Narrow regions exhibiting a high point density are mapped to clusters; their centroid is taken as the 3D position of a marker and used as an input to determine the projection traces.
Despite the misshapen projections, we were able to determine the projection direction for these mutants (Supporting Information, Fig. S4).
To determine the projection site of GRP-immunoreactive axons, double-immunofluorescence staining of GRP (1∶5,000 dilution) and nNOS (A-11; mouse monoclonal antibody, Santa Cruz Biotechnology, Santa Cruz, CA) (1∶8,000 dilution), a marker protein for neurons in the SPN, was performed as described previously [20].
It would be interesting to determine the projection(s) of these 2 types of neurons.
In a next step, optical variables can be computed by determining the projection of the reconstructed physical ball trajectory onto an image plane.
LDA is the classifier that separates the two or more classes by determining the projection matrix that maximizes the ratio of between-class covariance to within-class covariance [ 25].
The two terms empirically determine the projection quality by quantifying for each MCEP, how wide its n most similar MCEPs are scattered across the grid (C n ) and how many non-similar, i.e. false MCEPs have been wrongly mapped into the vicinity in the grid (see the Supplementary Material B for details please).
For further identification of theses neuroblast lineages, we determined the projection patterns of each of their secondary axon tracts relative to the ensemble of secondary axon tracts in the late larval brain based on anti-NRT immunolabeling and compared these patterns to those documented in previous lineage mapping studies (Pereanu and Hartenstein, 2006; Spindler and Hartenstein, 2010).
However, in a mix-and-match experiment, in which the BNST of males (or testosterone-treated females) was co-cultured with the AVPV of females (or vice versa), it was clear that hormone action at the target site (AVPV) determines the size of the projection from the BNST [ 41].
The present study was designed to determine the differential projection pattern as well as the laterality of descending TMN projections to the PPTg and LDTg.
