Introduction to quantMSImageR

Introduction

A targeted DESI-MRM imaging run produces one intensity per transition per pixel, across tens of thousands of pixels and several acquisitions. Turning that into a biological result runs into three problems at once.

Most pixels are not tissue. A section occupies part of the scanned area, and the surrounding slide still returns signal – chemical background, carryover and detector noise. Left in, those pixels drag down every summary statistic and make images look more uniform than they are. Which pixels are tissue has to be decided per acquisition, and the decision has to survive being re-applied to a different MRM panel of the same sample.

Not every monitored transition yields usable analyte signal. A transition can be listed in the method and still carry no usable signal in a given section, or sit so close to background that it is indistinguishable from it. Reporting those alongside genuine measurements is how spurious hits reach a figure.

Response is not amount. Response depends on ionisation efficiency and local matrix suppression, so raw responses should not be interpreted as analyte amount, and their comparability between analytes, sections or instruments should not be assumed without appropriate normalisation and calibration.

What this package does

quantMSImageR addresses each of those: signal-to-noise (SNR) filtering against background pixels, tissue/background separation, per-feature ion images including .txt image outputs for overlaying with other imaging (H&E), quantile heatmaps – and quantification against calibration standards for the third.

Scope

The package is currently validated for Waters DESI-MRM data. The software is extendable to targeted imaging data may be analysed after conversion to a compatible MSImagingExperiment, but those workflows have not yet been formally validated.

Within that scope it is designed to handle:

  • background-referenced filtering against non-tissue pixels;
  • targeted ion-image visualisation and .txt export;
  • cross-sample summaries such as quantile heatmaps;
  • calibration-based amount estimation.

It does not do the following, by design – other tools do them better:

Not provided Use instead
Untargeted peak detection a general MSI processing package
Feature annotation / molecular identification dedicated annotation tools
Vendor-neutral raw-data conversion vendor or community converters
Histological registration TissUUmaps; our own helper tools are still in development (PMID 42438167)
General-purpose image segmentation image-analysis packages
Complex spatial / colocalisation analysis corrMSI

The workflow

Functions are grouped into families, and each section below opens with the family it belongs to. The whole workflow is:

  .raw acquisition            ion library CSV
         |                          |
         +--------> read_mrm() <----+
                        |  MSImagingExperiment, fData joined to library
                        v
        select_tissue_pixels() --> tissue_pixels.csv  (x, y, tissue/background)
                        |
    +-------------------+-------------------+
    |  ASSEMBLE   align_features -> combine_MSIs / bind_panels   |
    |             trim_MSI                                       |
    +-------------------+-------------------+
                        v   quant_MSImagingExperiment
    +-------------------+-------------------+
    |  FILTER     int2response  (IS-normalise)   |
    |             zero2na, remove_blank_mzs      |
    |             int2snr -> applySNR, back2NA   |
    +-------------------+-------------------+
                        v
         +--------------+--------------+
         v                             v
   VISUALISE                     QUANTIFY  (needs a standards acquisition)
   imageR                        summarise_cal_levels
   quantile_hm                          v
                                 create_cal_curve
                                        v
                                 int2conc -> "pg_pixel", "pg_mm2"
                                        v
                                 plot_cal_coverage   (does calibration cover the data?)

  WHOLE STUDY:  run_study() (YAML)  .  generate_txt_images()  .  run_example()

Per-feature images can also be exported as plain .txt intensity matrices, which load directly into ImageJ and similar tools. Useful for downstream multimodal analysis such as co-registration with histology or other imaging modalities.

Quantification against calibration standards is covered separately in the Quantification with calibration standards vignette (vignette("quantification", package = "quantMSImageR")).

The workflow implemented here is described and applied in Smith et al., Analytical Chemistry (2024).

Installation

if (!requireNamespace("BiocManager", quietly = TRUE))
    install.packages("BiocManager")
BiocManager::install("quantMSImageR")

Quick start

The fastest way to see the whole workflow is run_example(), which runs the imaging workflow on two synthetic groups (A, circular tissue, n = 3; B, square tissue, n = 3), renders the HTML report, and also demonstrates quantification on the bundled calibration standards:

library(quantMSImageR)
res <- run_example()          # both groups; imaging report + calibration demo
res$calibrated                # the calibrated object

The rest of this vignette reproduces the key steps by hand.

The data model

Every acquisition is a Cardinal::MSImagingExperiment. quantMSImageR adds the quant_MSImagingExperiment subclass, which carries two extra slots for calibration and tissue metadata, so most functions coerce with as() first.

The object is features x pixels: one row per MRM transition, one column per pixel. Calculated quantities – response, snr, calibrated amount – are stored as additional spectra layers. Masking functions are the exception: applySNR(), back2NA() and zero2na() modify the selected val_slot in place, so keep a copy of the object when the unmodified values are needed afterwards.

Component Contains Accessor
spectra slots intensity (raw), response (IS-normalised), snr, pg_pixel / pg_mm2 (calibrated) spectra(x, "name"), spectraData(x)
feature metadata mz, name, transition m/z, analyte type, ion-library columns fData(x)
pixel metadata x, y, run, tissue/background label pData(x)
calibration fitted models, calibration levels, fit diagnostics calibrationModels(x), calibrationLevels(x), calibrationDiagnostics(x)
tissue summaries pixel and ROI matrices tissueMatrix(x), tissueData(x)

Multiple acquisitions are held in one object, distinguished by the run column of pData() – that is what combine_MSIs() produces and what every per-sample summary groups on.

Pixel metadata uses consistent vocabulary: run identifies the acquisition, x/y are grid coordinates, and the tissue mask lives in sample_name as tissue_pixels / background_pixels. The calibration path uses sample_type with Cal / Tissue / Background.

library(quantMSImageR)

p   <- system.file("extdata", "example.raw", "section01.RDS",
                   package = "quantMSImageR")
obj <- as(readRDS(p), "quant_MSImagingExperiment")

dim(obj)                        # features x pixels
#> [1]   8 400
names(spectraData(obj))         # available layers
#> [1] "intensity"
DT::datatable(as.data.frame(fData(obj)), rownames = FALSE,
              options = list(pageLength = 5, scrollX = TRUE))

Table 1. Feature metadata: one row per MRM transition.

DT::datatable(head(as.data.frame(pData(obj)), 100), rownames = FALSE,
              options = list(pageLength = 5, scrollX = TRUE))

Table 2. Pixel metadata, first 100 pixels of one section.

Reading your own data

The bundled example ships as .RDS so the vignette is reproducible, but real acquisitions enter through read_mrm(), which reads a Waters DESI-MRM .raw folder and joins the ion library to the features:

obj <- read_mrm(
  name         = "my_acquisition",        # .raw folder name, without the suffix
  folder       = "path/to/raw",           # directory containing my_acquisition.raw
  lib_ion_path = "path/to/ion_library.csv"
)

The .raw folder must contain the exported per-transition text files (an imaging/ subdirectory) or a previously cached MSImagingExperiment.rds. The ion library is a CSV with one row per transition; the columns used for matching are the precursor and product m/z, plus transition_id for the display name and Type to mark internal standards. Any additional columns – pathway class, polarity, collision energy – are carried through to fData() and become available for grouping, as shown in Joining ion-library metadata below.

The result is a features x pixels object exactly like the one inspected above, ready for select_tissue_pixels() and the rest of the workflow.

Loading the acquisitions

Families: acquisition, combining acquisitions.

Function Takes Produces
read_mrm() .raw folder + ion-library CSV MSImagingExperiment, fData joined to the library
trim_MSI() object with a tissue mask object with pure-background border rows/columns dropped
remove_blank_mzs() object object with features that have no data removed
align_features() two objects both reduced to common features, m/z-keyed
combine_MSIs() two or more aligned objects one object, each acquisition its own run
bind_panels() two objects on a shared (x, y) grid features of both on the pixel-grid intersection

Note the difference between the last two. combine_MSIs() stacks different sections side by side, adding pixels. bind_panels() merges two acquisitions of the same physical area – typically a positive- and a negative-mode run of one section, or two MRM panels – adding features to the same pixels:

pos <- read_mrm("acq_section1_pos", folder = "path/to/raw",
                lib_ion_path = "path/to/ion_library.csv")
neg <- read_mrm("acq_section1_neg", folder = "path/to/raw",
                lib_ion_path = "path/to/ion_library.csv")

# Pixels are matched on (x, y), so the two runs need not have identical grids;
# pixels present in only one are dropped.
both <- bind_panels(pos, neg, label = "S1")

The bundled example is six sections: three of group A (circular tissue) and three of group B (square tissue). They ship as RDS, so read_mrm() is not needed here. combine_MSIs() binds them into a single object, keeping each section as its own run:

library(quantMSImageR)
library(ggplot2)

sections <- sprintf("section%02d", 1:6)
objs <- lapply(sections, function(s)
  readRDS(system.file("extdata", "example.raw", paste0(s, ".RDS"),
                      package = "quantMSImageR")))
combined <- do.call(combine_MSIs, objs)

groups <- c("A", "A", "A", "B", "B", "B")   # sections 01-03 = A, 04-06 = B
combined
#> quant_MSImagingExperiment with 8 features and 2400 spectra 
#> spectraData(1): intensity
#> featureData(6): mz, feature_type, precursor_mz, product_mz, name, IS_norm
#> pixelData(4): x, y, run, sample_name
#> coord(2): x = 1...20, y = 1...20
#> runNames(6): section01, section02, section03, section04, section05, section06
#> experimentData(1): pixelSize
#> mass range: 1 to 8 
#> centroided: NA

Each section contributed its pixels as a separate run, on the same feature axis:

dim(combined)                       # features x (pixels from all six sections)
#> [1]    8 2400
table(pData(combined)$run)
#> 
#> section01 section02 section03 section04 section05 section06 
#>       400       400       400       400       400       400

Everything below operates on this one combined object.

Selecting tissue pixels

Family: acquisition.

Function Takes Produces
select_tissue_pixels() acquisition name, data path, ion library tissue_pixels.csv (x, y, tissue_pixels, background_pixels) plus a preview of the saved mask

Before SNR filtering, each acquisition needs a tissue mask. select_tissue_pixels() displays every feature’s ion image so you can pick the one that best separates tissue from background, then opens an interactive ROI selector:

select_tissue_pixels(
  name         = "my_acquisition",          # .raw folder name (no suffix)
  data_path    = "path/to/raw",
  lib_ion_path = "path/to/ion_library.csv"
)

It writes the resulting mask as tissue_pixels.csv inside the acquisition’s .raw folder, where int2snr() and generate_txt_images() read it. The mask is simply a per-pixel label – tissue versus background – which the bundled sections already carry in their sample_name column:

mask_tbl <- as.data.frame(pData(combined))[, c("run", "x", "y", "sample_name")]

DT::datatable(head(mask_tbl, 500), rownames = FALSE,
              options = list(pageLength = 5, scrollX = TRUE))

Table 3. Per-pixel tissue/background labelling used by int2snr() (first 500 pixels).

Plotting that column over the pixel coordinates shows the mask itself, which is the quickest way to confirm a selection before running the workflow – the circular group A and square group B tissue should stand out cleanly from the surrounding background:

ggplot(mask_tbl, aes(x, -y, fill = sample_name)) +
  geom_tile() +
  facet_wrap(~ run, ncol = 3) +
  coord_equal() +
  scale_fill_manual(values = c(tissue_pixels     = "#1b7837",
                               background_pixels = "grey85")) +
  labs(x = NULL, y = NULL, fill = NULL) +
  theme_minimal() +
  theme(axis.text = element_blank(), panel.grid = element_blank(),
        strip.text = element_text(size = 7),
        legend.text = element_text(size = 7))
Tissue masks for all six sections. Green pixels are labelled tissue_pixels and contribute to every downstream summary; grey pixels are background_pixels and are used to estimate the noise level in int2snr().

Tissue masks for all six sections. Green pixels are labelled tissue_pixels and contribute to every downstream summary; grey pixels are background_pixels and are used to estimate the noise level in int2snr().

Ion images

Family: visualisation.

Function Takes Produces
imageR() object, feat_ind, val_slot, sample_lab, scale a ggplot ion image
quantile_hm() object, quant_val, sample order/labels, feature_split a ComplexHeatmap::Heatmap (rows = samples, columns = features)
quant_palettes() palette name, optional n the hex colours used by both, for reuse in your own plots

Colours are consistent across the package and selectable per plot. imageR() takes palette = "heatmap0" (the default) or "viridis"; quantile_hm() takes a diverging palette plus group_palette / feature_palette for the metadata bars, and cell_border for the hairline that keeps neighbouring cells of the same colour from merging into one block. In a YAML-driven study these are set once in the colours: block:

colours:
  ion_image:   "heatmap0"  # heatmap0 | viridis
  heatmap:     "heatmap2"  # heatmap2 | heatmap0
  group:       "hat"       # hat | reading | heatmap0
  feature:     "reading"   # reading | hat | heatmap0
  cell_border: "white"     # any colour, or "none"
names(quant_palettes())
#> [1] "heatmap0" "heatmap2" "hat"      "reading"
quant_palettes("heatmap0")
#> [1] "#001219" "#005F73" "#0A9396" "#94D2BD" "#E9D8A6" "#EE9B00" "#CA6702"
#> [8] "#AE2012" "#9B2226"

imageR() returns a ggplot ion image for a chosen feature. Because it is a plain ggplot, the layout can be adjusted – here into a 3 x 2 grid:

imageR(combined, feat_ind = 1, sample_lab = "run", scale = "suppress",
       palette = "heatmap0") +
  facet_wrap(~ sample, ncol = 3) +
  theme(strip.text = element_text(size = 7),
        legend.text = element_text(size = 7))
Ion image of the first feature across all six sections, before SNR filtering. Sections 01-03 are group A (circular tissue), 04-06 are group B (square).

Ion image of the first feature across all six sections, before SNR filtering. Sections 01-03 are group A (circular tissue), 04-06 are group B (square).

Colour-scale treatment

A single ion image can look very different depending on how the colour scale is built, and the right choice depends on what is being judged. scale offers three treatments:

  • "suppress" caps the scale at percentile (default 99), so a few hot pixels cannot flatten everything else. This is the default and the right starting point for comparing sections.
  • "histogram" equalises the distribution, which reveals structure in a feature whose signal spans a narrow range – at the cost of a colour scale that no longer maps linearly to response.
  • "sqrt" compresses the top of the range more gently than histogram equalisation, keeping the scale monotonic in response.
.one <- combined[, pData(combined)$run == "section01"]
.p <- lapply(c("suppress", "histogram", "sqrt"), function(sc)
  imageR(.one, feat_ind = 1, sample_lab = "run", scale = sc) +
    labs(title = sc) +
    theme(legend.position = "none",
          plot.title = element_text(size = 9, face = "bold"),
          strip.text = element_blank()))
patchwork::wrap_plots(.p, nrow = 1)
The same feature and section under the three colour-scale treatments. Only `suppress` keeps the colour scale linear in response; the others trade that for visible structure.

The same feature and section under the three colour-scale treatments. Only suppress keeps the colour scale linear in response; the others trade that for visible structure.

perc_scale = TRUE additionally relabels the scale as a percentage of each image’s maximum, which is useful when the absolute numbers are not comparable anyway.

The HTML report produced by run_study() shows a related set of three views per feature, built with Cardinal’s own image() rather than imageR(): capped at the 95th quantile, histogram-normalised, and unscaled.

The same per-feature images can be written to disk as .txt intensity matrices (generate_txt_images(output_txt = TRUE), or output_txt: true in the YAML config). Those files open directly in ImageJ, which is the usual route into multimodal workflows.

Joining ion-library metadata

build_feature_meta() joins ion-library annotation to features by rounded (precursor, product) m/z, so extra columns – such as the pathway class Met-1 – become available for grouping. We do this before the heatmap because the heatmap uses Met-1 to split its rows.

The ion library is the CSV you supply as lib_ion_path, and it is worth looking at before anything else: it defines the panel. transition_id, precursor_mz, product_mz, collision_eV, cone_V, Polarity and Type are required; anything else is yours to add. Here Met-1 carries the pathway class and IS_norm names the internal standard each analyte is normalised to (see [int2response()]).

lib <- read.csv(system.file("extdata", "example_ion_library.csv",
                            package = "quantMSImageR"), check.names = FALSE)

DT::datatable(lib, rownames = FALSE,
              options = list(pageLength = 10, scrollX = TRUE))

Table 4. The example ion library, as supplied on disk.

fm <- build_feature_meta(combined, lib)

DT::datatable(fm[, c("name", "precursor_mz", "product_mz", "Type", "Met-1",
                     "IS_norm")],
              rownames = FALSE,
              options = list(pageLength = 10, scrollX = TRUE))

Table 5. The same metadata after build_feature_meta() has joined it to each measured feature by m/z.

Quantile heatmaps across samples

quantile_hm() summarises each feature to a per-sample quantile and returns a ComplexHeatmap::Heatmap. Rows are samples, grouped by study group; columns are features, split by the Met-1 class joined above – studies usually have more samples than features, so the long dimension runs vertically. Cells are drawn square (stretching to at most 1.5:1 when one dimension is much longer), so the heatmap grows with the data rather than distorting to fill the device.

cell_size sets that size in millimetres and fontsize the label size, which is what to reach for when long transition names crowd the panel: enlarge the cells and shrink the text rather than letting the labels dictate the layout.

The quant_val argument chooses which quantile summarises each feature – a median (0.5) reflects the bulk of the tissue, while a high quantile (0.95) emphasises hot-spots:

feature_split <- fm[["Met-1"]]

quantile_hm(combined, quant_val = 0.5,
            heatmap_order = sections, heatmap_labs = groups,
            feature_split = feature_split, feature_split_name = "Met-1",
            cell_size = 10, fontsize = 7)
Median (50th percentile) feature intensity per section. Rows are sections grouped by study group; columns are features split by Met-1 class.

Median (50th percentile) feature intensity per section. Rows are sections grouped by study group; columns are features split by Met-1 class.

quantile_hm(combined, quant_val = 0.95,
            heatmap_order = sections, heatmap_labs = groups,
            feature_split = feature_split, feature_split_name = "Met-1",
            cell_size = 10, fontsize = 7)
The same summary at the 95th percentile, which emphasises hot-spots rather than the bulk of the tissue.

The same summary at the 95th percentile, which emphasises hot-spots rather than the bulk of the tissue.

Signal-to-noise filtering

Family: filtering.

Function Takes Produces
int2response() object + internal-standard name intensities normalised to the IS (response)
int2snr() object, background/tissue labels, snr_thresh, optional snr_overrides adds an snr slot; below threshold and non-tissue become NA
applySNR() object carrying an snr slot sub-threshold pixels set to NA in val_slot
back2NA() object + background label background pixels set to NA
zero2na() object, val_slot zeros replaced with NA

int2snr() computes a per-pixel signal-to-noise ratio for each feature against the background pixels, and applySNR() then sets every sub-threshold pixel to NA in the intensity slot.

combined <- int2snr(combined, val_slot = "intensity", snr_thresh = 3)
combined <- applySNR(combined, val_slot = "intensity")

int2snr() added an snr layer rather than replacing anything, and applySNR() then set the sub-threshold pixels of intensity to NA:

names(spectraData(combined))                            # snr layer added
#> [1] "intensity" "snr"
mean(is.na(spectra(combined, "intensity"))) |> round(3) # fraction now masked
#> [1] 0.625

Compare the filtered image below with the unfiltered ion images in Ion images above: the low-signal background has been removed, leaving the tissue.

imageR(combined, feat_ind = 1, sample_lab = "run", val_slot = "intensity",
       scale = "suppress") +
  facet_wrap(~ sample, ncol = 3) +
  theme(strip.text = element_text(size = 7),
        legend.text = element_text(size = 7))
The same feature after applySNR(). Sub-threshold pixels are now NA, so the low-signal background has been removed and only tissue signal remains (compare to previous ion images).

The same feature after applySNR(). Sub-threshold pixels are now NA, so the low-signal background has been removed and only tissue signal remains (compare to previous ion images).

Running a full study from a YAML config

Family: workflow.

Function Takes Produces
run_study() a YAML configuration file runs the whole study: .txt images, optional calibration, one HTML report per SNR threshold
validate_config() the same file a report of every problem found in it, before anything is read or written
generate_txt_images() acquisition names, paths, ion library, SNR settings list of combined/SNR-filtered objects; optional per-feature .txt images
run_example() nothing (all paths resolved internally) the same list plus calibrated and report, having rendered the HTML report

Everything above runs inside generate_txt_images(), which run_study() drives from a YAML file. In practice a whole study – multiple acquisitions, SNR sweeps, ion ratios and optional calibration – is configured in one place. A minimal config looks like:

study: "my_study"
paths:
  data_path:    "path/to/raw"
  out_path:     "path/to/results"
  image_dir:    "path/to/images"
  lib_ion_path: "path/to/ion_library.csv"
samples:
  - pos: "acq_ctrl"
    run_id: "S1"
    label:  "Ctrl"
  - pos: "acq_treated"
    run_id: "S2"
    label:  "Treatment"
parameters:
  snr_thresh: [3]
output:
  render_report: true
  output_txt:    true      # write per-feature .txt images for ImageJ

The shipped template documents every option (SNR overrides, ratios, calibration, …) – open it to use as a starting point:

file.show(system.file("config_template.yaml", package = "quantMSImageR"))

Then point run_study() at your filled-in config:

run_study("path/to/study_config.yaml")

The same runner is available from the command line, which is the usual way to run a large study unattended:

Rscript -e 'quantMSImageR::run_study("study_config.yaml")'

The configuration as a record of the analysis

Driving a study from a file rather than from a session means a complete human-readable configuration file to document how the data were processed. Every choice that affects the results (which acquisitions belong to which group, the SNR thresholds and any per-analyte overrides, whether internal-standard normalisation ran and against which standard, the calibration design and its weighting, how much extrapolation was tolerated) is stored for future reference.

The file is small enough to deposit alongside the data, cite in a methods section, or track in version control in line with FAIR principles (Wilkinson et al., 2016).

To ensure accuracy of this documentation validate_config() runs every check rather than stopping at the first failure, so one call reports every problem in the file:

validate_config("path/to/study_config.yaml")
quantMSImageR validation: 2 error(s), 1 warning(s), 26 check(s)

Errors:
  - parameters$average_method = 'geometric'; use 'mean' or 'median'.
  - parameters$is_name = 'PGE2-d4' is not a value of the ion library's 'Type'
    column. Values present: Analyte, IS. Note this is a type label, not a
    transition name.

run_study() calls it first and stops if there are errors, so a configuration that cannot be trusted never produces an output. The checks are deliberately cross-file for every enabled functionality (the ion library, the calibration metadata and the config have to agree with each other, not merely be individually well-formed).

References

Smith MJ, Nie M, Adner M, Säfholm J, Wheelock CE. Development of a Desorption Electrospray Ionization–Multiple-Reaction-Monitoring Mass Spectrometry (DESI-MRM) Workflow for Spatially Mapping Oxylipins in Pulmonary Tissue. Analytical Chemistry (2024). https://doi.org/10.1021/acs.analchem.4c02350

Wilkinson MD, et al. The FAIR Guiding Principles for scientific data management and stewardship. Scientific Data 3, 160018 (2016). https://doi.org/10.1038/sdata.2016.18

Session information

sessionInfo()
#> R version 4.6.1 (2026-06-24)
#> Platform: x86_64-pc-linux-gnu
#> Running under: Ubuntu 26.04 LTS
#> 
#> Matrix products: default
#> BLAS:   /usr/lib/x86_64-linux-gnu/openblas-pthread/libblas.so.3 
#> LAPACK: /usr/lib/x86_64-linux-gnu/openblas-pthread/libopenblasp-r0.3.32.so;  LAPACK version 3.12.0
#> 
#> locale:
#>  [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
#>  [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
#>  [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
#>  [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
#>  [9] LC_ADDRESS=C               LC_TELEPHONE=C            
#> [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       
#> 
#> time zone: Etc/UTC
#> tzcode source: system (glibc)
#> 
#> attached base packages:
#> [1] stats4    stats     graphics  grDevices utils     datasets  methods  
#> [8] base     
#> 
#> other attached packages:
#> [1] quantMSImageR_0.99.0 Cardinal_3.15.0      S4Vectors_0.51.5    
#> [4] ProtGenerics_1.45.0  BiocGenerics_0.59.10 generics_0.1.4      
#> [7] BiocParallel_1.47.0  ggplot2_4.0.3        BiocStyle_2.41.0    
#> 
#> loaded via a namespace (and not attached):
#>  [1] gtable_0.3.6          circlize_0.4.18       shape_1.4.6.1        
#>  [4] rjson_0.2.23          xfun_0.60             bslib_0.11.0         
#>  [7] htmlwidgets_1.6.4     GlobalOptions_0.1.4   Biobase_2.73.1       
#> [10] lattice_0.22-9        vctrs_0.7.3           tools_4.6.1          
#> [13] crosstalk_1.2.2       parallel_4.6.1        tibble_3.3.1         
#> [16] cluster_2.1.8.2       pkgconfig_2.0.3       Matrix_1.7-5         
#> [19] RColorBrewer_1.1-3    S7_0.2.2              lifecycle_1.0.5      
#> [22] compiler_4.6.1        farver_2.1.2          codetools_0.2-20     
#> [25] ComplexHeatmap_2.29.0 clue_0.3-68           htmltools_0.5.9      
#> [28] sys_3.4.3             buildtools_1.0.0      sass_0.4.10          
#> [31] yaml_2.3.12           crayon_1.5.3          pillar_1.11.1        
#> [34] jquerylib_0.1.4       tidyr_1.3.2           DT_0.34.0            
#> [37] cachem_1.1.0          magick_2.9.1          iterators_1.0.14     
#> [40] viridis_0.6.5         foreach_1.5.2         nlme_3.1-170         
#> [43] tidyselect_1.2.1      digest_0.6.39         dplyr_1.2.1          
#> [46] purrr_1.2.2           labeling_0.4.3        maketools_1.3.2      
#> [49] fastmap_1.2.0         grid_4.6.1            colorspace_2.1-3     
#> [52] cli_3.6.6             magrittr_2.0.5        patchwork_1.3.2      
#> [55] utf8_1.2.6            withr_3.0.3           matter_2.15.0        
#> [58] scales_1.4.0          chemCal_0.2.3         rmarkdown_2.31       
#> [61] matrixStats_1.5.0     otel_0.2.0            gridExtra_2.3.1      
#> [64] GetoptLong_1.1.1      png_0.1-9             evaluate_1.0.5       
#> [67] knitr_1.51            IRanges_2.47.2        doParallel_1.0.17    
#> [70] irlba_2.3.7           viridisLite_0.4.3     CardinalIO_1.11.0    
#> [73] rlang_1.3.0           Rcpp_1.1.2            ontologyIndex_2.12   
#> [76] glue_1.8.1            BiocManager_1.30.27   jsonlite_2.0.0       
#> [79] R6_2.6.1