

M e a sure m e nt Re sult s

0 0.00005 0.0001 0.00015 0.0002 0.00025 0.0003 500 550 600 650 700 SFS-336 , 0/45, EX=350 nm - Corrected