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Repeatability – Pooling Variances Stanley N. Deming, Ph.D. Statistical Designs 9941 Rowlett Road, Suite 10 Houston, TX 77075-3404 U.S.A. 713 947 1551 [email protected] BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 1 Pooling: Fundamental Definition Of Variance An estimated variance, s2, is a sum of squares (SS) divided by a degrees of freedom (df): sum of squares s variance degrees of freedom For example, the familiar correct equation for the estimated standard deviation of n data points about their mean is n 2 An estimated standard deviation, s, is the square root of an estimated variance, s2: s ( xi x )2 i 1 n 1 where the numerator is n s s2 variance SS df ( xi x )2 sum of squared deviations i 1 and the denominator is n 1 degrees of freedom of deviations Because one degree of freedom has been used to calculate the mean, only n - 1 of the deviations are independent. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 2 Pooling: A Measurement Method As A System A measurement method may be represented with a systems theory diagram. The "Measurement Measurement Method" Method converts information contained in the sample into numerical results. The "Sample" Sample (the input) is usually a physical material to be characterized. The "Results" (the output) are a set of data. For our purposes here, the "measurement method" may be defined as a bioassay. A caution about words: Statisticians call the results a "statistical sample," a subset of a few of the conceptually infinite number of individual results that could have been obtained from the "physical sample." The physical sample on the left in the figure is not the same as the statistical sample on the right in the figure. (Don’t use the word " "sample" l " iin a conversation i with iha statistician without putting an adjective in front of it!) BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 3 Pooling: Common Measurement Practice Many measurement laboratories make multiple measurements on each sample that is submitted. These are called replicate measurements — they are repeated measures of the same thing. The figure at the right shows three replicate measurement results (the three circles at the right, n = 3) for a single sample of material. Many measurement courses (undergraduate qua t tat e a quantitative analysis, a ys s, for o e example) a p e) po pointt out that it requires at least two replicate measurements to calculate a standard deviation, and taking three measurements on each p physical y sample p is a common p practice. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 4 Pooling: The Mean And Standard Deviation For One Sample Suppose the three replicate measurements give the values 61.5, 63.3, and 62.7. These three values are graphed at the bottom right. The vertical dimension has no meaning here — it is just a convenient place to put the data points. The average is x 61.5 63.3 62.7 62.50 3 and the standard deviation s is 3 ( xi 62.50)2 i 1 s 3 1 0.9165 This estimate of has only 2 degrees of freedom. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 5 Pooling: The Mean And Standard Deviation For One Sample In the graph at the bottom right, some lines have been added. The vertical line is drawn at 62.50, the mean of the three data points. For each data point, a horizontal line has been drawn from that data point to the mean. These horizontal lines represent the residuals that are squared and summed to calculate the standard sta da d de deviation. at o The results are reported to the client: mean std. dev. n = = = 62.50 0.9165 3 ((Reporting p g 62.50 0.92 is discouraged.) g ) So much for the first sample. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 6 Pooling: A Second Sample The figure at the right shows three replicate measurement results for a second physical sample of material (three more circles at the right). BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 7 Pooling: A Second Sample Suppose this second sample gives measured values of 66.9, 67.2, and 66.3. These three values are graphed at the bottom right, along with the data from the first sample. Again, the vertical dimension has no meaning. The average is x 66.9 67.2 66.3 66.80 3 and the standard deviation s is 3 s ( xi 66.80)2 i 1 3 1 0.4583 This estimate of has only 2 degrees of freedom. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 8 Pooling: A Second Sample In the graph at the bottom right, some lines have been added. The vertical line is drawn at 66.80, the mean of the three data points. For each data point, a horizontal line has been drawn from that data point to the mean. These horizontal lines represent the residuals that are squared and summed to calculate the standard sta da d de deviation. at o The results are reported to the client: mean std. dev. n = = = 66.80 0.4583 3 So much for the second sample. p BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 9 Pooling: The Source Of Variation For The Two Samples The two means (62.50 and 66.80) differ from each other because of the different characteristics of the two physical samples. Both sets of results came from the same measurement method. If the samples are homogeneous, any variation of the individual results around their separate means must be caused by the measurement method. The standard deviation is a characteristic of the o t e measurement easu e e t method, et od, not ot the t e sample! Using a new subscript to indicate the physical sample p number,, we would therefore expect p the ( x1i x1) deviations to be similar to the ( x2i x2 ) deviations. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 10 Pooling: The Source Of Variation For The Two Samples Further, the two sums of squares n1 ( x1i x1)2 i 1 and n2 ( x2i x2 )2 i 1 should be two separate p estimates of the variability of the measurement method. One estimate has n1 - 1 df, and the other estimate has n2 - 1 df. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 11 Pooling: The Pooled Standard Deviation Using all of the information from the two samples, the composite (or pooled) sum of squared deviations from the means is n1 n2 i 1 i 1 ( x1i x1)2 ( x2i x2 )2 and the composite p ((or p pooled) degrees g of freedom is (n1 - 1) + (n2 - 1) = n1 + n2 – 2 Returning R t i tto th the ffundamental d t ld definition fi iti off variance, and substituting the pooled sum of squares and pooled degrees of freedom gives n1 sp n2 ( x1i x1)2 ( x2i x2 )2 i 1 i 1 n1 n2 2 Because it has more degrees of freedom, sp is a better estimate of . BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 12 Pooling: Generalizing To Many Statistical Samples If j is an index that refers to a particular statistical sample, then the notation 2 j=1 indicates a summation over two statistical samples. The equation for the pooled standard deviation n1 sp n2 ( x1i x1)2 ( x2i x2 )2 i 1 i 1 n1 n2 2 can be rewritten 2 nj sp ( x ji x j )2 j 1 i 1 2 nj 2 j 1 BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 13 Pooling: The Method Standard Deviation, m In general, if J represents the total number of statistical samples, then J m sp nj ( x ji x j )2 j 1 i 1 J nj J j 1 Many laboratories make multiple measurements on each physical sample that is analyzed (see the figure at the right). This historical information can be used to calculate a pooled standard deviation. Because the number of degrees of freedom is so large (e.g., 19,054), sp becomes a very good estimate of for the measurement method. This population parameter is given the subscript "m" m and is called a "method method standard deviation": m = 0.8159, for example. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 14 Pooling: An Alternative Protocol For Measurement When m is known, it isn't necessary to make multiple measurements. Future samples can be measured only once. In the past, measurements were reported using the mean and the standard deviation calculated for each sample: x1 62.50 s1 0.9165 n3 x2 66.80 s2 0.4583 n3 In the future, measurements can be reported using a mean of only one measurement and the method standard deviation x9,528 63.74 x9,529 58.67 m 0.8159 m 0.8159 n 1 n 1 This saves the cost of extra measurements and gives the customer a consistent (and more realistic) estimate of the uncertainty. (Scared to do this? Think again. When you have your cholesterol measured, how many measurements does your health care plan pay for? They know their m.) BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 15 Pooling: An Alternative Protocol For Measurement The confidence interval of the mean does decrease with multiple measurements: x z m n When n = 3, the confidence interval is about 42% narrower than when n = 1: tripling the cost is a big price to pay for a slight narrowing in the confidence interval. If the data are proportionally heteroscedastic, a a an analogous a ogous app approximate o ate equat equation o ca can be used to pool the relative standard deviation: 2 x ji x j x j m j 1 i 1 J nj J j 1 However, if the measured values are small and have a homoscedastic component, this equation will give an inflated estimate. J nj BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 16 Pooling: An Alternative Calculation When the standard deviations of the individual samples have already been calculated, an alternative equation for calculating the pooled standard deviation can be used: sp (n1 1)s12 (n2 1)s22 n1 n2 2 Recall that an estimated variance is a sum of squares divided by its degrees of freedom. Multiplying an estimated variance by its degrees of freedom will ill "regenerate" the ssum m of squares. For example, n1 s12 ( x1i x1)2 i 1 (n1 1) ; (n1 1)s12 n1 ( x1i x1)2 i 1 If the reported values of the standard deviation have been rounded (e (e.g., g 0 0.5 5 instead of 0.4582575...), this alternative calculation might give distorted results. BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 17 Pooling: Where Does The Variance Come From? The "measurement method" might be larger than has been assumed up to this point. In bioassays, it might be more than just the plate on which reagents have been added and dilutions have been made. Just what is meant by "the measurement method depends on the definition of the method" boundaries of the "measurement system." Thus hairs are split about "repeatability," "intermediate te ed ate p precision," ec s o , etc. etc Nonetheless, once the "measurement method" and its sources of variation has been defined,, the resulting g replicate p data can be pooled as shown in this presentation. (Nested, split-plot, or staggered-nested experimental p designs g can be used to estimate individual sources of variation. But that’s another story.) BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 18 Repeatability – Pooling Variances (END) Stanley N. Deming, Ph.D. Statistical Designs 9941 Rowlett Road, Suite 10 Houston, TX 77075-3404 U.S.A. 713 947 1551 [email protected] BEBPA Nice 28 September 2011. DO NOT COPY Copyright © 2011 by Stanley N. Deming. All rights reserved. Slide 19