private DataPointAquisitionResults GetDataAcquisitionResults(MsDataFile myMsDataFile, string currentDataFile, List <Modification> variableModifications, List <Modification> fixedModifications, List <Protein> proteinList, string taskId, CommonParameters combinedParameters, Tolerance initPrecTol, Tolerance initProdTol)
{
var fileNameWithoutExtension = Path.GetFileNameWithoutExtension(currentDataFile);
MassDiffAcceptor searchMode = initPrecTol is PpmTolerance ?
(MassDiffAcceptor) new SinglePpmAroundZeroSearchMode(initPrecTol.Value) :
new SingleAbsoluteAroundZeroSearchMode(initPrecTol.Value);
var listOfSortedms2Scans = GetMs2Scans(myMsDataFile, currentDataFile, combinedParameters).OrderBy(b => b.PrecursorMass).ToArray();
PeptideSpectralMatch[] allPsmsArray = new PeptideSpectralMatch[listOfSortedms2Scans.Length];
Log("Searching with searchMode: " + searchMode, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
Log("Searching with productMassTolerance: " + initProdTol, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
new ClassicSearchEngine(allPsmsArray, listOfSortedms2Scans, variableModifications, fixedModifications, null, null, null, proteinList, searchMode, combinedParameters, this.FileSpecificParameters, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> allPsms = allPsmsArray.Where(b => b != null).ToList();
allPsms = allPsms.OrderByDescending(b => b.Score)
.ThenBy(b => b.PeptideMonisotopicMass.HasValue ? Math.Abs(b.ScanPrecursorMass - b.PeptideMonisotopicMass.Value) : double.MaxValue)
.GroupBy(b => (b.FullFilePath, b.ScanNumber, b.PeptideMonisotopicMass)).Select(b => b.First()).ToList();
new FdrAnalysisEngine(allPsms, searchMode.NumNotches, CommonParameters, this.FileSpecificParameters, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> goodIdentifications = allPsms.Where(b => b.FdrInfo.QValueNotch < 0.001 && !b.IsDecoy && b.FullSequence != null).ToList();
if (!goodIdentifications.Any())
{
return(new DataPointAquisitionResults(null, new List <PeptideSpectralMatch>(), new List <LabeledDataPoint>(), new List <LabeledDataPoint>(), 0, 0, 0, 0));
}
DataPointAquisitionResults currentResult = (DataPointAquisitionResults) new DataPointAcquisitionEngine(
goodIdentifications,
myMsDataFile,
initPrecTol,
CalibrationParameters.MinMS1IsotopicPeaksNeededForConfirmedIdentification,
CommonParameters,
this.FileSpecificParameters,
new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
return(currentResult);
}
protected override MyTaskResults RunSpecific(string OutputFolder, List <DbForTask> dbFilenameList, List <string> currentRawFileList, string taskId, FileSpecificParameters[] fileSettingsList)
{
LoadModifications(taskId, out var variableModifications, out var fixedModifications, out var localizeableModificationTypes);
// load proteins
List <Protein> proteinList = LoadProteins(taskId, dbFilenameList, true, DecoyType.Reverse, localizeableModificationTypes, CommonParameters);
// write prose settings
ProseCreatedWhileRunning.Append("The following calibration settings were used: ");
ProseCreatedWhileRunning.Append("protease = " + CommonParameters.DigestionParams.Protease + "; ");
ProseCreatedWhileRunning.Append("maximum missed cleavages = " + CommonParameters.DigestionParams.MaxMissedCleavages + "; ");
ProseCreatedWhileRunning.Append("minimum peptide length = " + CommonParameters.DigestionParams.MinPeptideLength + "; ");
ProseCreatedWhileRunning.Append(CommonParameters.DigestionParams.MaxPeptideLength == int.MaxValue ?
"maximum peptide length = unspecified; " :
"maximum peptide length = " + CommonParameters.DigestionParams.MaxPeptideLength + "; ");
ProseCreatedWhileRunning.Append("initiator methionine behavior = " + CommonParameters.DigestionParams.InitiatorMethionineBehavior + "; ");
ProseCreatedWhileRunning.Append("fixed modifications = " + string.Join(", ", fixedModifications.Select(m => m.IdWithMotif)) + "; ");
ProseCreatedWhileRunning.Append("variable modifications = " + string.Join(", ", variableModifications.Select(m => m.IdWithMotif)) + "; ");
ProseCreatedWhileRunning.Append("max mods per peptide = " + CommonParameters.DigestionParams.MaxModsForPeptide + "; ");
ProseCreatedWhileRunning.Append("max modification isoforms = " + CommonParameters.DigestionParams.MaxModificationIsoforms + "; ");
ProseCreatedWhileRunning.Append("precursor mass tolerance = " + CommonParameters.PrecursorMassTolerance + "; ");
ProseCreatedWhileRunning.Append("product mass tolerance = " + CommonParameters.ProductMassTolerance + ". ");
ProseCreatedWhileRunning.Append("The combined search database contained " + proteinList.Count(p => !p.IsDecoy) + " non-decoy protein entries including " + proteinList.Count(p => p.IsContaminant) + " contaminant sequences. ");
// start the calibration task
Status("Calibrating...", new List <string> {
taskId
});
MyTaskResults = new MyTaskResults(this)
{
NewSpectra = new List <string>(),
NewFileSpecificTomls = new List <string>()
};
var myFileManager = new MyFileManager(true);
List <string> spectraFilesAfterCalibration = new List <string>();
for (int spectraFileIndex = 0; spectraFileIndex < currentRawFileList.Count; spectraFileIndex++)
{
if (GlobalVariables.StopLoops)
{
break;
}
bool couldNotFindEnoughDatapoints = false;
// get filename stuff
var originalUncalibratedFilePath = currentRawFileList[spectraFileIndex];
var originalUncalibratedFilenameWithoutExtension = Path.GetFileNameWithoutExtension(originalUncalibratedFilePath);
string calibratedFilePath = Path.Combine(OutputFolder, originalUncalibratedFilenameWithoutExtension + CalibSuffix + ".mzML");
// mark the file as in-progress
StartingDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
CommonParameters combinedParams = SetAllFileSpecificCommonParams(CommonParameters, fileSettingsList[spectraFileIndex]);
// load the file
Status("Loading spectra file...", new List <string> {
taskId, "Individual Spectra Files"
});
var myMsDataFile = myFileManager.LoadFile(originalUncalibratedFilePath, CommonParameters);
// get datapoints to fit calibration function to
Status("Acquiring calibration data points...", new List <string> {
taskId, "Individual Spectra Files"
});
DataPointAquisitionResults acquisitionResults = null;
for (int i = 1; i <= 5; i++)
{
acquisitionResults = GetDataAcquisitionResults(myMsDataFile, originalUncalibratedFilePath, variableModifications, fixedModifications, proteinList, taskId, combinedParams, combinedParams.PrecursorMassTolerance, combinedParams.ProductMassTolerance);
// enough data points to calibrate?
if (acquisitionResults.Psms.Count >= NumRequiredPsms && acquisitionResults.Ms1List.Count > NumRequiredMs1Datapoints && acquisitionResults.Ms2List.Count > NumRequiredMs2Datapoints)
{
break;
}
if (i == 1) // failed round 1
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(20);
CommonParameters.ProductMassTolerance = new PpmTolerance(50);
}
else if (i == 2) // failed round 2
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(30);
CommonParameters.ProductMassTolerance = new PpmTolerance(100);
}
else if (i == 3) // failed round 3
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(40);
CommonParameters.ProductMassTolerance = new PpmTolerance(150);
}
else // failed round 4
{
if (acquisitionResults.Psms.Count < NumRequiredPsms)
{
Warn("Calibration failure! Could not find enough high-quality PSMs. Required " + NumRequiredPsms + ", saw " + acquisitionResults.Psms.Count);
}
if (acquisitionResults.Ms1List.Count < NumRequiredMs1Datapoints)
{
Warn("Calibration failure! Could not find enough MS1 datapoints. Required " + NumRequiredMs1Datapoints + ", saw " + acquisitionResults.Ms1List.Count);
}
if (acquisitionResults.Ms2List.Count < NumRequiredMs2Datapoints)
{
Warn("Calibration failure! Could not find enough MS2 datapoints. Required " + NumRequiredMs2Datapoints + ", saw " + acquisitionResults.Ms2List.Count);
}
couldNotFindEnoughDatapoints = true;
FinishedDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
break;
}
Warn("Could not find enough PSMs to calibrate with; opening up tolerances to " +
Math.Round(CommonParameters.PrecursorMassTolerance.Value, 2) + " ppm precursor and " +
Math.Round(CommonParameters.ProductMassTolerance.Value, 2) + " ppm product");
}
if (couldNotFindEnoughDatapoints)
{
spectraFilesAfterCalibration.Add(Path.GetFileNameWithoutExtension(currentRawFileList[spectraFileIndex]));
ReportProgress(new ProgressEventArgs(100, "Failed to calibrate!", new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
}));
continue;
}
// stats before calibration
int prevPsmCount = acquisitionResults.Psms.Count;
double preCalibrationPrecursorErrorIqr = acquisitionResults.PsmPrecursorIqrPpmError;
double preCalibrationProductErrorIqr = acquisitionResults.PsmProductIqrPpmError;
// generate calibration function and shift data points
Status("Calibrating...", new List <string> {
taskId, "Individual Spectra Files"
});
CalibrationEngine engine = new CalibrationEngine(myMsDataFile, acquisitionResults, CommonParameters, FileSpecificParameters, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
engine.Run();
//update file
myMsDataFile = engine.CalibratedDataFile;
// do another search to evaluate calibration results
Status("Post-calibration search...", new List <string> {
taskId, "Individual Spectra Files"
});
acquisitionResults = GetDataAcquisitionResults(myMsDataFile, originalUncalibratedFilePath, variableModifications, fixedModifications, proteinList, taskId, combinedParams, combinedParams.PrecursorMassTolerance, combinedParams.ProductMassTolerance);
//generate calibration function and shift data points AGAIN because it's fast and contributes new data
Status("Calibrating...", new List <string> {
taskId, "Individual Spectra Files"
});
engine = new CalibrationEngine(myMsDataFile, acquisitionResults, CommonParameters, FileSpecificParameters, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
engine.Run();
//update file
myMsDataFile = engine.CalibratedDataFile;
// write the calibrated mzML file
MzmlMethods.CreateAndWriteMyMzmlWithCalibratedSpectra(myMsDataFile, calibratedFilePath, false);
myFileManager.DoneWithFile(originalUncalibratedFilePath);
// stats after calibration
int postCalibrationPsmCount = acquisitionResults.Psms.Count;
double postCalibrationPrecursorErrorIqr = acquisitionResults.PsmPrecursorIqrPpmError;
double postCalibrationProductErrorIqr = acquisitionResults.PsmProductIqrPpmError;
// did the data improve? (not used for anything yet...)
bool improvement = ImprovGlobal(preCalibrationPrecursorErrorIqr, preCalibrationProductErrorIqr, prevPsmCount, postCalibrationPsmCount, postCalibrationPrecursorErrorIqr, postCalibrationProductErrorIqr);
// write toml settings for the calibrated file
var newTomlFileName = Path.Combine(OutputFolder, originalUncalibratedFilenameWithoutExtension + CalibSuffix + ".toml");
var fileSpecificParams = new FileSpecificParameters();
// carry over file-specific parameters from the uncalibrated file to the calibrated one
if (fileSettingsList[spectraFileIndex] != null)
{
fileSpecificParams = fileSettingsList[spectraFileIndex].Clone();
}
//suggest 4 * interquartile range as the ppm tolerance
fileSpecificParams.PrecursorMassTolerance = new PpmTolerance((4.0 * postCalibrationPrecursorErrorIqr) + Math.Abs(acquisitionResults.PsmPrecursorMedianPpmError));
fileSpecificParams.ProductMassTolerance = new PpmTolerance((4.0 * postCalibrationProductErrorIqr) + Math.Abs(acquisitionResults.PsmProductMedianPpmError));
Toml.WriteFile(fileSpecificParams, newTomlFileName, tomlConfig);
FinishedWritingFile(newTomlFileName, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
// finished calibrating this file
spectraFilesAfterCalibration.Add(Path.GetFileNameWithoutExtension(calibratedFilePath));
FinishedWritingFile(calibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
MyTaskResults.NewSpectra.Add(calibratedFilePath);
MyTaskResults.NewFileSpecificTomls.Add(newTomlFileName);
FinishedDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
ReportProgress(new ProgressEventArgs(100, "Done!", new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
}));
}
// re-write experimental design (if it has been defined) with new calibrated file names
string assumedPathToExperDesign = Directory.GetParent(currentRawFileList.First()).FullName;
assumedPathToExperDesign = Path.Combine(assumedPathToExperDesign, GlobalVariables.ExperimentalDesignFileName);
if (File.Exists(assumedPathToExperDesign))
{
WriteNewExperimentalDesignFile(assumedPathToExperDesign, OutputFolder, spectraFilesAfterCalibration);
}
// finished calibrating all files for the task
ReportProgress(new ProgressEventArgs(100, "Done!", new List <string> {
taskId, "Individual Spectra Files"
}));
return(MyTaskResults);
}
protected override MyTaskResults RunSpecific(string OutputFolder, List <DbForTask> dbFilenameList, List <string> currentRawFileList, string taskId, FileSpecificParameters[] fileSettingsList)
{
// load modifications
Status("Loading modifications...", new List <string> {
taskId
});
List <ModificationWithMass> variableModifications = GlobalVariables.AllModsKnown.OfType <ModificationWithMass>().Where(b => CommonParameters.ListOfModsVariable.Contains((b.modificationType, b.id))).ToList();
List <ModificationWithMass> fixedModifications = GlobalVariables.AllModsKnown.OfType <ModificationWithMass>().Where(b => CommonParameters.ListOfModsFixed.Contains((b.modificationType, b.id))).ToList();
List <string> localizeableModificationTypes = GlobalVariables.AllModTypesKnown.ToList();
// what types of fragment ions to search for
List <ProductType> ionTypes = new List <ProductType>();
if (CommonParameters.BIons)
{
ionTypes.Add(ProductType.BnoB1ions);
}
if (CommonParameters.YIons)
{
ionTypes.Add(ProductType.Y);
}
if (CommonParameters.ZdotIons)
{
ionTypes.Add(ProductType.Zdot);
}
if (CommonParameters.CIons)
{
ionTypes.Add(ProductType.C);
}
// load proteins
List <Protein> proteinList = LoadProteins(taskId, dbFilenameList, true, DecoyType.Reverse, localizeableModificationTypes, CommonParameters);
// write prose settings
ProseCreatedWhileRunning.Append("The following calibration settings were used: ");
ProseCreatedWhileRunning.Append("protease = " + CommonParameters.DigestionParams.Protease + "; ");
ProseCreatedWhileRunning.Append("maximum missed cleavages = " + CommonParameters.DigestionParams.MaxMissedCleavages + "; ");
ProseCreatedWhileRunning.Append("minimum peptide length = " + CommonParameters.DigestionParams.MinPeptideLength + "; ");
ProseCreatedWhileRunning.Append(CommonParameters.DigestionParams.MaxPeptideLength == int.MaxValue ?
"maximum peptide length = unspecified; " :
"maximum peptide length = " + CommonParameters.DigestionParams.MaxPeptideLength + "; ");
ProseCreatedWhileRunning.Append("initiator methionine behavior = " + CommonParameters.DigestionParams.InitiatorMethionineBehavior + "; ");
ProseCreatedWhileRunning.Append("fixed modifications = " + string.Join(", ", fixedModifications.Select(m => m.id)) + "; ");
ProseCreatedWhileRunning.Append("variable modifications = " + string.Join(", ", variableModifications.Select(m => m.id)) + "; ");
ProseCreatedWhileRunning.Append("max mods per peptide = " + CommonParameters.DigestionParams.MaxModsForPeptide + "; ");
ProseCreatedWhileRunning.Append("max modification isoforms = " + CommonParameters.DigestionParams.MaxModificationIsoforms + "; ");
ProseCreatedWhileRunning.Append("precursor mass tolerance = " + CommonParameters.PrecursorMassTolerance + "; ");
ProseCreatedWhileRunning.Append("product mass tolerance = " + CommonParameters.ProductMassTolerance + ". ");
ProseCreatedWhileRunning.Append("The combined search database contained " + proteinList.Count(p => !p.IsDecoy) + " non-decoy protein entries including " + proteinList.Count(p => p.IsContaminant) + " contaminant sequences. ");
// start the calibration task
Status("Calibrating...", new List <string> {
taskId
});
MyTaskResults = new MyTaskResults(this)
{
NewSpectra = new List <string>(),
NewFileSpecificTomls = new List <string>()
};
object lock1 = new object();
var myFileManager = new MyFileManager(true);
for (int spectraFileIndex = 0; spectraFileIndex < currentRawFileList.Count; spectraFileIndex++)
{
if (GlobalVariables.StopLoops)
{
break;
}
// get filename stuff
var originalUncalibratedFilePath = currentRawFileList[spectraFileIndex];
var originalUncalibratedFilenameWithoutExtension = Path.GetFileNameWithoutExtension(originalUncalibratedFilePath);
string calibratedFilePath = Path.Combine(OutputFolder, originalUncalibratedFilenameWithoutExtension + CalibSuffix + ".mzML");
// mark the file as in-progress
StartingDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
CommonParameters combinedParams = SetAllFileSpecificCommonParams(CommonParameters, fileSettingsList[spectraFileIndex]);
MsDataFile myMsDataFile;
// load the file
Status("Loading spectra file...", new List <string> {
taskId, "Individual Spectra Files"
});
lock (lock1)
{
myMsDataFile = myFileManager.LoadFile(originalUncalibratedFilePath, CommonParameters.TopNpeaks, CommonParameters.MinRatio, CommonParameters.TrimMs1Peaks, CommonParameters.TrimMsMsPeaks, CommonParameters);
}
// get datapoints to fit calibration function to
Status("Acquiring calibration data points...", new List <string> {
taskId, "Individual Spectra Files"
});
DataPointAquisitionResults acquisitionResults = null;
for (int i = 1; i <= 5; i++)
{
acquisitionResults = GetDataAcquisitionResults(myMsDataFile, originalUncalibratedFilePath, variableModifications, fixedModifications, proteinList, taskId, combinedParams, combinedParams.PrecursorMassTolerance, combinedParams.ProductMassTolerance);
// enough data points to calibrate?
if (acquisitionResults.Psms.Count >= NumRequiredPsms && acquisitionResults.Ms1List.Count > NumRequiredMs1Datapoints && acquisitionResults.Ms2List.Count > NumRequiredMs2Datapoints)
{
break;
}
if (i == 1) // failed round 1
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(20);
CommonParameters.ProductMassTolerance = new PpmTolerance(50);
}
else if (i == 2) // failed round 2
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(30);
CommonParameters.ProductMassTolerance = new PpmTolerance(100);
}
else if (i == 3) // failed round 3
{
CommonParameters.PrecursorMassTolerance = new PpmTolerance(40);
CommonParameters.ProductMassTolerance = new PpmTolerance(150);
}
else // failed round 4
{
if (acquisitionResults.Psms.Count < NumRequiredPsms)
{
Warn("Calibration failure! Could not find enough high-quality PSMs. Required " + NumRequiredPsms + ", saw " + acquisitionResults.Psms.Count);
}
if (acquisitionResults.Ms1List.Count < NumRequiredMs1Datapoints)
{
Warn("Calibration failure! Could not find enough MS1 datapoints. Required " + NumRequiredMs1Datapoints + ", saw " + acquisitionResults.Ms1List.Count);
}
if (acquisitionResults.Ms2List.Count < NumRequiredMs2Datapoints)
{
Warn("Calibration failure! Could not find enough MS2 datapoints. Required " + NumRequiredMs2Datapoints + ", saw " + acquisitionResults.Ms2List.Count);
}
FinishedDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
return(MyTaskResults);
}
Warn("Could not find enough PSMs to calibrate with; opening up tolerances to " +
Math.Round(CommonParameters.PrecursorMassTolerance.Value, 2) + " ppm precursor and " +
Math.Round(CommonParameters.ProductMassTolerance.Value, 2) + " ppm product");
}
// stats before calibration
int prevPsmCount = acquisitionResults.Psms.Count;
double preCalibrationPrecursorErrorIqr = acquisitionResults.PsmPrecursorIqrPpmError;
double preCalibrationProductErrorIqr = acquisitionResults.PsmProductIqrPpmError;
// generate calibration function and shift data points
Status("Calibrating...", new List <string> {
taskId, "Individual Spectra Files"
});
new CalibrationEngine(myMsDataFile, acquisitionResults, CommonParameters, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
}).Run();
// do another search to evaluate calibration results
Status("Post-calibration search...", new List <string> {
taskId, "Individual Spectra Files"
});
acquisitionResults = GetDataAcquisitionResults(myMsDataFile, originalUncalibratedFilePath, variableModifications, fixedModifications, proteinList, taskId, combinedParams, combinedParams.PrecursorMassTolerance, combinedParams.ProductMassTolerance);
// stats after calibration
int postCalibrationPsmCount = acquisitionResults.Psms.Count;
double postCalibrationPrecursorErrorIqr = acquisitionResults.PsmPrecursorIqrPpmError;
double postCalibrationProductErrorIqr = acquisitionResults.PsmProductIqrPpmError;
// did the data improve? (not used for anything yet...)
bool improvement = ImprovGlobal(preCalibrationPrecursorErrorIqr, preCalibrationProductErrorIqr, prevPsmCount, postCalibrationPsmCount, postCalibrationPrecursorErrorIqr, postCalibrationProductErrorIqr);
// write toml settings for the calibrated file
var newTomlFileName = Path.Combine(OutputFolder, originalUncalibratedFilenameWithoutExtension + CalibSuffix + ".toml");
var fileSpecificParams = new FileSpecificParameters();
// carry over file-specific parameters from the uncalibrated file to the calibrated one
if (fileSettingsList[spectraFileIndex] != null)
{
fileSpecificParams = fileSettingsList[spectraFileIndex].Clone();
}
// don't write over ppm tolerances if they've been specified by the user already in the file-specific settings
// otherwise, suggest 4 * interquartile range as the ppm tolerance
if (fileSpecificParams.PrecursorMassTolerance == null)
{
fileSpecificParams.PrecursorMassTolerance = new PpmTolerance((4.0 * postCalibrationPrecursorErrorIqr) + Math.Abs(acquisitionResults.PsmPrecursorMedianPpmError));
}
if (fileSpecificParams.ProductMassTolerance == null)
{
fileSpecificParams.ProductMassTolerance = new PpmTolerance((4.0 * postCalibrationProductErrorIqr) + Math.Abs(acquisitionResults.PsmProductMedianPpmError));
}
Toml.WriteFile(fileSpecificParams, newTomlFileName, tomlConfig);
FinishedWritingFile(newTomlFileName, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
// write the calibrated mzML file
MzmlMethods.CreateAndWriteMyMzmlWithCalibratedSpectra(myMsDataFile, calibratedFilePath, false);
myFileManager.DoneWithFile(originalUncalibratedFilePath);
// finished calibrating this file
FinishedWritingFile(calibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
});
MyTaskResults.NewSpectra.Add(calibratedFilePath);
MyTaskResults.NewFileSpecificTomls.Add(newTomlFileName);
FinishedDataFile(originalUncalibratedFilePath, new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilePath
});
ReportProgress(new ProgressEventArgs(100, "Done!", new List <string> {
taskId, "Individual Spectra Files", originalUncalibratedFilenameWithoutExtension
}));
}
// re-write experimental design (if it has been defined) with new calibrated file names
string assumedPathToExperDesign = Directory.GetParent(currentRawFileList.First()).FullName;
assumedPathToExperDesign = Path.Combine(assumedPathToExperDesign, GlobalVariables.ExperimentalDesignFileName);
List <string> newExperimentalDesignOutput = new List <string>();
if (File.Exists(assumedPathToExperDesign))
{
var lines = File.ReadAllLines(assumedPathToExperDesign);
for (int i = 0; i < lines.Length; i++)
{
// header of experimental design file
if (i == 0)
{
newExperimentalDesignOutput.Add(lines[i]);
}
else
{
var split = lines[i].Split('\t');
string newline = Path.GetFileNameWithoutExtension(split[0]) + CalibSuffix + "\t";
for (int j = 1; j < split.Length; j++)
{
newline += split[j] + "\t";
}
newExperimentalDesignOutput.Add(newline);
}
}
}
File.WriteAllLines(Path.Combine(OutputFolder, GlobalVariables.ExperimentalDesignFileName), newExperimentalDesignOutput);
// finished calibrating all files for the task
ReportProgress(new ProgressEventArgs(100, "Done!", new List <string> {
taskId, "Individual Spectra Files"
}));
return(MyTaskResults);
}
private DataPointAquisitionResults GetDataAcquisitionResults(MsDataFile myMsDataFile, string currentDataFile, List <ModificationWithMass> variableModifications, List <ModificationWithMass> fixedModifications, List <Protein> proteinList, string taskId, CommonParameters combinedParameters, Tolerance initPrecTol, Tolerance initProdTol)
{
var fileNameWithoutExtension = Path.GetFileNameWithoutExtension(currentDataFile);
MassDiffAcceptor searchMode = initPrecTol is PpmTolerance ?
(MassDiffAcceptor) new SinglePpmAroundZeroSearchMode(initPrecTol.Value) :
new SingleAbsoluteAroundZeroSearchMode(initPrecTol.Value);
FragmentTypes fragmentTypesForCalibration = FragmentTypes.None;
if (combinedParameters.BIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.b;
}
if (combinedParameters.YIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.y;
}
if (combinedParameters.CIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.c;
}
if (combinedParameters.ZdotIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.zdot;
}
var listOfSortedms2Scans = GetMs2Scans(myMsDataFile, currentDataFile, combinedParameters.DoPrecursorDeconvolution, combinedParameters.UseProvidedPrecursorInfo, combinedParameters.DeconvolutionIntensityRatio, combinedParameters.DeconvolutionMaxAssumedChargeState, combinedParameters.DeconvolutionMassTolerance).OrderBy(b => b.PrecursorMass).ToArray();
PeptideSpectralMatch[] allPsmsArray = new PeptideSpectralMatch[listOfSortedms2Scans.Length];
List <ProductType> productTypes = new List <ProductType>();
if (combinedParameters.BIons)
{
productTypes.Add(ProductType.B);
}
if (combinedParameters.YIons)
{
productTypes.Add(ProductType.Y);
}
if (combinedParameters.CIons)
{
productTypes.Add(ProductType.C);
}
if (combinedParameters.ZdotIons)
{
productTypes.Add(ProductType.Zdot);
}
Log("Searching with searchMode: " + searchMode, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
Log("Searching with productMassTolerance: " + initProdTol, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
new ClassicSearchEngine(allPsmsArray, listOfSortedms2Scans, variableModifications, fixedModifications, proteinList, productTypes, searchMode, combinedParameters, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> allPsms = allPsmsArray.Where(p => p != null).ToList();
var compactPeptideToProteinPeptideMatching = ((SequencesToActualProteinPeptidesEngineResults) new SequencesToActualProteinPeptidesEngine
(allPsms, proteinList, fixedModifications, variableModifications, productTypes, new List <DigestionParams> {
combinedParameters.DigestionParams
},
combinedParameters.ReportAllAmbiguity, combinedParameters, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run()).CompactPeptideToProteinPeptideMatching;
foreach (var huh in allPsms)
{
huh.MatchToProteinLinkedPeptides(compactPeptideToProteinPeptideMatching);
}
allPsms = allPsms.OrderByDescending(b => b.Score)
.ThenBy(b => b.PeptideMonisotopicMass.HasValue ? Math.Abs(b.ScanPrecursorMass - b.PeptideMonisotopicMass.Value) : double.MaxValue)
.GroupBy(b => (b.FullFilePath, b.ScanNumber, b.PeptideMonisotopicMass)).Select(b => b.First()).ToList();
new FdrAnalysisEngine(allPsms, searchMode.NumNotches, CommonParameters, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> goodIdentifications = allPsms.Where(b => b.FdrInfo.QValueNotch < 0.01 && !b.IsDecoy && b.FullSequence != null).ToList();
if (!goodIdentifications.Any())
{
return(new DataPointAquisitionResults(null, new List <PeptideSpectralMatch>(), new List <LabeledDataPoint>(), new List <LabeledDataPoint>(), 0, 0, 0, 0));
}
DataPointAquisitionResults currentResult = (DataPointAquisitionResults) new DataPointAcquisitionEngine(
goodIdentifications,
myMsDataFile,
initPrecTol,
initProdTol,
CalibrationParameters.NumFragmentsNeededForEveryIdentification,
CalibrationParameters.MinMS1IsotopicPeaksNeededForConfirmedIdentification,
CalibrationParameters.MinMS2IsotopicPeaksNeededForConfirmedIdentification,
fragmentTypesForCalibration,
CommonParameters,
new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
return(currentResult);
}
private (List <PeptideSpectralMatch>, DataPointAquisitionResults) GetDataAcquisitionResults(IMsDataFile <IMsDataScan <IMzSpectrum <IMzPeak> > > myMsDataFile, string currentDataFile, List <ModificationWithMass> variableModifications, List <ModificationWithMass> fixedModifications, List <Protein> proteinList, string taskId, ICommonParameters combinedParameters, Tolerance initPrecTol, Tolerance initProdTol)
{
var fileNameWithoutExtension = Path.GetFileNameWithoutExtension(currentDataFile);
MassDiffAcceptor searchMode;
if (initPrecTol is PpmTolerance)
{
searchMode = new SinglePpmAroundZeroSearchMode(initPrecTol.Value);
}
else
{
searchMode = new SingleAbsoluteAroundZeroSearchMode(initPrecTol.Value);
}
FragmentTypes fragmentTypesForCalibration = FragmentTypes.None;
if (combinedParameters.BIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.b;
}
if (combinedParameters.YIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.y;
}
if (combinedParameters.CIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.c;
}
if (combinedParameters.ZdotIons)
{
fragmentTypesForCalibration = fragmentTypesForCalibration | FragmentTypes.zdot;
}
var listOfSortedms2Scans = GetMs2Scans(myMsDataFile, currentDataFile, combinedParameters.DoPrecursorDeconvolution, combinedParameters.UseProvidedPrecursorInfo, combinedParameters.DeconvolutionIntensityRatio, combinedParameters.DeconvolutionMaxAssumedChargeState, combinedParameters.DeconvolutionMassTolerance).OrderBy(b => b.PrecursorMass).ToArray();
PeptideSpectralMatch[] allPsmsArray = new PeptideSpectralMatch[listOfSortedms2Scans.Length];
List <ProductType> lp = new List <ProductType>();
if (combinedParameters.BIons)
{
lp.Add(ProductType.B);
}
if (combinedParameters.YIons)
{
lp.Add(ProductType.Y);
}
if (combinedParameters.CIons)
{
lp.Add(ProductType.C);
}
if (combinedParameters.ZdotIons)
{
lp.Add(ProductType.Zdot);
}
Log("Searching with searchMode: " + searchMode, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
Log("Searching with productMassTolerance: " + initProdTol, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
});
new ClassicSearchEngine(allPsmsArray, listOfSortedms2Scans, variableModifications, fixedModifications, proteinList, lp, searchMode, false, combinedParameters, initProdTol, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> allPsms = allPsmsArray.ToList();
Dictionary <CompactPeptideBase, HashSet <PeptideWithSetModifications> > compactPeptideToProteinPeptideMatching = ((SequencesToActualProteinPeptidesEngineResults) new SequencesToActualProteinPeptidesEngine(allPsms, proteinList, fixedModifications, variableModifications, lp, new List <IDigestionParams> {
combinedParameters.DigestionParams
}, combinedParameters.ReportAllAmbiguity, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run()).CompactPeptideToProteinPeptideMatching;
foreach (var huh in allPsms)
{
if (huh != null)
{
huh.MatchToProteinLinkedPeptides(compactPeptideToProteinPeptideMatching);
}
}
allPsms = allPsms.Where(b => b != null).OrderByDescending(b => b.Score).ThenBy(b => b.PeptideMonisotopicMass.HasValue ? Math.Abs(b.ScanPrecursorMass - b.PeptideMonisotopicMass.Value) : double.MaxValue).GroupBy(b => (b.FullFilePath, b.ScanNumber, b.PeptideMonisotopicMass)).Select(b => b.First()).ToList();
new FdrAnalysisEngine(allPsms, searchMode.NumNotches, false, new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
List <PeptideSpectralMatch> goodIdentifications = allPsms.Where(b => b.FdrInfo.QValueNotch < 0.01 && !b.IsDecoy && b.FullSequence != null).ToList();
if (!goodIdentifications.Any())
{
Warn("No PSMs below 1% FDR observed!");
return(new List <PeptideSpectralMatch>(), null);
}
var dissociationTypes = MetaMorpheusEngine.DetermineDissociationType(lp);
foreach (var psm in allPsms)
{
var theScan = myMsDataFile.GetOneBasedScan(psm.ScanNumber);
double thePrecursorMass = psm.ScanPrecursorMass;
foreach (var huh in lp)
{
var ionMasses = psm.CompactPeptides.First().Key.ProductMassesMightHaveDuplicatesAndNaNs(new List <ProductType> {
huh
});
Array.Sort(ionMasses);
List <double> matchedIonMassesList = new List <double>();
List <double> productMassErrorDaList = new List <double>();
List <double> productMassErrorPpmList = new List <double>();
LocalizationEngine.MatchIons(theScan, initProdTol, ionMasses, matchedIonMassesList, productMassErrorDaList, productMassErrorPpmList, thePrecursorMass, dissociationTypes, false);
double[] matchedIonMassesOnlyMatches = matchedIonMassesList.ToArray();
psm.MatchedIonDictOnlyMatches.Add(huh, matchedIonMassesOnlyMatches);
psm.ProductMassErrorDa.Add(huh, productMassErrorDaList.ToArray());
psm.ProductMassErrorPpm.Add(huh, productMassErrorPpmList.ToArray());
}
}
DataPointAquisitionResults currentResult = (DataPointAquisitionResults) new DataPointAcquisitionEngine(
goodIdentifications,
myMsDataFile,
initPrecTol,
initProdTol,
CalibrationParameters.NumFragmentsNeededForEveryIdentification,
CalibrationParameters.MinMS1IsotopicPeaksNeededForConfirmedIdentification,
CalibrationParameters.MinMS2IsotopicPeaksNeededForConfirmedIdentification,
fragmentTypesForCalibration,
new List <string> {
taskId, "Individual Spectra Files", fileNameWithoutExtension
}).Run();
return(goodIdentifications, currentResult);
}
