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Calibration and leak dataA. Neilson

June 27, 2008

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Curve data

Curve data 0608

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0 5000 10000 15000 20000 25000 30000 35000

CNT (normalized to 10K amb)

PP

O%

Buck measurement

Avg SH 6-08

Power fit inst

Ksv .0143

Superimposed data sets from SH and Buck measurements over Curve Ksv (K=.0143), and Curve (9.16e19X^-4.44)

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Leak through wall

24 wellAmbient dO2 in H2O 0.00685 gmCO2/LWt of O2 32 gm/moleStarting concentration 2.14E-04 mol/literH2O density 1000 gm/literWt of H2O 18 gm/molH2O concentration 5.56E+01 mol/literDelta O2% 21.0000% Differential O2 @ 0 PPO insideDO content of H2O 0.007 gmO2/L O2 content of water DO/% 0.033333333Diameter 6.00E+00 mm Diameter of wellArea bottom 28.27 mm^2 Bottom of wellHeight 0.24 mm Height of measurement chamberArea wall 4.52E+00Area total 3.28E+01 mm^2 Total area around small volumeWall thickness 0.50 mmWell volume 6.79E-06Conversion 1000.00 mm/mConversion 60.00 sec/minPET permeability 7.20E-11 gm/m.s Solubility coeff (grm/M^3)* diffusion coef(mm2/sec)PS permeability 2.40E-09 gm/m.s

Theoretical max Leak through wall PET 1.86 pmol/min Theoretical outagassing based on permeability coefTheoretical max leak through wall PS 61.99 pmol/min Theoretical outagassing based on permeability coef

Leakage through wall of plate

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Leak around perimeter of probe

Well diameter Dia 6.00E+00 mmArea A 28.27 mm^2Height h 5.00 mm Distance from top of media to chamber in plunged positionProbe diameter 5.55 mmProbe Area 24.19 mm^2Open area 4.08 mm^2

Diffusion constant D 2.00E-03 mm^2/sec Fick's constant

Theoretical Max O2 diffusion 21.43 pmoles/minmaximum based on zero O2 at cells OCR=D*S*l*dP/h (cells are in a monolayer at the bottom of well with zero O2)

DO content of H2O 0.007 gmO2/L O2 content of water Mole O2 weighs 32 gm/mole From periodic chartPP O2 152 mmHg partial pressure of O2 at 37C, 1atmMol/l O2 2.188E-04 mol/l Delta O2 concentration (air approx @37C, 152mm PPO)

Leakage around perimter of probe

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Leak data

Total leak

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%O2

Lea

k ra

te p

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l/min

Total leak

Total leak is the theoretical leakage as a result of both perimeter leak and wall leak. Both are more significant than originally thought! %O2 id the dP across the boundary condition.

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2008-06-11 Curve

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Leak back

Solubility (@ 6.79ul)= 9.6pmol/mm

Back leakage = 10pmol/min @ dP=35mm (dP=20-5=15%)

Based on theoretical leak it should be roughly 60pmol/min

Difference between theoretical and actual is likely due to the complex geometry and actual path length to the sensor

Leak rate 1mm/min

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2008-06-11 Ksv

Why do these rates seem so slow? 10ugms should be 100pmol/min basal and 5-10X with FCCP, 1000pmol/min??