private int GetScanData(XRawFileIO reader, clsScanInfo scanInfo, out double[] mzList, out double[] intensityList)
{
if (scanInfo.IsFTMS)
{
var labelDataCount = reader.GetScanLabelData(scanInfo.ScanNumber, out var labelData);
if (labelDataCount > 0)
{
mzList = new double[labelDataCount];
intensityList = new double[labelDataCount];
for (var i = 0; i < labelDataCount; i++)
{
mzList[i] = labelData[i].Mass;
intensityList[i] = labelData[i].Intensity;
}
return(labelDataCount);
}
}
var dataPointCount = reader.GetScanData(scanInfo.ScanNumber, out mzList, out intensityList, 0, true);
return(dataPointCount);
}
private List<XYData> LoadRawSpectra(XRawFileIO rawReader, int scan)
{
double[] mz;
double[] intensities;
rawReader.GetScanData(scan, out mz, out intensities);
if (mz == null)
return new List<XYData>();
var data = new List<XYData>(mz.Length);
for (var i = 0; i < mz.Length; i++)
{
var intensity = intensities[i];
data.Add(new XYData(mz[i], intensity));
}
return data;
}
private static void TestReader(string rawFilePath, bool centroid = false, bool testSumming = false, int scanStart = 0, int scanEnd = 0)
{
try
{
var rawFile = ResolveDataFile(rawFilePath);
if (rawFile == null)
{
return;
}
var options = new ThermoReaderOptions {
LoadMSMethodInfo = mLoadMethods
};
using (var reader = new XRawFileIO(rawFile.FullName, options, mTraceMode))
{
RegisterEvents(reader);
var numScans = reader.GetNumScans();
var collisionEnergyList = string.Empty;
ShowMethod(reader);
var scanStep = 1;
if (scanStart < 1)
{
scanStart = 1;
}
if (scanEnd < 1)
{
scanEnd = numScans;
scanStep = 21;
}
else
{
if (scanEnd < scanStart)
{
scanEnd = scanStart;
}
}
if (scanEnd > numScans)
{
scanEnd = numScans;
}
var msLevelStats = new Dictionary <int, int>();
Console.WriteLine();
Console.WriteLine("Reading data for scans {0} to {1}, step {2}", scanStart, scanEnd, scanStep);
for (var scanNum = scanStart; scanNum <= scanEnd; scanNum += scanStep)
{
if (scanNum > reader.ScanEnd)
{
ConsoleMsgUtils.ShowWarning("Exiting for loop since scan number {0} is greater than the max scan number, {1}", scanNum, reader.ScanEnd);
break;
}
if (mOnlyLoadMSLevelInfo)
{
var msLevel = reader.GetMSLevel(scanNum);
if (msLevelStats.TryGetValue(msLevel, out var msLevelObsCount))
{
msLevelStats[msLevel] = msLevelObsCount + 1;
}
else
{
msLevelStats.Add(msLevel, 1);
}
if (mScanInfoInterval <= 0 || scanNum % mScanInfoInterval == 0)
{
Console.WriteLine("Scan " + scanNum);
}
continue;
}
var success = reader.GetScanInfo(scanNum, out clsScanInfo scanInfo);
if (!success)
{
continue;
}
if (mScanInfoInterval <= 0 || scanNum % mScanInfoInterval == 0)
{
Console.WriteLine("Scan " + scanNum + " at " + scanInfo.RetentionTime.ToString("0.00") + " minutes: " + scanInfo.FilterText);
}
if (mLoadCollisionEnergies)
{
var collisionEnergies = reader.GetCollisionEnergy(scanNum);
if (collisionEnergies.Count == 0)
{
collisionEnergyList = string.Empty;
}
else if (collisionEnergies.Count >= 1)
{
collisionEnergyList = collisionEnergies[0].ToString("0.0");
if (collisionEnergies.Count > 1)
{
for (var index = 1; index <= collisionEnergies.Count - 1; index++)
{
collisionEnergyList += ", " + collisionEnergies[index].ToString("0.0");
}
}
}
if (string.IsNullOrEmpty(collisionEnergyList))
{
Console.WriteLine();
}
else
{
Console.WriteLine("; CE " + collisionEnergyList);
}
}
if (mGetScanEvents)
{
if (scanInfo.TryGetScanEvent("Monoisotopic M/Z:", out var monoMZ))
{
Console.WriteLine("Monoisotopic M/Z: " + monoMZ);
}
if (scanInfo.TryGetScanEvent("Charge State", out var chargeState, true))
{
Console.WriteLine("Charge State: " + chargeState);
}
if (scanInfo.TryGetScanEvent("MS2 Isolation Width", out var isolationWidth, true))
{
Console.WriteLine("MS2 Isolation Width: " + isolationWidth);
}
}
if (!mLoadScanData || (scanNum % 50 != 0 && scanEnd - scanStart > 50))
{
continue;
}
// Get the data for scan scanNum
Console.WriteLine();
Console.WriteLine("Spectrum for scan " + scanNum);
reader.GetScanData(scanNum, out var mzList, out var intensityList, 0, centroid);
var mzDisplayStepSize = 50;
if (centroid)
{
mzDisplayStepSize = 1;
}
for (var i = 0; i <= mzList.Length - 1; i += mzDisplayStepSize)
{
Console.WriteLine(" " + mzList[i].ToString("0.000") + " mz " + intensityList[i].ToString("0"));
}
Console.WriteLine();
const int scansToSum = 15;
if (scanNum + scansToSum < numScans && testSumming)
{
// Get the data for scan scanNum through scanNum + 15
#pragma warning disable 618
reader.GetScanDataSumScans(scanNum, scanNum + scansToSum, out var massIntensityPairs, 0, centroid);
#pragma warning restore 618
Console.WriteLine("Summed spectrum, scans " + scanNum + " through " + (scanNum + scansToSum));
for (var i = 0; i <= massIntensityPairs.GetLength(1) - 1; i += 50)
{
Console.WriteLine(" " + massIntensityPairs[0, i].ToString("0.000") + " mz " +
massIntensityPairs[1, i].ToString("0"));
}
Console.WriteLine();
}
if (!scanInfo.IsFTMS)
{
continue;
}
var dataCount = reader.GetScanLabelData(scanNum, out var ftLabelData);
Console.WriteLine();
Console.WriteLine("{0,12}{1,12}{2,12}{3,12}{4,12}{5,12}", "Mass", "Intensity", "Resolution", "Baseline", "Noise", "Charge");
for (var i = 0; i <= dataCount - 1; i += 50)
{
Console.WriteLine("{0,12:F3}{1,12:0}{2,12:0}{3,12:F1}{4,12:0}{5,12:0}",
ftLabelData[i].Mass,
ftLabelData[i].Intensity,
ftLabelData[i].Resolution,
ftLabelData[i].Baseline,
ftLabelData[i].Noise,
ftLabelData[i].Charge);
}
dataCount = reader.GetScanPrecisionData(scanNum, out var ftPrecisionData);
Console.WriteLine();
Console.WriteLine("{0,12}{1,12}{2,12}{3,12}{4,12}", "Mass", "Intensity", "AccuracyMMU", "AccuracyPPM", "Resolution");
for (var i = 0; i <= dataCount - 1; i += 50)
{
Console.WriteLine("{0,12:F3}{1,12:0}{2,12:F3}{3,12:F3}{4,12:0}",
ftPrecisionData[i].Mass,
ftPrecisionData[i].Intensity, ftPrecisionData[i].AccuracyMMU,
ftPrecisionData[i].AccuracyPPM,
ftPrecisionData[i].Resolution);
}
}
if (mOnlyLoadMSLevelInfo)
{
Console.WriteLine();
Console.WriteLine("{0,-10} {1}", "MSLevel", "Scans");
foreach (var item in msLevelStats)
{
Console.WriteLine("{0, -10} {1}", item.Key, item.Value);
}
}
}
}
catch (Exception ex)
{
ShowError("Error in TestReader: " + ex.Message, ex);
}
}
public void AddMs2ScanParameters(XRawFileIO reader)
{
// grab precursorMz from raw file
var success = reader.GetScanInfo(ScanNumber, out var scanInfo);
if (!success)
{
ConsoleMsgUtils.ShowWarning("Scan {0} not found by AddMs2ScanParameters", ScanNumber);
return;
}
var precursorMzValue = Math.Round(scanInfo.ParentIonMZ, 8);
var parentScanInfo = GetParentScan(reader, scanInfo);
var parentScanDataCount = reader.GetScanData(parentScanInfo.ScanNumber, out var mzList, out var intensityList);
double precursorIntensity = 0;
if (parentScanDataCount > 0)
{
const double PRECURSOR_TOLERANCE = 0.1;
var closestDifference = double.MaxValue;
for (var i = 0; i < mzList.Length; i++)
{
var mzDifference = Math.Abs(mzList[i] - precursorMzValue);
if (mzDifference > closestDifference)
{
continue;
}
closestDifference = mzDifference;
precursorIntensity = Math.Round(intensityList[i], 2);
}
if (closestDifference > PRECURSOR_TOLERANCE)
{
// couldn't find precursor mass in MS1. That's fine i guess.
precursorIntensity = 0;
}
}
var filterLineParts = FilterLine.Split(' ');
string activationType;
switch (scanInfo.ActivationType)
{
case ActivationTypeConstants.CID:
case ActivationTypeConstants.MPD:
case ActivationTypeConstants.ECD:
case ActivationTypeConstants.PQD:
case ActivationTypeConstants.ETD:
case ActivationTypeConstants.HCD:
case ActivationTypeConstants.SA:
case ActivationTypeConstants.PTR:
case ActivationTypeConstants.NETD:
case ActivationTypeConstants.NPTR:
case ActivationTypeConstants.UVPD:
activationType = scanInfo.ActivationType.ToString();
break;
default:
// Includes ActivationTypeConstants.AnyType
activationType = "CID";
break;
}
foreach (var param in filterLineParts)
{
if (param.Contains("@"))
{
// HCD25.00 -> [HCD, 25.00]
// RegEx alpha from numeric
var activationArray = Regex.Matches(param.Split('@')[1].Trim(), @"\D+|\d+").Cast <Match>().Select(m => m.Value).ToArray();
CollisionEnergy = Convert.ToInt32(double.Parse(activationArray[1]));
}
}
PrecursorMz = new PrecursorMz(precursorIntensity, activationType, precursorMzValue);
}
private bool LoadSpectraForThermoRawFile(
XRawFileIO xcaliburAccessor,
clsSpectraCache spectraCache,
clsScanInfo scanInfo,
clsBaselineNoiseOptions noiseThresholdOptions,
bool discardLowIntensityData,
bool compressSpectraData,
double msDataResolution,
bool keepRawSpectrum)
{
var lastKnownLocation = "Start";
try
{
// Load the ions for this scan
lastKnownLocation = "xcaliburAccessor.GetScanData for scan " + scanInfo.ScanNumber;
// Retrieve the m/z and intensity values for the given scan
// We retrieve the profile-mode data, since that's required for determining spectrum noise
scanInfo.IonCountRaw = xcaliburAccessor.GetScanData(scanInfo.ScanNumber, out var mzList, out var intensityList);
if (scanInfo.IonCountRaw > 0)
{
var ionCountVerified = VerifyDataSorted(scanInfo.ScanNumber, scanInfo.IonCountRaw, mzList, intensityList);
if (ionCountVerified != scanInfo.IonCountRaw)
{
scanInfo.IonCountRaw = ionCountVerified;
}
}
scanInfo.IonCount = scanInfo.IonCountRaw;
lastKnownLocation = "Instantiate new clsMSSpectrum";
var msSpectrum = new clsMSSpectrum(scanInfo.ScanNumber, mzList, intensityList, scanInfo.IonCountRaw);
lastKnownLocation = "Manually determine the base peak m/z and base peak intensity";
// ReSharper disable once CommentTypo
// Regarding BPI, comparison of data read via the ThermoRawFileReader vs.
// that read from the .mzML file for dataset QC_Shew_18_02-run1_02Mar19_Arwen_18-11-02
// showed that 25% of the spectra had incorrect BPI values
double totalIonIntensity = 0;
double basePeakIntensity = 0;
double basePeakMz = 0;
for (var ionIndex = 0; ionIndex < scanInfo.IonCountRaw; ionIndex++)
{
totalIonIntensity += intensityList[ionIndex];
if (intensityList[ionIndex] > basePeakIntensity)
{
basePeakIntensity = intensityList[ionIndex];
basePeakMz = mzList[ionIndex];
}
}
if (Math.Abs(scanInfo.BasePeakIonMZ - basePeakMz) > 0.1)
{
mBpiUpdateCount += 1;
if (mBpiUpdateCount < 10)
{
ConsoleMsgUtils.ShowDebug("Updating BPI in scan {0} from {1:F3} m/z to {2:F3} m/z, and BPI Intensity from {3:F0} to {4:F0}",
scanInfo.ScanNumber, scanInfo.BasePeakIonMZ, basePeakMz, scanInfo.BasePeakIonIntensity, basePeakIntensity);
}
scanInfo.BasePeakIonMZ = basePeakMz;
scanInfo.BasePeakIonIntensity = basePeakIntensity;
}
// Determine the minimum positive intensity in this scan
lastKnownLocation = "Call mMASICPeakFinder.FindMinimumPositiveValue";
scanInfo.MinimumPositiveIntensity = mPeakFinder.FindMinimumPositiveValue(msSpectrum.IonsIntensity, 0);
if (msSpectrum.IonCount > 0)
{
if (scanInfo.TotalIonIntensity < float.Epsilon)
{
scanInfo.TotalIonIntensity = totalIonIntensity;
}
}
else
{
scanInfo.TotalIonIntensity = 0;
}
bool discardLowIntensityDataWork;
bool compressSpectraDataWork;
if (scanInfo.MRMScanType == MRMScanTypeConstants.NotMRM)
{
discardLowIntensityDataWork = discardLowIntensityData;
compressSpectraDataWork = compressSpectraData;
}
else
{
discardLowIntensityDataWork = false;
compressSpectraDataWork = false;
}
lastKnownLocation = "Call ProcessAndStoreSpectrum";
mScanTracking.ProcessAndStoreSpectrum(
scanInfo, this,
spectraCache, msSpectrum,
noiseThresholdOptions,
discardLowIntensityDataWork,
compressSpectraDataWork,
msDataResolution,
keepRawSpectrum);
}
catch (Exception ex)
{
ReportError("Error in LoadSpectraForThermoRawFile (LastKnownLocation: " + lastKnownLocation + ")", ex, clsMASIC.eMasicErrorCodes.InputFileDataReadError);
return(false);
}
return(true);
}
private double ComputeInterference(
XRawFileIO xcaliburAccessor,
ThermoRawFileReader.clsScanInfo scanInfo,
int precursorScanNumber)
{
if (precursorScanNumber != mCachedPrecursorScan)
{
xcaliburAccessor.GetScanData(precursorScanNumber, out var centroidedIonsMz, out var centroidedIonsIntensity, 0, true);
UpdateCachedPrecursorScan(precursorScanNumber, centroidedIonsMz, centroidedIonsIntensity);
}
var chargeState = 0;
scanInfo.TryGetScanEvent(SCAN_EVENT_CHARGE_STATE, out var chargeStateText, true);
if (!string.IsNullOrWhiteSpace(chargeStateText))
{
if (!int.TryParse(chargeStateText, out chargeState))
{
chargeState = 0;
}
}
if (!scanInfo.TryGetScanEvent(SCAN_EVENT_MS2_ISOLATION_WIDTH, out var isolationWidthText, true))
{
if (scanInfo.MRMScanType == MRMScanTypeConstants.SRM)
{
// SRM data files don't have the MS2 Isolation Width event
return(0);
}
WarnIsolationWidthNotFound(
scanInfo.ScanNumber,
"Could not determine the MS2 isolation width (" + SCAN_EVENT_MS2_ISOLATION_WIDTH + ")");
return(0);
}
if (!double.TryParse(isolationWidthText, out var isolationWidth))
{
ReportWarning("MS2 isolation width (" + SCAN_EVENT_MS2_ISOLATION_WIDTH + ") was non-numeric (" + isolationWidthText + "); " +
"cannot compute interference for scan " + scanInfo.ScanNumber);
return(0);
}
double parentIonMz;
if (Math.Abs(scanInfo.ParentIonMZ) > 0)
{
parentIonMz = scanInfo.ParentIonMZ;
}
else
{
// ThermoRawFileReader could not determine the parent ion m/z value (this is highly unlikely)
// Use scan event "Monoisotopic M/Z" instead
if (!scanInfo.TryGetScanEvent(SCAN_EVENT_MONOISOTOPIC_MZ, out var monoMzText, true))
{
ReportWarning("Could not determine the parent ion m/z value (" + SCAN_EVENT_MONOISOTOPIC_MZ + "); " +
"cannot compute interference for scan " + scanInfo.ScanNumber);
return(0);
}
if (!double.TryParse(monoMzText, out var mz))
{
OnWarningEvent(string.Format("Skipping scan {0} since scan event {1} was not a number: {2}",
scanInfo.ScanNumber, SCAN_EVENT_MONOISOTOPIC_MZ, monoMzText));
return(0);
}
parentIonMz = mz;
}
if (Math.Abs(parentIonMz) < float.Epsilon)
{
ReportWarning("Parent ion m/z is 0; cannot compute interference for scan " + scanInfo.ScanNumber);
return(0);
}
var precursorInterference = ComputePrecursorInterference(
scanInfo.ScanNumber,
precursorScanNumber, parentIonMz, isolationWidth, chargeState);
return(precursorInterference);
}
public List <CompoundData> ReadSpectraData(List <CompoundData> combinedTransitions)
{
var scanTimes = new Dictionary <int, double>();
using (var rawReader = new XRawFileIO(DatasetPath))
{
var numScans = rawReader.GetNumScans();
if (numScans <= 0)
{
// dataset has no MS data. Return.
return(null);
}
for (var i = 1; i <= numScans; i++)
{
var err = rawReader.GetRetentionTime(i, out double time);
scanTimes.Add(i, time);
// Filter string parsing for speed: "+ c NSI SRM ms2 [precursor/parent m/z] [[list of product m/z ranges]]
}
// Map transition start/stop times to scans
var scansAndTargets = new Dictionary <int, List <CompoundData> >();
foreach (var scan in scanTimes)
{
var matches = combinedTransitions
.Where(x => x.StartTimeMinutes <= scan.Value && scan.Value <= x.StopTimeMinutes).ToList();
if (matches.Count > 0)
{
scansAndTargets.Add(scan.Key, matches);
}
}
rawReader.ScanInfoCacheMaxSize = 2;
// read spectra data for each transition from the file, in an optimized fashion
foreach (var scan in scansAndTargets)
{
rawReader.GetScanInfo(scan.Key, out clsScanInfo scanInfo);
var preMz = scanInfo.ParentIonMZ;
rawReader.GetScanData(scan.Key, out var mzs, out var intensities);
foreach (var compound in scan.Value.Where(x => Math.Abs(x.PrecursorMz - preMz) < 0.01))
{
foreach (var massRange in scanInfo.MRMInfo.MRMMassList)
{
var match = compound.Transitions.FirstOrDefault(x => massRange.StartMass <= x.ProductMz && x.ProductMz <= massRange.EndMass);
if (match == null)
{
continue;
}
for (var j = 0; j < mzs.Length; j++)
{
var mz = mzs[j];
var intensity = intensities[j];
if (massRange.StartMass <= mz && mz <= massRange.EndMass)
{
match.IntensitySum += intensity;
match.Intensities.Add(new TransitionData.DataPoint(scan.Key, scanTimes[scan.Key], intensity));
if (intensity > match.MaxIntensity)
{
match.MaxIntensity = intensity;
match.MaxIntensityTime = scanTimes[scan.Key];
}
}
}
}
}
}
}
return(combinedTransitions.ToList());
}