interpretation of vsn normalized data
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@maurice-melancon-1611
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@maurice-melancon-1611
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@wolfgang-huber-3550
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EMBL European Molecular Biology Laborat…
Hi Maurice, in statistics it is sometimes useful to differentiate between (a) the estimator and (b) the true underlying quantity that you want to estimate. For example, if you want to estimate the expectation value of a symmetric distribution, you can use the mean, or the median as estimators. They are both correct, but depending on the data they can provide different, and more or less appropriate answers. With microarrays, (b) is the fold-change, that is the change in mRNA abundance. The log-ratio of fluorescence intensities is a simple and intuitive estimator for this, but if the fluorescence intensities become small, this estimator can have unpleasant properties, like large variance. The glog-ratio (what vsn provides) is an alternative estimator, which avoids the variance explosion, for the price of being biased towards 0 when the fluorescence intensities are small. Note that the vsn function returns glog to base e (so a glog-ratio of 1 corresponds to an estimated fold change of exp(1) = 2.718..) while many other packages use log2. Hope this helps Wolfgang Maurice Melancon wrote: > Hello All, > > I used vsn to normalze my one-channel cDNA microarray experiment. I'm sorry > if this is an elementary question (I'm not a math person) but can vsn data > be interpreted in similar fashion to log2 data, e.g. 1 log vale = 2-fold > induction? What would be the appropriate transformation to get to either > log2 or raw data from vsn data? > > Briefly, what I did was to normalize using vsn, then I used SAS to run > anovas with pairwise comparisons and anova slicing. Using the estimate > function returns estimated differences between the reported means. I am > seeking then to bridge the gap between these estimates and actual fold > changes. I think this can be done, but I am unsure about how to either > reverse-transform the vsn data or how to interpret it biologically (e.g. 1 > log = 2x fold change) > > WIth thanks > > Maurice > > [[alternative HTML version deleted]] > > _______________________________________________ > Bioconductor mailing list > Bioconductor at stat.math.ethz.ch > https://stat.ethz.ch/mailman/listinfo/bioconductor -- Best regards Wolfgang ------------------------------------- Wolfgang Huber European Bioinformatics Institute European Molecular Biology Laboratory Cambridge CB10 1SD England Phone: +44 1223 494642 Fax: +44 1223 494486 Http: www.ebi.ac.uk/huber
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I agree. There is a difference between hypothesis testing and estimation. Microarray is good for hypothesis testing (and hypothesis generating) but not good at estimation. Consider the following two observations : 1) Different platforms and softwares (e.g. preprocessing algorithm) give rise to different dynamic ranges and there is no agreed gold standard. 2) It is common that biologist validate the genes ranked highly by microarray results with a method such as real time PCR to estimate the fold change accurately. Given these two observations I do not see why biologists are obsessed by the need to observe a two-fold change by microarrays ! Therefore whether the log is to the base 2, 10 or natural is irrelevant. Regards, Adai On Thu, 2006-02-16 at 18:16 +0000, Wolfgang Huber wrote: > Hi Maurice, > > in statistics it is sometimes useful to differentiate between (a) the > estimator and (b) the true underlying quantity that you want to estimate. > > For example, if you want to estimate the expectation value of a > symmetric distribution, you can use the mean, or the median as > estimators. They are both correct, but depending on the data they can > provide different, and more or less appropriate answers. > > With microarrays, (b) is the fold-change, that is the change in mRNA > abundance. The log-ratio of fluorescence intensities is a simple and > intuitive estimator for this, but if the fluorescence intensities become > small, this estimator can have unpleasant properties, like large > variance. The glog-ratio (what vsn provides) is an alternative > estimator, which avoids the variance explosion, for the price of being > biased towards 0 when the fluorescence intensities are small. > > Note that the vsn function returns glog to base e (so a glog-ratio of 1 > corresponds to an estimated fold change of exp(1) = 2.718..) while many > other packages use log2. > > Hope this helps > Wolfgang > > > > > Maurice Melancon wrote: > > Hello All, > > > > I used vsn to normalze my one-channel cDNA microarray experiment. I'm sorry > > if this is an elementary question (I'm not a math person) but can vsn data > > be interpreted in similar fashion to log2 data, e.g. 1 log vale = 2-fold > > induction? What would be the appropriate transformation to get to either > > log2 or raw data from vsn data? > > > > Briefly, what I did was to normalize using vsn, then I used SAS to run > > anovas with pairwise comparisons and anova slicing. Using the estimate > > function returns estimated differences between the reported means. I am > > seeking then to bridge the gap between these estimates and actual fold > > changes. I think this can be done, but I am unsure about how to either > > reverse-transform the vsn data or how to interpret it biologically (e.g. 1 > > log = 2x fold change) > > > > WIth thanks > > > > Maurice > > > > [[alternative HTML version deleted]] > > > > _______________________________________________ > > Bioconductor mailing list > > Bioconductor at stat.math.ethz.ch > > https://stat.ethz.ch/mailman/listinfo/bioconductor > >
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