CoralspeQ | all-in-one protocol


Derive Phi2 and color from a series of reflectance spectra. Original trace is added back.
//============================================
// Macro for data evaluation on PhotosynQ.org
// created: 4/17/2016
//============================================

//Define the output object here
var output = {};

// Number of pixel per spectrum 
var pixels = 256;

// Number of spectra collected
var specNum = 16;

// Background Spectrum
var bgspec = 12;

// Integration times for each spectrum
// The integration times are currently taken from the protocol
var intTimes = [256,128,64,256,128,64,32,16,8,16,256,16,256,25,50,100];

// Pixel reflecting 730 nm
var pix = 80;
var A0 = 3.2e2;
var B1 = 2.4;
var B2 = -9e-4;
var B3 = -4e-6;
var B4 = -3e-9;
var B5 = 2e-11;

var fallbackBlack = [8608,8617,8623,8627,8639,8610,8639,8647,8612,8592,8593,8595,8607,8621,8606,8495,8448,8601,8629,8628,8625,8635,8638,8670,8659,8668,8688,8710,8779,8868,9053,9427,10127,11329,13053,14985,17557,19097,19563,18849,17281,15645,13600,12468,11328,10678,10112,9759,9466,9349,9301,9259,9302,9408,9454,9645,9929,10024,10085,10034,9900,9736,9600,9494,9393,9333,9272,9230,9164,9172,9172,9193,9222,9182,9149,9373,9412,9463,9504,9552,9567,9605,9613,9626,9654,9649,9673,9668,9674,9712,9753,9774,9786,9805,9985,9883,9903,9929,9955,9957,10010,10044,10098,10138,10194,10213,10247,10291,10345,10362,10418,10472,10422,10702,10593,10646,10680,10736,10748,10791,10795,10825,10852,10876,10882,10928,10947,10965,10970,10962,10968,10983,11230,10962,10951,11118,10829,10872,10822,10980,10823,10808,10773,10749,10733,10674,10614,10571,10513,10479,10430,10297,10249,10251,10262,10179,10136,10068,10028,9983,9925,9863,9836,9782,9716,9674,9616,9582,9528,9504,9460,9971,9370,9366,9331,9338,9268,9228,9185,9122,9120,9100,9069,9054,9034,9021,9005,8956,8958,8920,8800,8694,8879,8852,8854,8836,8814,8812,8798,8796,8788,8760,8766,8725,8741,8706,8717,8706,8687,8676,8333,8706,8675,8668,8676,8681,8680,8683,8664,8638,8650,8642,8646,8644,8643,8652,8639,8626,8777,8368,8643,8543,8654,8641,8623,8603,8593,8604,8590,8606,8618,8586,8611,8608,8613,8592,8623,8596,8490,8553,8616,8599,8575,8597,8612,8583];
var fallbackWhite = [8691,8698,8708,8686,8694,8688,8692,8686,8679,8662,8684,8673,8676,8714,8690,8573,8466,8684,8681,8705,8716,8711,8731,8730,8756,8753,8781,8825,8867,9005,9215,9645,10561,12433,15684,20111,25193,28943,30495,29709,27092,23935,20344,17927,15651,14162,12942,12132,11642,11418,11400,11535,11927,12612,13399,14490,15867,16858,17321,17180,16485,15577,14667,13896,13306,12850,12479,12099,11934,11789,11779,11897,12107,12283,12412,12984,13342,13648,13916,14185,14393,14569,14660,14776,14845,14908,14989,15045,15139,15274,15422,15581,15782,15872,16211,16264,16382,16547,16675,16829,17032,17278,17574,17844,18072,18322,18566,18805,19051,19322,19643,19951,20188,20728,20895,21189,21451,21649,21817,21963,22181,22391,22654,22895,23173,23421,23641,23818,23939,23971,24000,23998,24119,23967,23941,24043,23799,23836,23801,23892,23722,23611,23424,23210,22969,22685,22396,22109,21854,21618,21407,21052,20903,20679,20493,20234,19921,19587,19239,18878,18525,18210,17868,17540,17195,16867,16561,16199,15872,15579,15316,15837,14803,14592,14351,14111,13871,13587,13338,13090,12863,12628,12494,12321,12132,12016,11843,11660,11506,11319,11049,10761,10846,10720,10585,10466,10372,10253,10179,10069,9970,9884,9772,9707,9619,9577,9523,9511,9452,9397,9095,9361,9327,9324,9295,9244,9238,9226,9191,9169,9138,9126,9087,9075,9040,9034,9009,8986,9187,8659,8929,8854,8982,8915,8887,8842,8853,8844,8818,8781,8818,8799,8796,8765,8772,8780,8766,8749,8660,8577,8777,8734,8714,8716,8700,8707];
var fallbackInt = [256,128,64,256,128,64,32,16,8,16,256,16,256,100,200,400];
var fallback = false;


if(json.get_userdef1 === undefined){
  json.get_userdef1 = [100,100];
  intTimes = fallbackInt;
  fallback = true;
}
if(json.get_userdef51 === undefined)
  json.get_userdef51 = fallbackBlack;
if(json.get_userdef52 === undefined)
  json.get_userdef52 = fallbackWhite;

//output.pixel = A0 + B1*pix + B2*Math.pow(pix,2) + B3*Math.pow(pix,3) + B4*Math.pow(pix,4) + B5*Math.pow(pix,5);

var GFP_Ref = [-0.000453666,-0.002343933,-0.004782138,-0.007079998,-0.008823949,-0.009957256,-0.010321658,-0.011075738,-0.010859356,-0.010683285,-0.009910008,-0.010220661,-0.009372732,-0.008397897,-0.007867981,-0.007175326,-0.006036259,-0.005191317,-0.004867437,-0.003361087,-0.002411634,-0.001341245,-4.24249E-05,0.000851787,0.002692573,0.003403144,0.005030428,0.006865242,0.009134524,0.01122763,0.014307621,0.01665454,0.020770686,0.026130516,0.032130851,0.040991808,0.055512799,0.076065582,0.109592695,0.162608175,0.248582033,0.475371402,1,1.02126794,1.033747974,1.046319082,1.05621918,1.068676819,1.079580226,1.090609046,1.101606513,1.113623711,1.124568923,1.135131921,1.141441311,1.145021463,1.145296072,1.142220349,1.132420072,1.119909966,1.103476208,1.088299572,1.071655298,1.0598708,1.04848643,1.037299138,1.029009795,1.020638336,1.01378229,1.006675418,1.000032874,0.994866925,0.990443006,0.986956703,0.982146092,0.971308167,0.927795537,0.592327663,0.47032557,0.378274916,0.304697377,0.247087669,0.201262353,0.163933805,0.135049768,0.112316966,0.094920211,0.079144875,0.066233147,0.054984642,0.046186179,0.038437309,0.031897783,0.026427258,0.020529731,0.015305554,0.010986147,0.00666226,0.00377615,0.00066758,-0.000695651,-0.001124253,-0.001336367,-0.001899341,-0.002008437,-0.002098124,-0.002423707,-0.001889312,-0.002316421,-0.001941779,-0.001084893,-0.000958092,-0.001444921,-0.000485016,0.000313643,-0.000739041,-0.00084963,-0.000746717,-0.001149938,-0.001390419,-0.001327818,-0.000382843,-0.001698298,-0.001078801,-0.000965437,-0.001595597,-0.001358313,-0.001156861,-0.000815065,-0.001079435,-0.000988467,-0.000778057,-0.000924478,-0.000772296,-0.000539491,-0.000518694,-0.000541195,9.17388E-05,-0.000245789,-0.000540019,0.000261626,3.75675E-05,-9.39238E-05,0.000168742,0.000283598,0.00029693,-0.000231704,-9.86523E-05,3.46109E-05,-0.000238227,-0.000418498,-0.000779423,-0.000380403,-0.000760738,0.000501247,-0.000467337,-0.000752119,-0.000826386,-0.000619965,-0.001119741,-0.001249249,-0.001217511,-0.001379866,-0.001363058,0.000242321,-0.001677316,-0.001608252,-0.001661616,-0.00211063,-0.002140106,-0.002102396,-0.002542452,-0.003043721,-0.002735775,-0.002677163,-0.002891773,-0.002919756,-0.00316124,-0.003035442,-0.002857388,-0.003164565,-0.002986512,-0.003050327,-0.002846892,-0.002522522,-0.002154855,-0.002560572,-0.002858791,-0.002883787,-0.002574348,-0.002593373,-0.002884127,-0.002416427,-0.002607149,-0.002874015,-0.003243899,-0.002983729,-0.003144591,-0.002797826,-0.002507796,-0.002941879,-0.003069894,-0.002660423,-0.002798889,-0.002923918,-0.002577153,-0.002490173,-0.002664472,-0.002378921,-0.002744326,-0.002597626,-0.001985104,-0.002449048,-0.00231877,-0.001961555,-0.001958184,-0.00152781,-0.001976824,-0.001922691,-0.002247784,-0.001938345,-0.002370936,-0.002100315,-0.001487793,-0.002275722,-0.002002115,-0.001753889,-0.001714686,-0.001878534,-0.001793047,-0.002006164,-0.001780334,-0.001121528,-0.001394366,-0.001634357,-0.001233844,-0.001693309,-0.001485395,-0.00101023,-0.00116512,-0.001255809,-0.001189732,0.001078547,-0.001223302,-0.001128858,-0.00130017,-0.000698099,-0.001341205,-0.001022808,-0.000668578,-6.78645E-06,-0.000409517,-0.000232956,-0.000228092,-0.00036208,-5.26402E-05]


var nm = {};
var nms = [];
var pixel = {};
for(i=0;i<256;i++){
   var pix = i+1;
   var wl = A0 + B1*pix + B2*Math.pow(pix,2) + B3*Math.pow(pix,3) + B4*Math.pow(pix,4) + B5*Math.pow(pix,5);
   nm[MathROUND(wl,0)] = pix;
   nms.push(MathROUND(wl,0));
   pixel[pix] = [MathROUND(wl,0)];
}


// Now we fill up the array to make sure the is no undefiend when we enter a wavelength
var min = MathMIN(nms);
var max = MathMAX(nms);
var nms = {};
var tmp = 0;
for(var i = min;i<max;i++){
	if(nm[i] !== undefined){
      nms[i] = nm[i];
		tmp	= i;
    }
    else{
		nms[i] = nm[tmp];
    }
}
var nm = nms;


// Split the rawtrace up into separate spectra
// The number of spectra is set in specNum.
// The traces gets devided by the number of pixels for a spectrum
// defined by pixels
var spectra = [];
for (i=0,j=json.data_raw.length; i<j; i+=pixels) {
    spectra.push(json.data_raw.slice(i,i+pixels));   
}


// Normalize spectra to integration times
for(var i=0;i<bgspec+1;i++){
  for(var j=0;j<pixels;j++){
	spectra[i][j] = spectra[i][j] / intTimes[i];
  }
}

// Substract background spectrum from all other spectra
for(var i=0;i<bgspec;i++){
  for(var j=0;j<pixels;j++){
	spectra[i][j] -= spectra[bgspec][j]; 
  }
}

// Clone spectrum #12 here for the Chloropyll fluorecence ratio
var f_ratio_spectrum = spectra[11].slice(0);
var GFP_ratio_spectrum = spectra[11].slice(0);

var ms = [];
//Substract drift
for(var i=0;i<bgspec;i++){
  var len = spectra[i].length;
  var m = (spectra[i][nm[650]] - spectra[i][0]) / (nm[650] - 0);
  var b = spectra[i][nm[650]] - (m * nm[650]);
  for(var j=0;j<len;j++){
	spectra[i][j] -= (m * j) + b;
  }
}


//Substract drift
for(var i=0;i<bgspec;i++){
  var len = spectra[i].length;
  var m = (spectra[i][len-1] - spectra[i][nm[650]]) / (len - nm[650]);
  var b = spectra[i][nm[650]] - (m * nm[650]);
  for(var j=nm[650];j<len;j++){
	spectra[i][j] -= (m * j) + b;
  }
}


var m = spectra[11].slice(nm[420],nm[525]);
var min = MathMIN( m );
var max = MathMAX( m );

var lower = 420;
for(i=420;i<525;i++){
	var IntInPercent = ((spectra[11][nm[i]] - min) / max)  * 100;
	if(IntInPercent < 15)
		lower = i;
    else
		break;
}


var higher = 525;
for(i=525;i>420;i--){
	var IntInPercent = ((spectra[11][nm[i]] - min) / max)  * 100;
	if(IntInPercent < 15)
		higher = i;
    else
		break;
}

//Normalize to 410-490 better 510-530
for(var i=0;i<bgspec;i++){
  var n = spectra[i].slice(nm[425],nm[430]);
  var m = spectra[i].slice(nm[495],nm[505]);
  var o = spectra[i].slice(nm[425],nm[505]);
  var norm = MathMEAN(n) + MathMEAN(m);
  var norm = spectra[i][nm[420]];
  for(var j=0;j<pixels;j++){
	spectra[i][j] = spectra[i][j] / norm;
  }
}

var nmMin = 700;
var nmMax = 780;

var x = [];
var y = [];

for(var i=3;i<bgspec-1;i++){
  x.push( json.tcs_dac_values[i] );
  y.push( MathSUM(spectra[i].slice(nm[nmMin],nm[nmMax])) );
}
var reg = MathLINREG(x.slice(1,4), y.slice(1,4));

// Fm' is derived from the last spectrum 
output.Fmp = MathSUM(spectra[11].slice(nm[nmMin],nm[nmMax]));

// Fs extrapolated and r^2
output.Fs = reg.b;
output.r2 = MathROUND(reg.r2, 3);

for (i=0; i<pixels; i++){
	var tmp = [];
    tmp.push(pixel[i+1]);
	
	tmp.push( (spectra[11][i] - spectra[0][i]) / spectra[11][i]);
	//output[pixel[i+1]] = tmp.join('=');
}

output.Fs = MathROUND(output.Fs,3);
output.Fmp = MathROUND(output.Fmp,3);
output['ΦII'] = MathROUND( (output.Fmp - output.Fs)/output.Fmp, 3);

if(output.Fs < 0 || output.Fmp < 0 || output['ΦII'] < 0)
  output['ΦII'] = 'NaN';


// In case the number of spectra in specNum and the number after the
// trace got split didn't match up, a warning is triggered
if(spectra.length != specNum)
	danger('Insufficent number of spectra collected!',output);



/*
///////////////////////////////////////////////////////////////
/
/				Trying to extract the GFP signal
/
///////////////////////////////////////////////////////////////
*/

var GFP_REF_R = GFP_Ref.slice(100,120);
var GFP_REF_G = GFP_Ref.slice(80,100);
var GFP_REF_B = GFP_Ref.slice(80,88);

var GFP_R = spectra[11].slice(100,120);
var GFP_G = spectra[11].slice(80,100);
var GFP_B = spectra[11].slice(80,88);

var GFP_Red = MathMEAN(GFP_R) - MathMEAN(GFP_REF_R);
var GFP_Green = MathMEAN(GFP_G) - MathMEAN(GFP_REF_G);
var GFP_Blue = MathMEAN(GFP_B) - MathMEAN(GFP_REF_B);

var gfp_gamma = 1.5;

var GFP_Red = Math.pow( GFP_Red,(1/gfp_gamma) ) * 255     || 0;
var GFP_Green = Math.pow( GFP_Green,(1/gfp_gamma) ) * 255 || 0;
var GFP_Blue = Math.pow( GFP_Blue,(1/gfp_gamma) ) * 255   || 0;

if(GFP_Red > 255 || GFP_Green > 255 || GFP_Blue > 255){

  var F_max = MathMAX([GFP_Blue,GFP_Green,GFP_Red]);
  
  GFP_Red *= (255/F_max) || 0;
  GFP_Green *= (255/F_max) || 0;
  GFP_Blue *= (255/F_max) || 0; 
}

GFP_Red = MathROUND(GFP_Red,0);
GFP_Green = MathROUND(GFP_Green,0);
GFP_Blue = MathROUND(GFP_Blue,0);


GFP_Red = (GFP_Red < 0 || isNaN(GFP_Red)) ? 0 : GFP_Red;
GFP_Green = (GFP_Green < 0 || isNaN(GFP_Green)) ? 0 : GFP_Green;
GFP_Blue = 0;//(GFP_Blue < 0 || isNaN(GFP_Blue)) ? 0 : GFP_Blue;


output.GFP = 'rgb('+GFP_Red+','+GFP_Green+','+GFP_Blue+')';

output.GFP_Spectrum = spectra[11].slice(80,150);

//output.GFP_Red = GFP_Red;
//output.GFP_Green = GFP_Green;
//output.GFP_Blue = GFP_Blue;

/*
///////////////////////////////////////////////////////////////
/
/			Spectral shifts in the chlorophyll
/
///////////////////////////////////////////////////////////////
*/

f_ratio_spectrum = spectra[11].slice(0);

var f_ratio_a = f_ratio_spectrum.slice(160,190);
var f_ratio_b = f_ratio_spectrum.slice(200,230);

var f_ratio_a_max = MathMEAN(f_ratio_a.sort().slice(27,30));
var f_ratio_b_max = MathMEAN(f_ratio_b.sort().slice(27,30));

output['A/B ratio'] = MathROUND( (f_ratio_a_max/f_ratio_b_max), 3);

var f_red = f_ratio_spectrum.slice(160,190);
var f_green = f_ratio_spectrum.slice(200,230);
var f_blue = f_ratio_spectrum.slice(160,190);

var fredcolor = spectra[11].slice(157,170);
var fgreencolor = spectra[11].slice(170,190);
var fbluecolor = spectra[11].slice(190,230);

for(i=0;i<pixels;i++){
  if(fredcolor[i] !== undefined)
      fredcolor[i] = fredcolor[i] / spectra[11][nm[690]];
  if(fgreencolor[i] !== undefined)
      fgreencolor[i] = fgreencolor[i] / spectra[11][nm[690]];
  if(fbluecolor[i] !== undefined)
      fbluecolor[i] = fbluecolor[i] / spectra[11][nm[690]];
}

var f_gamma = 1;

var F_Red = MathMEAN(fredcolor);
var F_Green = MathMEAN(fgreencolor);
var F_Blue = MathMEAN(fbluecolor);

F_Red = Math.pow(F_Red,(1/f_gamma) ) * 255      || 0;
F_Green = Math.pow(F_Green,(1/f_gamma) ) * 255  || 0;
F_Blue = Math.pow(F_Blue,(1/f_gamma) ) * 255    || 0;

if(F_Red > 255 || F_Green > 255 || F_Blue > 255){

  var F_max = MathMAX([F_Blue,F_Green,F_Red]);
  
  F_Red *= (255/F_max);
  F_Green *= (255/F_max);
  F_Blue *= (255/F_max);
}

F_Red = MathROUND(F_Red,0);
F_Green = MathROUND(F_Green,0);
F_Blue = MathROUND(F_Blue,0);
  

F_Red = (F_Red < 0 || isNaN(F_Red)) ? 0 : F_Red;
F_Green = (F_Green < 0 || isNaN(F_Green)) ? 0 : F_Green;
F_Blue = (F_Blue < 0 || isNaN(F_Blue)) ? 0 : F_Blue;


output.Fluorescence = 'rgb('+F_Red+','+F_Green+','+F_Blue+')';

output.F_Red = F_Red;
output.F_Green = F_Green;
output.F_Blue = F_Blue;

var test = f_ratio_spectrum.slice(150,230);
var test_n = test[14];
for(i=0;i<test.length;i++){
	test[i] = test[i] / test_n;
}

output.F_Spectrum = test;


/*
///////////////////////////////////////////////////////////////
/
/	Estimation of the color from a reflectance spectrum
/
///////////////////////////////////////////////////////////////
*/


// Spectrum used to determine RGB color
var SpectrumForRGB = 15;

// Pick a non saturated spectrum
if(MathMAX(spectra[SpectrumForRGB]) > 40000)
	SpectrumForRGB = 14;

if(MathMAX(spectra[SpectrumForRGB]) > 40000)
	SpectrumForRGB = 13;

if(fallback)
  SpectrumForRGB = 13;

var Sample = [];
var Black = [];
var White = [];


var SampleInt = intTimes[SpectrumForRGB];
var BlackInt = json.get_userdef1[0];
var WhiteInt = json.get_userdef1[1];

var SampleRef = [];
var ReferenceRef = [];

for(i=0;i<pixels;i++){
	// intTimes[SpectrumForRGB];
    Sample.push(spectra[SpectrumForRGB][i]);
    // /BlackInt;
    Black.push(json.get_userdef51[i]);
    // /WhiteInt;
    White.push(json.get_userdef52[i]);
}

var sample_off = MathMEAN(Sample.slice(10,20));
var black_off = MathMEAN(Black.slice(10,20));
var white_off = MathMEAN(White.slice(10,20));
    
    
// Let's substract the offset here
for(i=0;i<pixels;i++){
  Sample[i] -= sample_off;
  Black[i] -= black_off;
  White[i] -= white_off;
}

for(i=0;i<pixels;i++){
    SampleRef.push( ((Sample[i] / SampleInt) - (Black[i]  / BlackInt) ) );
	ReferenceRef.push( ((White[i] / WhiteInt) - (Black[i]  / BlackInt) ) );
}

// Calculate reflectance spectra
var ReflectanceSpectrum = [];


for(i=0;i<pixels;i++){
    ReflectanceSpectrum.push(SampleRef[i] / ReferenceRef[i]);
}

//var offset = MathMEAN(ReflectanceSpectrum.slice(32,42)); // 32,42
//var ReflectanceSpectrumOffset = [];


//for(i=0;i<pixels;i++){
    //var value = (ReflectanceSpectrum[i] - offset) < 0 ? 0 : (ReflectanceSpectrum[i] - offset);
    //var value = (ReflectanceSpectrum[i] - offset);
    //ReflectanceSpectrumOffset.push(value);
//}

// Calculate corrected relectance spectrum against 730nm
var ReflectanceSpectrumOffsetNorm = [];

for(i=0;i<pixels;i++){
	ReflectanceSpectrumOffsetNorm.push( (ReflectanceSpectrum[i] / ReflectanceSpectrum[nm[730]]) );
}

// Calculate the RGB color
//var redSpec = ReflectanceSpectrumOffsetNorm.slice(nm[630],nm[730]);
//var greenSpec = ReflectanceSpectrumOffsetNorm.slice(nm[530],nm[580]);
//var blueSpec = ReflectanceSpectrumOffsetNorm.slice(nm[440],nm[475]);

var redSpec = ReflectanceSpectrumOffsetNorm.slice(142,199);
var greenSpec = ReflectanceSpectrumOffsetNorm.slice(93,117);
var blueSpec = ReflectanceSpectrumOffsetNorm.slice(51,67);

// Devide by number of pixels
var Red   = MathMEAN( redSpec )   || 0;  // (nm[681] - nm[631]) || 0;
var Green = MathMEAN( greenSpec ) || 0;  // (nm[580] - nm[530]) || 0;
var Blue  = MathMEAN( blueSpec )  || 0;  // (nm[475] - nm[437]) || 0;

var gamma = 1;

Red = Math.pow(Red,(1/gamma) ) * 255     || 0;
Green = Math.pow(Green,(1/gamma) ) * 255 || 0;
Blue = Math.pow(Blue,(1/gamma) ) * 255   || 0;


if(Red > 255 || Green > 255 || Blue > 255){

  var F_max = MathMAX([Red,Green,Blue]);
  
  Red *= (255/F_max);
  Green *= (255/F_max);
  Blue *= (255/F_max);
 
}


Red = MathROUND(Red,0);
Green = MathROUND(Green,0);
Blue = MathROUND(Blue,0);
  

Red = (Red < 0 || isNaN(Red)) ? 0 : Red;
Green = (Green < 0 || isNaN(Green)) ? 0 : Green;
Blue = (Blue < 0 || isNaN(Blue)) ? 0 : Blue;


// Start outputting spectra here
output.Color = [Red,Green,Blue];
output.Reflectance = 'rgb('+Red+','+Green+','+Blue+')';
output.Reflectance_Spectrum = ReflectanceSpectrumOffsetNorm.slice(51,225);
output.Red = Red;
output.Green = Green;
output.Blue = Blue;
output.original_trace = json.data_raw;

//Return data
return output;
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  "firmware_version": "0.457",
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  "app_os": "mac",
  "app_name": "PhotosynQ",
  "app_version": "0.3.3",
  "app_device": "x86-64",
  "ConsoleMacro": "180"
}
Sebastian kuhlgert square
Created by

Sebastian


Protocol connections:
15
Latest Update:
Sep 2016