From 05adeb7c8bee8c688165c441fe6074b326d9fe51 Mon Sep 17 00:00:00 2001 From: "chris.bielow@fu-berlin.de" Date: Fri, 11 Apr 2025 16:02:39 +0200 Subject: [PATCH 1/5] + vastly rewrite map alignment tutorial + add more glossary terms + extend feature linking tutorial + add new PSM_to_features tutorial for IDMapper --- docs/source/user_guide/PSM_to_features.rst | 88 + docs/source/user_guide/feature_linking.rst | 111 +- docs/source/user_guide/glossary.rst | 12 +- docs/source/user_guide/img/map_alignment.png | Bin 73858 -> 0 bytes .../user_guide/img/map_alignment_fmaps.png | Bin 0 -> 120013 bytes .../user_guide/img/map_alignment_trafos.png | Bin 0 -> 67897 bytes docs/source/user_guide/index.rst | 1 + docs/source/user_guide/map_alignment.rst | 261 +- docs/source/user_guide/quantitative_data.rst | 4 +- src/data/BSA1_F1.idXML | 172 + src/data/BSA1_F1_idmapped.featureXML | 16952 ++++++++-------- src/data/BSA2_F1_idmapped.featureXML | 8296 ++++++++ src/data/BSA3_F1_idmapped.featureXML | 6898 +++++++ 13 files changed, 24244 insertions(+), 8551 deletions(-) create mode 100644 docs/source/user_guide/PSM_to_features.rst delete mode 100644 docs/source/user_guide/img/map_alignment.png create mode 100644 docs/source/user_guide/img/map_alignment_fmaps.png create mode 100644 docs/source/user_guide/img/map_alignment_trafos.png create mode 100644 src/data/BSA1_F1.idXML create mode 100644 src/data/BSA2_F1_idmapped.featureXML create mode 100644 src/data/BSA3_F1_idmapped.featureXML diff --git a/docs/source/user_guide/PSM_to_features.rst b/docs/source/user_guide/PSM_to_features.rst new file mode 100644 index 000000000..0581a236c --- /dev/null +++ b/docs/source/user_guide/PSM_to_features.rst @@ -0,0 +1,88 @@ +Feature Map Annotation with Peptide Identifications +=================================================== + +A :term:`feature map` is usually obtained from running a :term:`feature finder`, e.g. :py:class:`~.FeatureFinderAlgorithmPicked` (see `Feature Detection `_). +A logical next step is to compare features across runs using `Map Alignment `_ and `Feature Linking `_. +However, most map aligners in pyOpenMS require features which are annotated with PSMs (see `Identication Data `_). +To link features to their respective PSMs (as obtained from a search engine, such as Comet), we can use the :py:class:`~.IDMapper`. + + +Step 0: Download Example data +----------------------------- + +.. code-block:: python + :linenos: + + import pyopenms as oms + from urllib.request import urlretrieve + + base_url = ( + "https://raw.githubusercontent.com/OpenMS/pyopenms-docs/master/src/data/" + ) + + feature_file = "BSA1_F1.featureXML" + urlretrieve(base_url + feature_file, feature_file) + + idxml_file = "BSA1_F1.idXML" + urlretrieve(base_url + idxml_file, idxml_file) + +Step 1: Load the Feature Map +---------------------------- + +First, load the FeatureMap from a `.featureXML` file: + +.. code-block:: python + + import pyopenms as oms + + feature_map = oms.FeatureMap() + oms.FeatureXMLFile().load(feature_file, feature_map) + +Step 2: Load Peptide Identifications +------------------------------------ + +Next, load the PeptideIdentifications from an `.idXML` file: + +.. code-block:: python + + peptide_ids = [] + protein_ids = [] + oms.IdXMLFile().load(idxml_file, protein_ids, peptide_ids) + +Step 3: Initialize and Configure `IDMapper` +------------------------------------------- + +Now, configure `IDMapper` to apply **retention time (RT) and m/z tolerance settings**: + +.. code-block:: python + + id_mapper = oms.IDMapper() + params = id_mapper.getParameters() + params.setValue("rt_tolerance", 5.0) # RT tolerance in seconds + params.setValue("mz_tolerance", 10.0) # m/z tolerance in ppm + id_mapper.setParameters(params) + +Step 4: Annotate the FeatureMap +------------------------------- + +Use the configured `IDMapper` to link peptide IDs to the FeatureMap: + +.. code-block:: python + + id_mapper.annotate(feature_map, peptide_ids, protein_ids, use_centroid_rt=True, use_centroid_mz=True, spectra=None) + +Step 5: Save the Annotated FeatureMap +-------------------------------------- + +Finally, store the modified FeatureMap back to a file: + +.. code-block:: python + + oms.FeatureXMLFile().store("BSA1_F1_annotated.featureXML", feature_map) + +.. tip:: + You can visualize the annotated FeatureMap using OpenMS visualization tools like `TOPPView`. + + +You have successfully **annotated a FeatureMap** with PeptideIdentifications using `IDMapper`. This allows further downstream analysis in (py)OpenMS workflows. + diff --git a/docs/source/user_guide/feature_linking.rst b/docs/source/user_guide/feature_linking.rst index f408816c5..34cdbcca8 100644 --- a/docs/source/user_guide/feature_linking.rst +++ b/docs/source/user_guide/feature_linking.rst @@ -14,17 +14,17 @@ are available in pyOpenMS: - :py:class:`~.FeatureGroupingAlgorithmQT` - :py:class:`~.FeatureGroupingAlgorithmKD` -- :py:class:`~.FeatureGroupingAlgorithm` - :py:class:`~.FeatureGroupingAlgorithmLabeled` - :py:class:`~.FeatureGroupingAlgorithmUnlabeled` -We now perform a features linking using the :py:class:`~.FeatureGroupingAlgorithmQT` algorithm. +We now perform feature linking using the :py:class:`~.FeatureGroupingAlgorithmQT` algorithm. Download Example Data ********************* .. code-block:: python - + :linenos: + import pyopenms as oms from urllib.request import urlretrieve @@ -47,28 +47,115 @@ Download Example Data oms.FeatureXMLFile().load(feature_file, feature_map) feature_maps.append(feature_map) -features Linking Algorithm +Feature Linking Algorithm ****************************************** -All :py:class:`~.FeatureMap` objects will be combined in a :py:class:`~.ConsensusMap`. +Feature linking is the process which connects (links) features with similar RT and m/z across different runs. +This enables comparisons of peptide signals across runs and is a prerequisite for downstream statistical analysis. +Thus, all features across all :py:class:`~.FeatureMap` objects will be combined into a :py:class:`~.ConsensusMap`. +Here we use :py:class:`~.FeatureGroupingAlgorithmQT` to find corresponding features with similar RT and m/z across feature maps. +If everything goes well, we will have a lot of triplets in our test run (we have have three input maps; one feature from each feature map) within each consensus feature. +Realistically, some features will remain unmatched, forming singletons or doublets—consensus features of size 1 or 2. These undersized consensus features can arise due to several factors. In some cases, the signal may be genuinely absent in the dataset, preventing feature detection. Alternatively, the feature-finding algorithm might fail to identify the feature in all available maps, leading to incomplete linkage. Additionally, suboptimal parameters for feature linking—such as excessive retention time (RT) variations across samples—can contribute to mismatches, further reducing the likelihood of forming larger consensus features. The latter can be corrected for using a process called map alignment, which we adressed in a `previous chapter `_. + .. code-block:: python + :linenos: + + consensus_map = oms.ConsensusMap() + # populate the actual cmap using FeatureGroupingAlgorithmQT feature_grouper = oms.FeatureGroupingAlgorithmQT() + # this is where the work happens: + feature_grouper.group(feature_maps, consensus_map) - consensus_map = oms.ConsensusMap() +That's it! +Now, we can print some summary information: + + +.. code-block:: python + :linenos: + + # print some info on the consensus map + print(f"Total number of consensus features: {consensus_map.size()}\n\n") + + from collections import defaultdict + + def compute_feature_size_stats(consensus_map): + size_counts = defaultdict(int) # Default value for missing keys is 0 + + for cfeature in consensus_map: + size = len(cfeature.getFeatureList()) + size_counts[size] += 1 # No need to check if the key exists + + return size_counts + + stats = compute_feature_size_stats(consensus_map) + ## how many consensus features are there which contain features from 1, 2, or all 3 feature maps? + print("ConsensusFeature Size Distribution:", dict(stats), "\n\n") + + for (i, cfeature) in enumerate(consensus_map): + if i > 3: + break + print( + f"ConsensusFeature: average int: {cfeature.getIntensity()}, RT: {cfeature.getRT()}, m/z: {cfeature.getMZ()}" + ) + # The two features in map 1 and map 2 represent the same analyte at + # slightly different RT and m/z + for fh in cfeature.getFeatureList(): + print(f" -- Feature: map#: {fh.getMapIndex()}, int: {fh.getIntensity()}, RT: {fh.getRT()}") + + +This then prints: + +.. code-block:: output + + Map 0: Filename = BSA1_F1.featureXML, Size = 256, UniqueID = 15391035140702131759 + Map 1: Filename = BSA2_F1.featureXML, Size = 235, UniqueID = 4409371154349730995 + Map 2: Filename = BSA3_F1.featureXML, Size = 204, UniqueID = 16851873442542870272 + Total number of consensus features: 474 + + + ConsensusFeature Size Distribution: {3: 55, 2: 111, 1: 308} + + + ConsensusFeature: average int: 674365.6875, RT: 1925.75138611056, m/z: 387.7128198087733 + -- Feature: map#: 0, int: 1432460.0, RT: 1961.42395727216 + -- Feature: map#: 1, int: 285228.0, RT: 1914.26950841529 + -- Feature: map#: 2, int: 305409.0, RT: 1901.56069264423 + ConsensusFeature: average int: 1154167.0, RT: 1787.5601793634498, m/z: 662.2911690419279 + -- Feature: map#: 0, int: 834075.0, RT: 1836.92005770327 + -- Feature: map#: 1, int: 2119760.0, RT: 1748.02632036743 + -- Feature: map#: 2, int: 508666.0, RT: 1777.73416001965 + ConsensusFeature: average int: 562620.0, RT: 1650.6520418306134, m/z: 779.7355985805051 + -- Feature: map#: 0, int: 661665.0, RT: 1662.1136104186 + -- Feature: map#: 1, int: 751397.0, RT: 1616.78250841374 + -- Feature: map#: 2, int: 274798.0, RT: 1673.0600066595 + ConsensusFeature: average int: 23445466.0, RT: 1877.1572820450535, m/z: 738.310987605286 + -- Feature: map#: 0, int: 34988800.0, RT: 1901.57762589719 + -- Feature: map#: 1, int: 25276700.0, RT: 1849.00759248543 + -- Feature: map#: 2, int: 10070900.0, RT: 1880.88662775254 + +The results here are not ideal, since we only found 55 features which are present in all three feature maps. The remaining consensus features have a missing value for at least one feature map. +We could tweak the parameters used for grouping, but you should make sure that the, for example, acceptable RT deltas are sensible. There is a tradeoff between sensitivity and specificity here. + +Finally, we add some meta-data to the consensus map, which allows us to track the input data later on if we were to store the consensus map to disk. + +.. code-block:: python + :linenos: file_descriptions = consensus_map.getColumnHeaders() - # collect information about input maps + # collect information about input maps ... for i, feature_map in enumerate(feature_maps): file_description = file_descriptions.get(i, oms.ColumnHeader()) - file_description.filename = feature_map.getDataProcessing()[0].getMetaValue( - "parameter: in" - )[:-5] + file_description.filename = feature_files[i] ## filename file_description.size = feature_map.size() file_description.unique_id = feature_map.getUniqueId() file_descriptions[i] = file_description - + # ... and store them in the column headers of the cmap consensus_map.setColumnHeaders(file_descriptions) - feature_grouper.group(feature_maps, consensus_map) + + # print the information on underlying feature maps which we just collected + file_descriptions = consensus_map.getColumnHeaders() + for index, header in file_descriptions.items(): + print(f"Map {index}: Filename = {header.filename}, Size = {header.size}, UniqueID = {header.unique_id}") diff --git a/docs/source/user_guide/glossary.rst b/docs/source/user_guide/glossary.rst index a99715488..35064339d 100644 --- a/docs/source/user_guide/glossary.rst +++ b/docs/source/user_guide/glossary.rst @@ -28,6 +28,7 @@ A glossary of common terms used throughout OpenMS documentation. One consensus map usually contains many consensus features. OpenMS represents a consensus map using the class `ConsensusMap `_. + de novo de novo peptide sequencing A peptide's amino acid sequence is inferred directly from the precursor peptide mass and tandem mass spectrum (:term:`MS2` or :term:`MS3`) fragment ions, without comparison to a reference proteome. @@ -45,6 +46,10 @@ A glossary of common terms used throughout OpenMS documentation. features A feature, in the OpenMS terminology, subsumes all m/z signals originating from a single compound at a certain charge state. This includes the isotope pattern and usually spans multiple spectra in retention time (the elution profile). + + feature finder + FeatureFinder + A FeatureFinder in pyOpenMS creates a :term:`feature map` using the MS1 spectra from a :term:`peak map` (e.g. from an mzML file) as input. feature maps feature map @@ -154,9 +159,10 @@ A glossary of common terms used throughout OpenMS documentation. peptide-spectrum match PSM - A method used in proteomics to identify proteins from a complex mixture. Involves comparing the - mass spectra of peptide fragments generated from a protein sample with a database of predicted - spectra, in order to identify the protein that produced the observed peptides. + PSMs + A peptide-spectrum match associates a peptide sequence (possibly including modifications) to an MS/MS spectrum. + This usually involves comparing the mass spectra of peptide fragments generated from a digested protein sample with a database of predicted + spectra. Alternatively, this can be done using :term:`de novo` techniques (without a database, just using observed spectra). 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Different map alignment algorithms are available in pyOpenMS: @@ -13,16 +18,56 @@ Different map alignment algorithms are available in pyOpenMS: - :py:class:`~.MapAlignmentAlgorithmPoseClustering` - :py:class:`~.MapAlignmentAlgorithmIdentification` - :py:class:`~.MapAlignmentAlgorithmKD` + +If you want to apply a custom RT mapping, you can use: + - :py:class:`~.MapAlignmentTransformer` -To perform a simple linear alignment we can employ the algorithm :py:class:`~.MapAlignmentAlgorithmPoseClustering`. -In the example below it is used for the alignment of :term:`feature maps`. +To perform a simple linear alignment (an affine transformation to be exact, i.e. an offset and a slope) we can employ the algorithm :py:class:`~.MapAlignmentAlgorithmPoseClustering`. +This algorithm in OpenMS can also align peak maps, but we usually recommend aligning on the feature level, since +it's much faster (less features than peaks) and also more stable. + +Other map alignment algorithms in pyOpenMS require the input data to have :term:`PSMs` annotated. See `Annotating PSMs to features `_ on how to do that. +Also, some require (or support) a reference map (the one which all the other feature maps align to). More on this below. +All algorithms except :py:class:`~.MapAlignmentAlgorithmPoseClustering` can correct the RT using a variety of models/functions fitted to the landmarks used across feature maps. + +Here is a summary table: + +.. list-table:: Comparison of pyOpenMS' map alignment algorithms + :header-rows: 1 + + * - algorithm + - input type + - transformation in RT + - uses & requires PSMs + - reference map + * - :py:class:`~.MapAlignmentAlgorithmPoseClustering` + - feature maps or peak maps + - linear (affine transformation) + - no + - required + * - :py:class:`~.MapAlignmentAlgorithmIdentification` + - peak map, feature maps, IDs + - any [#footnote1]_ + - yes + - optional + * - :py:class:`~.MapAlignmentAlgorithmKD` + - feature maps + - any [#footnote1]_ + - yes + - not supported/needed + +.. [#footnote1] Valid models are "linear","b_spline","lowess" or "interpolated" + +You should pick the algorithm which does not overfit your data and maximizes the number of complete consensus features during a later `feature linking `_ stage. + Download Example Data ********************* .. code-block:: python - + :linenos: + import pyopenms as oms from urllib.request import urlretrieve @@ -30,10 +75,12 @@ Download Example Data "https://raw.githubusercontent.com/OpenMS/pyopenms-docs/master/src/data/" ) + # we use featureXML files which already contain PSMs (as obtained by oms.IDMapper()) + # ... so we can use all aligners pyOpenMS has to offer feature_files = [ - "BSA1_F1.featureXML", - "BSA2_F1.featureXML", - "BSA3_F1.featureXML", + "BSA1_F1_idmapped.featureXML", + "BSA2_F1_idmapped.featureXML", + "BSA3_F1_idmapped.featureXML", ] feature_maps = [] @@ -45,14 +92,20 @@ Download Example Data oms.FeatureXMLFile().load(feature_file, feature_map) feature_maps.append(feature_map) -Map Alignment Algorithm -*********************** +Selecting a Reference Map +************************* + +Some alignment algorithms in pyOpenMS require to set a reference, e.g. :py:class:`~.MapAlignmentAlgorithmPoseClustering`. +For others, it is optional, e.g., :py:class:`~.MapAlignmentAlgorithmIdentification`. +Finally, :py:class:`~.MapAlignmentAlgorithmKD` is reference-free by definition. -From the list of :term:`feature maps`, the one with the largest number of features is selected for reference. +For :py:class:`~.MapAlignmentAlgorithmPoseClustering` we could pick the :term:`feature map` with the largest number of features as a reference, +to provide the maximum number of reference points. The retention times of the other :term:`feature maps` are aligned to this. .. code-block:: python - + :linenos: + # set ref_index to feature map index with largest number of features ref_index = [ i[0] @@ -62,77 +115,167 @@ The retention times of the other :term:`feature maps` are aligned to this. ][-1] aligner = oms.MapAlignmentAlgorithmPoseClustering() - aligner.setReference(feature_maps[ref_index]) + + ## push all maps except the reference into the align method; don't align the reference to itself for max. efficiency + feature_maps_to_align = feature_maps[:ref_index] + feature_maps[ref_index+1:] + + +Map Alignment Algorithm +*********************** + +Now, let's run an algorithm. Since the interfaces of the aligners differ slightly, we will go though them one by one, starting with :py:class:`~.MapAlignmentAlgorithmPoseClustering`. + +.. code-block:: python + :linenos: + aligner = oms.MapAlignmentAlgorithmPoseClustering() + aligner.setReference(feature_maps[ref_index]) + + + ## change default params, if you want ... + p = aligner.getParameters() + # p.setValue(...) + aligner.setParameters(p) + # perform alignment and transformation of feature maps to the reference map (exclude reference map) - for feature_map in feature_maps[:ref_index] + feature_maps[ref_index + 1 :]: + for feature_map in feature_maps_to_align: trafo = oms.TransformationDescription() aligner.align(feature_map, trafo) transformer = oms.MapAlignmentTransformer() transformer.transformRetentionTimes( feature_map, trafo, True - ) # store original RT as meta value + ) # store original RT as meta value in `feature_map` + + +The interface of :py:class:`~.MapAlignmentAlgorithmIdentification` is a bit different, so have to adapt our code a bit. +In contrast to :py:class:`~.MapAlignmentAlgorithmPoseClustering`, which always fits a linear model, we can choose which model to fit to the landmarks found by the algorithm. Our choices are "linear","b_spline","lowess" and "interpolated". + +.. code-block:: python + :linenos: + + aligner = oms.MapAlignmentAlgorithmIdentification() + ## we could set a reference map; but we don't. Instead, we rely on the algorithm to use an internal average of all maps + ref_index = -1 # -1 means 'take the median' for this algorithm + + ## let's change some default parameters of MapAlignmentAlgorithmIdentification, just to see how it's done: + p = aligner.getParameters() + p.setValue("max_rt_shift", 0.2) # 20% of total RT span + p.setValue("use_feature_rt", "true") + aligner.setParameters(p) + + ## this list will be filled with transformations; you could use them to transform the original mzML, for example + trafos = list() + aligner.align(feature_maps, trafos, ref_index) + + # the transformations now contain the landmarks, but we still need to compute (fit) a model to them, + # ... before applying it to the feature maps: + # Possible models are: "linear","b_spline","lowess","interpolated" + for fm, trafo in zip(feature_maps, trafos): + trafo.fitModel("linear") + transformer = oms.MapAlignmentTransformer() + transformer.transformRetentionTimes( + fm, trafo, True + ) # stores original RT as meta value 'original_RT' for each feature in each `feature_map` + + +You can test different models, but since we only have very few data points in our toy example, the linear model works best. Visualization ************* -Plotting :term:`consensus maps` with features before and after alignment. +Plotting :term:`feature maps` before and after alignment. We should observe that the data points cluster closer together: .. code-block:: python + :linenos: import matplotlib.pyplot as plt import numpy as np - fmaps = ( - [feature_maps[ref_index]] - + feature_maps[:ref_index] - + feature_maps[ref_index + 1 :] - ) + def plot_consensus_maps(fmaps): + """ + Plots consensus maps before and after alignment. + + Parameters: + - fmaps: List of FeatureMaps. + """ + fig, axes = plt.subplots(1, 2, figsize=(10, 5)) + + titles = ["consensus map before alignment", "consensus map after alignment"] + x_labels = ["RT", "RT"] + y_label = "m/z" + + for i, ax in enumerate(axes): + ax.set_title(titles[i]) + ax.set_xlabel(x_labels[i]) + if i == 0: ax.set_ylabel(y_label) + + for fm in fmaps: + x_data = [f.getMetaValue("original_RT") if i == 0 else f.getRT() for f in fm] + y_data = [f.getMZ() for f in fm] + alpha_values = np.asarray([f.getIntensity() for f in fm]) / max([f.getIntensity() for f in fm]) + ax.scatter(x_data, y_data, alpha=alpha_values) + + fig.tight_layout() + fig.legend( + [fmap.getDataProcessing()[0].getMetaValue("parameter: out")[:-11] for fmap in fmaps], + loc="lower center", + ) + plt.show() - fig = plt.figure(figsize=(10, 5)) + # Example usage: + plot_consensus_maps(feature_maps) - ax = fig.add_subplot(1, 2, 1) - ax.set_title("consensus map before alignment") - ax.set_ylabel("m/z") - ax.set_xlabel("RT") +.. image:: img/map_alignment_fmaps.png - # use alpha value to display feature intensity - ax.scatter( - [f.getRT() for f in fmaps[0]], - [f.getMZ() for f in fmaps[0]], - alpha=np.asarray([f.getIntensity() for f in fmaps[0]]) - / max([f.getIntensity() for f in fmaps[0]]), - ) - for fm in fmaps[1:]: - ax.scatter( - [f.getMetaValue("original_RT") for f in fm], - [f.getMZ() for f in fm], - alpha=np.asarray([f.getIntensity() for f in fm]) - / max([f.getIntensity() for f in fm]), - ) +We can also inspect the underlying transformation and the landmarks it is based upon: + +.. code-block:: python + :linenos: + + def plot_transformed_rt_with_trafo(fmaps, trafos): + """ + Plots original RT vs. transformed RT for each feature map and ensures matching colors for transformation points. - ax = fig.add_subplot(1, 2, 2) - ax.set_title("consensus map after alignment") - ax.set_xlabel("RT") + Parameters: + - fmaps: List of FeatureMaps that have been aligned. + - trafos: List of Transformations applied to the FeatureMaps. + """ + fig, ax = plt.subplots(figsize=(8, 5)) - for fm in fmaps: - ax.scatter( - [f.getRT() for f in fm], - [f.getMZ() for f in fm], - alpha=np.asarray([f.getIntensity() for f in fm]) - / max([f.getIntensity() for f in fm]), - ) + ax.set_title("RT Transformation Before vs. After Alignment") + ax.set_xlabel("Original RT") + ax.set_ylabel("Transformed RT") - fig.tight_layout() - fig.legend( - [ - fmap.getDataProcessing()[0].getMetaValue("parameter: out")[:-11] - for fmap in fmaps - ], - loc="lower center", - ) - fig.show() + colors = plt.cm.viridis(np.linspace(0, 1, len(fmaps))) # Generate distinct colors for each map + + for i, (fm, trafo) in enumerate(zip(fmaps, trafos)): + label = fm.getDataProcessing()[0].getMetaValue("parameter: out")[:-11] + + # Extract original and transformed RTs from feature maps + original_rt = [f.getMetaValue("original_RT") for f in fm] + transformed_rt = [f.getRT() for f in fm] + + # Plot feature map RTs + ax.scatter(original_rt, transformed_rt, alpha=0.5, s=20, color=colors[i], label=f"{label} - Features") + + # Extract transformation points + trafo_points = trafo.getDataPoints() + original_trafo_rt = [point.first for point in trafo_points] + transformed_trafo_rt = [point.second for point in trafo_points] + + # Plot transformation points + ax.scatter(original_trafo_rt, transformed_trafo_rt, marker="D", s=100, edgecolors="black", color=colors[i], + label=f"{label} - Trafo") + + ax.legend(loc="upper left") + ax.grid(True) + plt.show() + + # Example usage: + plot_transformed_rt_with_trafo(feature_maps, trafos) -.. image:: img/map_alignment.png +While we do not know the underlying features using just this information, we can see that features in corresponding maps are now nicely aligned horizontally (the new RT). Also, it seems a linear model nicely fits all the data points in each feature map. Using a more flexible model like LOWESS will likely result in a worse alignment (try and see for yourself). + +.. image:: img/map_alignment_trafos.png \ No newline at end of file diff --git a/docs/source/user_guide/quantitative_data.rst b/docs/source/user_guide/quantitative_data.rst index f4c4998e0..2e3b0d689 100644 --- a/docs/source/user_guide/quantitative_data.rst +++ b/docs/source/user_guide/quantitative_data.rst @@ -74,7 +74,9 @@ quantitative data directly and it supports direct iteration in Python: Often :term:`LC-MS/MS` experiments are run to compare quantitative features across experiments. In OpenMS, linked features from individual experiments are -represented by a :py:class:`~.ConsensusFeature` +represented by a :py:class:`~.ConsensusFeature`. +We will explore how :term:`Consensus Maps` are created in a process called FeatureLinking in the `Feature Linking `_ chapter. +For now, we focus on how to build :term:`consensus feature`s and their container (consensus maps) manually. .. code-block:: python :linenos: diff --git a/src/data/BSA1_F1.idXML b/src/data/BSA1_F1.idXML new file mode 100644 index 000000000..8952fdf63 --- /dev/null +++ b/src/data/BSA1_F1.idXML @@ -0,0 +1,172 @@ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + diff --git a/src/data/BSA1_F1_idmapped.featureXML b/src/data/BSA1_F1_idmapped.featureXML index 0378223ac..f4a4ed4e0 100644 --- a/src/data/BSA1_F1_idmapped.featureXML +++ b/src/data/BSA1_F1_idmapped.featureXML @@ -1,8476 +1,8476 @@ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 1942.60008303114 - 395.239277484387 - 1.57572e+08 - 0 - 0 - 0.808494 - 2 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 1749.13833503866 - 443.711224390254 - 5.40693e+07 - 0 - 0 - 0.893553 - 2 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 1850.05890690118 - 325.49117966032 - 5.2717e+07 - 0 - 0 - 0.788273 - 3 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 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0.762523 + 1 + + + + + + + + + + + + + + + + + + + + + 1799.99397448666 + 319.487399600059 + 72916.5 + 0 + 0 + 0.774251 + 3 + + + + + + + + + + + + + + + + + + + + + + + + + + + 1798.73709024754 + 481.217046424739 + 72350.1 + 0 + 0 + 0.854945 + 2 + + + + + + + + + + + + + + + + + + + + + 1648.49378695204 + 569.520791440652 + 65775.5 + 0 + 0 + 0.705992 + 3 + + + + + + + + + + + + + + + + + + + + + + + + + + + 1865.54631697755 + 311.196655627092 + 65458.3 + 0 + 0 + 0.892754 + 1 + + + + + + + + + + + + + + + + + + + + + 1650.46662313908 + 330.264019086177 + 60136.1 + 0 + 0 + 0.865523 + 1 + + + + + + + + + + + + + + + + + + + + + 1673.58482514647 + 338.495600192177 + 57300.3 + 0 + 0 + 0.832399 + 1 + + + + + + + + + + + + + + + + + + + + + 1593.86959007379 + 508.247438492578 + 57241.9 + 0 + 0 + 0.72796 + 2 + + + + + + + + + + + + + + + + + + + + + 1722.9424840488 + 545.175730987009 + 57052.5 + 0 + 0 + 0.732768 + 3 + + + + + + + + + + + + + + + + + + + + + 1673.13116181062 + 329.159602803467 + 50060.5 + 0 + 0 + 0.871315 + 1 + + + + + + + + + + + + + + + + + + + + + 1726.24061234872 + 302.196293668864 + 43022.5 + 0 + 0 + 0.786973 + 1 + + + + + + + + + + + + + + + + + + + + + 1926.34240727199 + 437.228633270123 + 42100.6 + 0 + 0 + 0.833002 + 2 + + + + + + + + + + + + + + + + + + + + + 1854.3617392797 + 325.490895560088 + 35323.6 + 0 + 0 + 0.811533 + 3 + + + + + + + + + + + + + + + + + + + + + 1786.66309376502 + 530.275197758271 + 34863 + 0 + 0 + 0.745689 + 1 + + + + + + + + + + + + + + + + + + + + + 1761.9177786468 + 478.719570701758 + 33494.1 + 0 + 0 + 0.822594 + 2 + + + + + + + + + + + + + + + + + + + + + 1752.44169991284 + 316.211986804804 + 26101.4 + 0 + 0 + 0.871625 + 1 + + + + + + + + + + + + + + + + + + + + + From 93bd83422a104a5cd455b44c7b13323cb504fc28 Mon Sep 17 00:00:00 2001 From: "chris.bielow@fu-berlin.de" Date: Tue, 29 Apr 2025 10:11:36 +0200 Subject: [PATCH 2/5] remove UTF8 chars from text --- docs/source/user_guide/feature_linking.rst | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/source/user_guide/feature_linking.rst b/docs/source/user_guide/feature_linking.rst index 34cdbcca8..1b55ef100 100644 --- a/docs/source/user_guide/feature_linking.rst +++ b/docs/source/user_guide/feature_linking.rst @@ -55,7 +55,7 @@ This enables comparisons of peptide signals across runs and is a prerequisite fo Thus, all features across all :py:class:`~.FeatureMap` objects will be combined into a :py:class:`~.ConsensusMap`. Here we use :py:class:`~.FeatureGroupingAlgorithmQT` to find corresponding features with similar RT and m/z across feature maps. If everything goes well, we will have a lot of triplets in our test run (we have have three input maps; one feature from each feature map) within each consensus feature. -Realistically, some features will remain unmatched, forming singletons or doublets—consensus features of size 1 or 2. These undersized consensus features can arise due to several factors. In some cases, the signal may be genuinely absent in the dataset, preventing feature detection. Alternatively, the feature-finding algorithm might fail to identify the feature in all available maps, leading to incomplete linkage. Additionally, suboptimal parameters for feature linking—such as excessive retention time (RT) variations across samples—can contribute to mismatches, further reducing the likelihood of forming larger consensus features. The latter can be corrected for using a process called map alignment, which we adressed in a `previous chapter `_. +Realistically, some features will remain unmatched, forming singletons or doublets (consensus features of size 1 or 2). These undersized consensus features can arise due to several factors. In some cases, the signal may be genuinely absent in the dataset, preventing feature detection. Alternatively, the feature-finding algorithm might fail to identify the feature in all available maps, leading to incomplete linkage. Additionally, suboptimal parameters for feature linking, such as excessive retention time (RT) variations across samples, can contribute to mismatches, further reducing the likelihood of forming larger consensus features. The latter can be corrected for using a process called map alignment, which we adressed in a `previous chapter `_. .. code-block:: python From 44ff3eadf63f621af2efcdc033c41aa4087c8e1e Mon Sep 17 00:00:00 2001 From: "chris.bielow@fu-berlin.de" Date: Tue, 29 Apr 2025 10:42:15 +0200 Subject: [PATCH 3/5] remove sneaky control character from URL --- docs/source/user_guide/map_alignment.rst | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/source/user_guide/map_alignment.rst b/docs/source/user_guide/map_alignment.rst index 99f00a01a..c87c5a356 100644 --- a/docs/source/user_guide/map_alignment.rst +++ b/docs/source/user_guide/map_alignment.rst @@ -78,7 +78,7 @@ Download Example Data # we use featureXML files which already contain PSMs (as obtained by oms.IDMapper()) # ... so we can use all aligners pyOpenMS has to offer feature_files = [ - "BSA1_F1_idmapped.featureXML", + "BSA1_F1_idmapped.featureXML", "BSA2_F1_idmapped.featureXML", "BSA3_F1_idmapped.featureXML", ] From 56e9631de019492f42cd3236f0c4ed752694f9ba Mon Sep 17 00:00:00 2001 From: "chris.bielow@fu-berlin.de" Date: Tue, 29 Apr 2025 10:47:38 +0200 Subject: [PATCH 4/5] fix typos --- docs/source/user_guide/feature_linking.rst | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/source/user_guide/feature_linking.rst b/docs/source/user_guide/feature_linking.rst index 1b55ef100..e94e58ab7 100644 --- a/docs/source/user_guide/feature_linking.rst +++ b/docs/source/user_guide/feature_linking.rst @@ -54,8 +54,8 @@ Feature linking is the process which connects (links) features with similar RT a This enables comparisons of peptide signals across runs and is a prerequisite for downstream statistical analysis. Thus, all features across all :py:class:`~.FeatureMap` objects will be combined into a :py:class:`~.ConsensusMap`. Here we use :py:class:`~.FeatureGroupingAlgorithmQT` to find corresponding features with similar RT and m/z across feature maps. -If everything goes well, we will have a lot of triplets in our test run (we have have three input maps; one feature from each feature map) within each consensus feature. -Realistically, some features will remain unmatched, forming singletons or doublets (consensus features of size 1 or 2). These undersized consensus features can arise due to several factors. In some cases, the signal may be genuinely absent in the dataset, preventing feature detection. Alternatively, the feature-finding algorithm might fail to identify the feature in all available maps, leading to incomplete linkage. Additionally, suboptimal parameters for feature linking, such as excessive retention time (RT) variations across samples, can contribute to mismatches, further reducing the likelihood of forming larger consensus features. The latter can be corrected for using a process called map alignment, which we adressed in a `previous chapter `_. +If everything goes well, we will have a lot of triplets in our test run (we have three input maps; one feature from each feature map) within each consensus feature. +Realistically, some features will remain unmatched, forming singletons or doublets (consensus features of size 1 or 2). These undersized consensus features can arise due to several factors. In some cases, the signal may be genuinely absent in the dataset, preventing feature detection. Alternatively, the feature-finding algorithm might fail to identify the feature in all available maps, leading to incomplete linkage. Additionally, suboptimal parameters for feature linking, such as excessive retention time (RT) variations across samples, can contribute to mismatches, further reducing the likelihood of forming larger consensus features. The latter can be corrected for using a process called map alignment, which we addressed in a `previous chapter `_. .. code-block:: python @@ -65,7 +65,7 @@ Realistically, some features will remain unmatched, forming singletons or double # populate the actual cmap using FeatureGroupingAlgorithmQT feature_grouper = oms.FeatureGroupingAlgorithmQT() - # this is where the work happens: + # execute feature linking: feature_grouper.group(feature_maps, consensus_map) That's it! From 5a56a8475f6b1be109843676072d48782409b1fc Mon Sep 17 00:00:00 2001 From: "chris.bielow@fu-berlin.de" Date: Tue, 29 Apr 2025 10:54:41 +0200 Subject: [PATCH 5/5] more formal text --- docs/source/user_guide/map_alignment.rst | 9 ++++----- 1 file changed, 4 insertions(+), 5 deletions(-) diff --git a/docs/source/user_guide/map_alignment.rst b/docs/source/user_guide/map_alignment.rst index c87c5a356..6094d16ef 100644 --- a/docs/source/user_guide/map_alignment.rst +++ b/docs/source/user_guide/map_alignment.rst @@ -2,10 +2,9 @@ Map Alignment =============== The pyOpenMS map alignment algorithms transform different maps (:term:`peak maps`, :term:`feature maps`) to a common retention time axis. -This is useful, since chromatographic columns are usually less stable, i.e. the RT of identical compounds will vary when repeating the -experiment or running another sample. -We want to correct for this, usually by finding common landmarks across runs and adjusting the RT of all features to minimize the distance -of landmarks across feature maps. +Because chromatographic columns are less stable, retention times of identical compounds can vary across runs. +To correct for RT drift, map alignment attempts to find common landmarks across runs and adjusts the RT of all features to minimize the distance +of landmarks across feature maps (runs). .. image:: img/map_alignment_illustration.png @@ -276,6 +275,6 @@ We can also inspect the underlying transformation and the landmarks it is based # Example usage: plot_transformed_rt_with_trafo(feature_maps, trafos) -While we do not know the underlying features using just this information, we can see that features in corresponding maps are now nicely aligned horizontally (the new RT). Also, it seems a linear model nicely fits all the data points in each feature map. Using a more flexible model like LOWESS will likely result in a worse alignment (try and see for yourself). +While we do not know the underlying features using just this information, we can see that features in corresponding maps are now nicely aligned horizontally (the new RT). Also, it seems a linear model nicely fits all the data points in each feature map. More flexible models (e.g., LOWESS) may overfit and degrade performance—feel; free to experiment. .. image:: img/map_alignment_trafos.png \ No newline at end of file