void BFReference::CalcSignalReference2(const std::string &datFile, const std::string &bgFile, Mask &mask, int traceFrame) { Image bfImg; Image bfBkgImg; bfImg.SetImgLoadImmediate (false); bfBkgImg.SetImgLoadImmediate (false); bool loaded = bfImg.LoadRaw(datFile.c_str()); bool bgLoaded = bfBkgImg.LoadRaw(bgFile.c_str()); if (!loaded) { ION_ABORT("*Error* - No beadfind file found, did beadfind run? are files transferred? (" + datFile + ")"); } if (!bgLoaded) { ION_ABORT("*Error* - No beadfind background file found, did beadfind run? are files transferred? (" + bgFile + ")"); } const RawImage *raw = bfImg.GetImage(); assert(raw->cols == GetNumCol()); assert(raw->rows == GetNumRow()); assert(raw->cols == mask.W()); assert(raw->rows == mask.H()); int StartFrame = bfImg.GetFrame(-663); //5 int EndFrame = bfImg.GetFrame(350); //20 int NNinnerx = 1, NNinnery = 1, NNouterx = 12, NNoutery = 8; cout << "DC start frame: " << StartFrame << " end frame: " << EndFrame << endl; bfImg.FilterForPinned(&mask, MaskEmpty, false); bfImg.XTChannelCorrect(); // bfImg.XTChannelCorrect(&mask); Traces trace; trace.Init(&bfImg, &mask, FRAMEZERO, FRAMELAST, FIRSTDCFRAME,LASTDCFRAME); bfImg.Normalize(StartFrame, EndFrame); if (mDoRegionalBgSub) { trace.SetMeshDist(0); } trace.CalcT0(true); if (mDoRegionalBgSub) { GridMesh<float> grid; grid.Init(raw->rows, raw->cols, mRegionYSize, mRegionXSize); int numBin = grid.GetNumBin(); int rowStart = -1, rowEnd = -1, colStart = -1, colEnd = -1; for (int binIx = 0; binIx < numBin; binIx++) { cout << "BG Subtract Region: " << binIx << endl; grid.GetBinCoords(binIx, rowStart, rowEnd, colStart, colEnd); Region reg; reg.row = rowStart; reg.h = rowEnd - rowStart; reg.col = colStart; reg.w = colEnd - colStart; bfImg.BackgroundCorrectRegion(&mask, reg, MaskAll, MaskEmpty, NNinnerx, NNinnery, NNouterx, NNoutery, NULL); } } else { bfImg.BackgroundCorrect(&mask, MaskEmpty, MaskEmpty, NNinnerx, NNinnery, NNouterx, NNoutery, NULL); } int length = GetNumRow() * GetNumCol(); mBfMetric.resize(length, std::numeric_limits<double>::signaling_NaN()); for (int wIx = 0; wIx < length; wIx++) { if (mask[wIx] & MaskExclude || mask[wIx] & MaskPinned) continue; int t0 = (int)trace.GetT0(wIx); mBfMetric[wIx] = 0; float zSum = 0; int count = 0; for (int fIx = min(t0-20, 0); fIx < t0-10; fIx++) { zSum += bfImg.At(wIx,fIx); count ++; } for (int fIx = t0+3; fIx < t0+15; fIx++) { mBfMetric[wIx] += (bfImg.At(wIx,fIx) - (zSum / count)); } } bfImg.Close(); for (int i = 0; i < length; i++) { if (mask[i] & MaskExclude || mWells[i] == Exclude) { mWells[i] = Exclude; } else { mask[i] = MaskIgnore; } } cout << "Filling reference. " << endl; FillInReference(mWells, mBfMetric, mGrid, mMinQuantile, mMaxQuantile, mNumEmptiesPerRegion); for (int i = 0; i < length; i++) { if (mWells[i] == Reference) { mask[i] = MaskEmpty; } } }
void BFReference::CalcReference(const std::string &datFile, Mask &mask, std::vector<float> &metric) { Image bfImg; bfImg.SetImgLoadImmediate (false); bool loaded = bfImg.LoadRaw(datFile.c_str()); if (!loaded) { ION_ABORT("*Error* - No beadfind file found, did beadfind run? are files transferred? (" + datFile + ")"); } const RawImage *raw = bfImg.GetImage(); assert(raw->cols == GetNumCol()); assert(raw->rows == GetNumRow()); assert(raw->cols == mask.W()); assert(raw->rows == mask.H()); if (!mDebugFile.empty()) { DebugTraces(mDebugFile, mask, bfImg); } bfImg.FilterForPinned(&mask, MaskEmpty, false); // int StartFrame= bfImg.GetFrame((GetDcStart()*1000/15)-1000); // int EndFrame = bfImg.GetFrame((GetDcEnd()*1000/15)-1000); int StartFrame = bfImg.GetFrame(-663); //5 int EndFrame = bfImg.GetFrame(350); //20 cout << "DC start frame: " << StartFrame << " end frame: " << EndFrame << endl; bfImg.XTChannelCorrect(); FilterForOutliers(bfImg, mask, mIqrOutlierMult, mRegionYSize, mRegionXSize); bfImg.Normalize(StartFrame, EndFrame); // bfImg.XTChannelCorrect(&mask); int NNinnerx = 1, NNinnery = 1, NNouterx = 12, NNoutery = 8; if (mDoRegionalBgSub) { GridMesh<float> grid; grid.Init(raw->rows, raw->cols, mRegionYSize, mRegionXSize); int numBin = grid.GetNumBin(); int rowStart = -1, rowEnd = -1, colStart = -1, colEnd = -1; for (int binIx = 0; binIx < numBin; binIx++) { grid.GetBinCoords(binIx, rowStart, rowEnd, colStart, colEnd); Region reg; reg.row = rowStart; reg.h = rowEnd - rowStart; reg.col = colStart; reg.w = colEnd - colStart; bfImg.BackgroundCorrectRegion(&mask, reg, MaskAll, MaskEmpty, NNinnerx, NNinnery, NNouterx, NNoutery, NULL); } } else { bfImg.BackgroundCorrect(&mask, MaskEmpty, MaskEmpty, NNinnerx, NNinnery, NNouterx, NNoutery, NULL); } Region region; region.col = 0; region.row = 0; region.w = GetNumCol(); //mGrid.GetColStep(); region.h = GetNumRow(); // mGrid.GetRowStep(); int startFrame = bfImg.GetFrame(12); // frame 15 on uncompressed 314 // int endFrame = bfImg.GetFrame(raw->timestamps[bfImg.Ge]5300); // frame 77 or so int endFrame = bfImg.GetFrame(5000); // frame 77 or so bfImg.CalcBeadfindMetric_1(&mask, region, "pre", startFrame, endFrame); const double *results = bfImg.GetResults(); int length = GetNumRow() * GetNumCol(); metric.resize(length); copy(&results[0], &results[0] + (length), metric.begin()); bfImg.Close(); }