First install the auxilliary packages for conos, conosPanel:
install.packages('conosPanel', repos='https://kharchenkolab.github.io/drat/', type='source')Now load the conos library, and the R package conosPanel for the example data panel:
library(conos)
panel <- conosPanel::panelNext, use pagoda2 for pre-processing:
library(pagoda2)
panel.preprocessed <- lapply(panel, basicP2proc, n.cores=1, min.cells.per.gene=0, n.odgenes=2e3,
get.largevis=FALSE, make.geneknn=FALSE)## creating space of type angular done
## adding data ... done
## building index ... done
## querying ... done
## creating space of type angular done
## adding data ... done
## building index ... done
## querying ... done
## creating space of type angular done
## adding data ... done
## building index ... done
## querying ... done
## creating space of type angular done
## adding data ... done
## building index ... done
## querying ... done
Now align the datasets:
con <- Conos$new(panel.preprocessed, n.cores=1)
con$buildGraph(k=15, k.self=5, space='PCA', ncomps=30)## .............
Next find the clusters, and create an embedding:
con$findCommunities()
con$embedGraph(method="UMAP")## Estimating hitting distances: 05:07:28.
## Done.
## Estimating commute distances: 05:08:20.
## Hashing adjacency list: 05:08:20.
## Done.
## Estimating distances: 05:08:23.
## Done
## Done.
## All done!: 05:08:29.
Now prepare the metadata (which can be any type of clustering of all the cells):
metadata <- data.frame(Cluster=con$clusters$leiden$groups)Save data (set exchange_dir to your path):
## use current directory
exchange_dir <- "."
hdf5file = "example.h5"
saveConosForScanPy(con, output.path=exchange_dir, hdf5_filename=hdf5file, verbose=TRUE)Users can then access the data saved to the HDF5 file, e.g. to access metadata, run:
library(rhdf5)
metadata = h5read(paste0(exchange_dir, "/example.h5"), 'metadata/metadata.df')
head(metadata, 4)## CellId Dataset
## 1 MantonBM1_HiSeq_1-TCTATTGGTCTCTCGT-1 MantonBM1_HiSeq_1
## 2 MantonBM1_HiSeq_1-GAATAAGTCACGCATA-1 MantonBM1_HiSeq_1
## 3 MantonBM1_HiSeq_1-ACACCGGTCTAACTTC-1 MantonBM1_HiSeq_1
## 4 MantonBM1_HiSeq_1-TCATTTGGTACGCTGC-1 MantonBM1_HiSeq_1
All possible fields included in the output HDF5 file are:
raw_count_matrix: the sparsedgCMatrixof raw countsraw_count_matrix/data: the matrix entriesraw_count_matrix/shape: the matrix dimensionsraw_count_matrix/indices: 0-based vector of non-zero matrix entriesraw_count_matrix/indptr: vector of pointers, one for each column (or row), to the initial (zero-based) index of elements in the column (or row)
metadata/metadata.df: thedata.frameof metadata valuesgenes/genes.df: thedata.frameof genescount_matrix: the sparsedgCMatrixof normalized counts, ifcm.norm=TRUEcount_matrix/data: the matrix entriescount_matrix/shape: the matrix dimensionscount_matrix/indices: 0-based vector of non-zero matrix entriescount_matrix/indptr: vector of pointers, one for each column (or row), to the initial (zero-based) index of elements in the column (or row)
embedding/embedding.df: thedata.frameof the conos embedding, ifembedding=TRUEpseudopca/pseudopca.df: thedata.frameof an emulated PCA by embedding the graph to a space withn.dimsdimensions and save it as a pseudoPCA, ifpseudopca=TRUEpca/pca.df: thedata.frameof the PCA of all the samples (not batch corrected), ifpca=TRUEgraph_connectivities: the sparsedgCMatrixof graph connectivites, ifalignment.graph=TRUEgraph_connectivities/data: the matrix entriesgraph_connectivities/shape: the matrix dimensionsgraph_connectivities/indices: 0-based vector of non-zero matrix entriesgraph_connectivities/indptr: vector of pointers, one for each column (or row), to the initial (zero-based) index of elements in the column (or row)
graph_distances: the sparsedgCMatrixof graph distances, ifalignment.graph=TRUEgraph_distances/data: the matrix entriesgraph_distances/shape: the matrix dimensionsgraph_distances/indices: 0-based vector of non-zero matrix entriesgraph_distances/indptr: vector of pointers, one for each column (or row), to the initial (zero-based) index of elements in the column (or row)
In order to read in the dcGMatrix again, simply use the Matrix package as follows:
library(rhdf5)
library(Matrix)
rawcountMat = h5read(paste0(exchange_dir, "/example.h5"), 'raw_count_matrix')
raw_count_matrix = sparseMatrix(x = as.numeric(rawcountMat$data),
dims = as.numeric(c(rawcountMat$shape[1], rawcountMat$shape[2])),
p = as.numeric(rawcountMat$indptr),
i = rawcountMat$indices, index1=FALSE)Note: Please set index1=FALSE as the index vectors are 0-based. For more details, see the documentation for Matrix::sparseMatrix()