olin {OLIN} | R Documentation |
This functions performs optimised local intensity-dependent normalisation (OLIN) and optimised scaled intensity-dependent normalisation (OSLIN).
olin(object,X=NA,Y=NA,alpha=seq(0.1,1,0.1),iter=3, scaling=FALSE,scale=c(0.05,0.1,0.5,1,2,10,20),weights=NA)
object |
object of class “marrayRaw” |
X |
matrix with x-coordinates of spots. If X=NA, columns on array are used as proxies for the location in x-direction |
Y |
matrix with y-coordinates of spots. If Y=NA, rows on array are used as proxies for the location in y-direction |
alpha |
vector of alpha parameters that are tested in the GCV procedure |
iter |
number of iterations in the OLIN procedure |
scaling |
If scaling=TRUE, a subsequent optimised scaling is performed |
scale |
vector of scale parameters that are tested in a GCV procedure. |
weights |
matrix of weights for local regression. Rows correspond to the spotted probe sequences, columns to arrays in the batch. These may be derived from the matrix of spot quality weights as defined for “maRaw” objects. |
OLIN and OSLIN are based on iterative local regression and incorporate optimisation of model parameters.
Local regression is performed using LOCFIT, which requires the user to choose a specific smoothing parameter alpha
that controls the neighbourhood size h of local fitting. The parameter alpha
specifies the fraction of points that are included in the neighbourhood and thus has a value between 0 and 1.
Larger alpha
values lead to smoother fits.
Additionally, the setting of scale parameters controls for distinct amount of smoothing in
Y-direction compared to smoothing in X-direction.
The parameter scale
can be of arbitrary value.
The choice of model parameters alpha
and scale
for local regression is crucial for the efficiency and
quality of normalization. To optimize the model parameters, a general cross-validation procedure (GCV) is applied.
The arguments alpha
and scale
define the parameters values which are tested in the GCV.
Detailed information about OLIN and OSLIN can be found in the package documentation and in the reference stated below.
The weights argument specifies the influence of the single spots on the local regression. To exclude
spots being used for the local regression (such as control spots), set their corresponding weight to zero.
Note that OLIN and OSLIN
are based on the assumptions that most genes are not differentially expressed (or up- and down-regulation
is balanced) and that genes are randomly spotted across the array. If these assumptions are not valid, local
regression can lead to an underestimation of differential expression. OSLIN is especially sensitive to violations of these assumptions. However, this
sensitivity can be decreased if the minimal alpha
-value is increased. Minimal alpha
defines the
smallest scale used for local regression. Increasing alpha
can reduce the influence of localised
artifacts as a larger fraction of data points is included.
It is also important to note that OLIN/OSLIN is fairly efficient in removing intensity- and spatial-dependent dye bias, so that normalised data will look quite “good” after normalisation independently of the true underlying data quality. Normalisation by local regression assumes smoothness of bias. Therefore, localised artifacts such as scratches, edge effects or bubbles should be avoided. Spots of these areas should be flagged (before normalisation is applied) to ensure data integrity. To stringently detect artifacts, the OLIN functions fdr.int, fdr.spatial, p.int
and p.spatial
can be used.
Object of class “marrayNorm” with normalised logged ratios
Matthias E. Futschik (http://itb.biologie.hu-berlin.de/~futschik)
# LOADING DATA data(sw) data(sw.xy) # OPTIMISED LOCAL INTENSITY-DEPENDENT NORMALISATION OF FIRST ARRAY norm.olin <- olin(sw[,1],X=sw.xy$X[,1],Y=sw.xy$Y[,1]) # MA-PLOT OF NORMALISATION RESULTS OF FIRST ARRAY plot(maA(norm.olin),maM(norm.olin),main="OLIN") # CORRESPONDING MXY-PLOT Mtmp <- mxy.plot(maM(norm.olin)[,1],Ngc=maNgc(norm.olin),Ngr=maNgr(norm.olin), Nsc=maNsc(norm.olin),Nsr=maNsr(norm.olin),main="OLIN") # OPTIMISED SCALED LOCAL INTENSITY-DEPENDENT NORMALISATION norm.oslin <- olin(sw[,1],X=sw.xy$X[,1],Y=sw.xy$Y[,1],scaling=TRUE) # MA-PLOT plot(maA(norm.oslin),maM(norm.oslin),main="OSLIN") # MXY-PLOT Mtmp <- mxy.plot(maM(norm.oslin)[,1],Ngc=maNgc(norm.oslin),Ngr=maNgr(norm.oslin), Nsc=maNsc(norm.oslin),Nsr=maNsr(norm.oslin),main="OSLIN")