QC for SDY 820

library(flowCore)
library(flowWorkspace)
# library(cytoqc)
devtools::load_all()

Load data

path <- "~/remote/fh/fast/gottardo_r/mike_working/flowcap3/"
files <- list.files(path, pattern = ".fcs", full.names = T)
set.seed(1)
files <- sample(files, 40)
cqc_data <- cqc_load_fcs(files, which.lines = 1)

Check channel

check_res <- cqc_check(cqc_data, "channel")
check_res
group_id nFCS channel
1 40 B515-A, B710-A, FSC-A, FSC-H, G560-A, G610-A, G660-A, G710-A, G780-A, R660-A, R710-A, R780-A, SSC-A, Time, V450-A, V545-A, V565-A, V585-A, V605-A, V655-A, V705-A, V800-A

Check marker

check_res <- cqc_check(cqc_data, "marker")
check_res
group_id nFCS marker
1 15
4 9 CCR7, CD107A, CD154, CD27, CD3, CD4, CD45RO, CD57, CD8, IFNG, IL2, TNFA, VIVID/CD14
2 6 107a, 154, 45ro, ccr7, cd27, cd3, cd4, cd57, cd8, ifng, il2, tnfa, vivid/14
3 4 107a, ccr7, cd154, cd27, cd3, cd4, cd45ro, cd57, cd8, ifng, il2, tnfa, vivid/14
6 4 CCR7, CD154, CD27, CD3, CD4, CD45RO, CD57, CD8, IFNG, IL2, TNFA, VIVIDCD14
5 2 ccr7, cd107a, cd154, cd27, cd4, cd45ro, cd57, cd8, d3, ifng, il2, tnfa, vivid/cd14

Match reference

match_res <- cqc_match(check_res, ref = 4, max.dist = 0.5)
match_res

Apply the match

cqc_fix(match_res)
cqc_check(cqc_data, "marker")
group_id nFCS marker
2 21 CCR7, CD107A, CD154, CD27, CD3, CD4, CD45RO, CD57, CD8, IFNG, IL2, TNFA, VIVID/CD14
1 15
3 4 CCR7, CD154, CD27, CD3, CD4, CD45RO, CD57, CD8, IFNG, IL2, TNFA, VIVID/CD14

Check panel

res <- cqc_check(cqc_data, "panel")
res

Use group 1 to standardize the rest

cqc_fix_panel(res, 2, by = "channel")
cqc_check(cqc_data, "panel")

check keywords

res <- cqc_check(cqc_data, "keyword")
res
group_id nFCS keyword
1 24 CYTNUM, EXPERIMENT NAME, FILENAME, GUID, ORIGINALGUID, PATIENT ID, PLATE ID, PLATE NAME, SAMPLE ID, transformation, TUBE NAME, WELL ID
2 16 CYTNUM, EXPERIMENT NAME, FILENAME, GUID, ORIGINALGUID, PLATE ID, PLATE NAME, transformation, TUBE NAME, WELL ID
res <- cqc_match(res, ref = 1)
res

Merge the standarized data

cs <- cytoset(cqc_data)
cs
## A cytoset with 40 samples.
## 
##   column names:
##     FSC-A, FSC-H, SSC-A, B710-A, B515-A, G780-A, G710-A, G660-A, G610-A, G560-A, R780-A, R710-A, R660-A, V800-A, V705-A, V655-A, V605-A, V585-A, V565-A, V545-A, V450-A, Time