In LeiLi-Uchicago/Cookie: An sampling method to select representative samples from large population

knitr::opts_chunk$set(echo = TRUE, cache = FALSE)

Prerequisite

if(!require(devtools)){
  install.packages("devtools", quiet = TRUE)
}

if(!require(ggradar)){
  devtools::install_github("ricardo-bion/ggradar", quiet = TRUE)
}

Load package

library(Cookie)
library(cowplot)
library(ggplot2)

Load data

User can use read.csv() to load data into R. Columns should be factors, and each sample should have an unique name or ID (in this example is cell barcode). The missing values should be indicated by "NA" in CSV file. Users can arrange and edit their data in Excel and then import into R.

data <- read.csv(file = system.file("extdata", "data.csv", package = "Cookie"), header = TRUE, row.names = 1)

Create object

Create data object and determine factor types

cookie <- createCookieObject(data=data,factor.type = c("char","char","char","char","num"))

Normalize data (for numerical factors)

cookie <- normalization(cookie)

Calculate pair-wise distances

cookie <- distCalculation(cookie)

Generate non-linear dim reduction

Users also can use UMAP (https://github.com/tkonopka/umap) to generate non-linear reduction. Two methods is available: "naive" and "umap-learn". To use "umap-learn", users may need to install python package "umap-learn" (https://pypi.org/project/umap-learn/).

cookie <- reductionTSNE(cookie)
#cookie <- reductionUMAP(cookie)

See how factors distributed

p1 <- plotReduction(cookie, reduction = "tsne", group.factor = "Subject") 
p2 <- plotReduction(cookie, reduction = "tsne", group.factor = "Isotype") 
plot_grid(p1,p2,labels = c("Subject","Isotype"), ncol = 2)

Test coverages for different sample size to determine an appropriate sample size

we run two test 1) with prime factor 2) without prime factor

cookie <- sampleSizeTest(cookie, prime.factor = "Subject",size.range = c(3,5,7,9,11,13,15),name = "run1")

# When selecting samples from entire large population, users can use fast.mode = 3, 4, or 5 to enable FastPAM algorithm.  
cookie <- sampleSizeTest(cookie, size.range = c(60,70,80,90,100,110,120),name = "run2", fast.mode = 5)

1) with prime factor, coverage rate of each sample size

p1 <- plotSizeTest(cookie, test.name = "run1", chart.type = "line", coverage.type = "pct")
p2 <- plotSizeTest(cookie, test.name = "run1", chart.type = "radar", coverage.type = "pct")

plot_grid(p1,p2,ncol = 2, rel_widths = c(1,1.5))

2) without prime factor, coverage rate of each sample size

p1 <- plotSizeTest(cookie, test.name = "run2", chart.type = "line", coverage.type = "pct")
p2 <- plotSizeTest(cookie, test.name = "run2", chart.type = "radar", coverage.type = "pct")

plot_grid(p1,p2,ncol = 2, rel_widths = c(1,1.5))

After determine an appropriate sample size, users can select samples from original population

we run two sampling 1) with prime factor 2) without prime factor

cookie <- sampling(cookie, prime.factor = "Subject",important.factor = c("Isotype"),sample.size = 5,name = "run1")

# When selecting samples from entire large population, users can use fast.mode = 3, 4, or 5 to enable FastPAM algorithm.(https://stat.ethz.ch/R-manual/R-patched/library/cluster/html/pam.html)
cookie <- sampling(cookie, important.factor = c("Isotype","Subject"),sample.size = 90, name = "run2", fast.mode = 5)

Coverage on each factor of current sampling 1) with prime factor Because all samples were selected evenly from each subject, the STD of selected samples on subject is 0. The CC value is not available. We set CC to 0.

p1 <- plotSampling(cookie,name = "run1", coverage.type = "pct") + theme(legend.position = "none")
p2 <- plotSampling(cookie,name = "run1", coverage.type = "cc") + theme(legend.position = "none")

plot_grid(p1,p2,labels = c("Coverage rates","CC values between original and selection"), ncol = 2)

Coverage on each factor of current sampling 2) without prime factor

p1 <- plotSampling(cookie,name = "run2", coverage.type = "pct") + theme(legend.position = "none")
p2 <- plotSampling(cookie,name = "run2", coverage.type = "cc") + theme(legend.position = "none")

plot_grid(p1,p2,labels = c("Coverage rates","CC values between original and selection"), ncol = 2)

Selected samples loacted on 2D map

p1 <- plotReduction(cookie, reduction = "tsne", group.factor = "Subject", sampling = "run1") 
p2 <- plotReduction(cookie, reduction = "tsne", group.factor = "Isotype", sampling = "run1") 
plot_grid(p1,p2,labels = c("Subject","Isotype"), ncol = 2)

Distribution of original population and current sampling

p1 <- plotDistribution(cookie, sampling = "original",factor = "Subject") + labs(title = "")
p2 <- plotDistribution(cookie, sampling = "original",factor = "Seuret_Cluster") + labs(title = "")
p3 <- plotDistribution(cookie, sampling = "original",factor = "Isotype") + labs(title = "")
p4 <- plotDistribution(cookie, sampling = "original",factor = "V.locus") + labs(title = "")
p5 <- plotDistribution(cookie, sampling = "original",factor = "CDR3.length") + labs(title = "")

p6 <- plotDistribution(cookie, sampling = "run2",factor = "Subject") + labs(title = "")
p7 <- plotDistribution(cookie, sampling = "run2",factor = "Seuret_Cluster") + labs(title = "")
p8 <- plotDistribution(cookie, sampling = "run2",factor = "Isotype") + labs(title = "")
p9 <- plotDistribution(cookie, sampling = "run2",factor = "V.locus") + labs(title = "")
p10 <- plotDistribution(cookie, sampling = "run2",factor = "CDR3.length") + labs(title = "")

plot_grid(p1,p2,p3,p4,p5,p6,p7,p8,p9,p10, ncol = 5, labels = c("Original-Subject","Original-Cluster","Original-Isotype","Original-V.locus","Original-CDR3.length","Sampling-Subject","Sampling-Cluster","Sampling-Isotype","Sampling-V.locus","Sampling-CDR3.length"))

LeiLi-Uchicago/Cookie documentation built on Jan. 26, 2024, 2:01 p.m.