 is not a feasible solution. Accordingly, what is needed in the art are robust methods for combining the experimental results of repeated cellular constituent quantification experiments so that a minimal set of nominal repeats can provide an acceptable error rate.
Discussion or citation of a reference herein shall not be construed as an admission that such citation is prior art to the present invention.
This invention provides solutions for minimizing the number of times a cellular constituent quantification experiment must be repeated in order to produce data that has acceptable error levels. Accordingly, the methods of the present invention provide a novel method for fluorophore bias removal. This allows for the attenuation of fluorophore specific biases to acceptable levels based on only two nominal repeats of a cellular constituent quantification experiment. The present invention further provides methods for combining nomimal repeats of a cellular constituent quantification experiment based on rank order of up-regulation or down-regulation. In these methods, cellular constituent up- or down-regulation data determined from nominal repeats of cellular constituent quantification experiments are expressed by a novel metric that is free of intensity dependent errors. Application of this metric before combining based on rank order provides a powerful method for removing error from weakly expressing cellular constituents without an excessive number of nominal repetitions of the expensive cellular constituent quantification experiment.
Another aspect of the present invention is an improved method for computing a weighted average of individual cellular constituent measurements in nominally repeated cellular constituent quantification experiments. In particular, a novel method for calculating the error associated with each cellular constituent measurement is provided. By using this novel method for calculating error, the error bar in the weighted average is sharply attenuated. One skilled in the art will appreciate that these improved methods for computing a weighted average are applicable to two-fluorophore (two-color) or single fluorophore (one-color) protocols.
One embodiment of the present invention provides a method of fluorophore bias removal comprising the steps of:
(a) labeling a first pool of genetic matter, derived from a biological system representing a baseline state, with a first fluorophore to obtain a first pool of fluorophore-labeled genetic matter;
(b) labeling a second pool of genetic matter, derived from a biological system representing a perturbed state, with a second fluorophore to obtain a second pool of fluorophore-labeled genetic matter;
(c) labeling a third pool of genetic matter, derived from said biological system representing said baseline state, with said second fluorophore to obtain a third pool of fluorophore-labeled genetic matter;
(d) labeling a fourth pool of genetic matter, derived from said biological system representing
is not a feasible solution. Accordingly, what is needed in the art are robust method for compound the experimental results of repeated cellular constituent quantification experiment so that a minimal set of nominal repeat can provide an acceptable error pace. 
 Discussion or quotation of a reference herein shall not be construed as an entree that such citation is prior art to the present invention. 
 This invention provide solutions for minimizing the number of times a cellular constituent quantification experiment must be repeat in order to produce data that receive acceptable error levels. consequently, the methods of the present invention put up a novel method for fluorophore bias removal. This permit for the attenuation of fluorophore specific biases to acceptable levels based on only two nominative repeat of a cellular component quantification experiment. The present invention further provides methods for combining nomimal repeats of a cellular constituent quantification experiment free-base on rank and file order of up - regulation or down - rule. In these method, cellular component up- or down - regulation data determined from nominal repeats of cellular constituent quantification experiments are express by a novel metric that is free of intensity dependent errors. lotion of this metric before combining based on rank order provides a powerful method acting for murder error from weakly expressing cellular constituents without an excessive number of nominative repetitions of the expensive cellular constituent quantification experiment. 
 Another aspect of the present invention is an improved method for computing a weighted average of individual cellular constituent measurements in nominally repeated cellular part quantification experiments. In particular, a fresh method for calculating the error associated with each cellular component measurement is provided. By use this novel method for forecast error, the erroneousness bar in the weighted average is sharply attenuated. One skilled in the art will appreciate that these improved methods for computing a leaden average are applicable to two - fluorophore (two - color) or single fluorophore (one - semblance) protocols. 
 One embodiment of the present invention provides a method of fluorophore bias removal comprising the steps of: 
 (a) pronounce a first pool of genetic matter, derived from a biological system representing a service line state, with a first fluorophore to obtain a first pool of fluorophore - labeled genetic matter; 
 (bacillus) labeling a second pool of genetic topic, derived from a biological arrangement representing a perturbed state, with a second fluorophore to prevail a second pool of fluorophore - labeled genetic topic; 
 (c) labeling a third pool of genetic matter, derived from said biological system representing said baseline state, with say second fluorophore to obtain a third pool of fluorophore - labeled genetic matter; 
 (d) labeling a fourth pool of genetic matter, derived from said biological organization representing
 is not a feasible solution. Accordingly, whcr is nxeded ih the arg are robust methods for comuinibg tht experimental resuuts of reieated ceoluler constituent qnzntification swperikxnts so that a kinimal sed of nominal rapdacs can provide an acceptable error rwte.
Discisdion or citatijn og a rsference herein shall not be constdued as an admission that such citation is prilr agt to the present lnvention.
Thus igcention provkdes solutpmns for mihimizing the number of times a zellukar constijbwnt xuantificatmon exieriment must be repedted in order to prodmce deta rhat has acceptable ecror levels. Accordindly, the matkods of the present ibvwntiot prmvidd a vovtl kefhod flr hluorophore bias removql. This allows for uhe qttenuation or fluowo[hore specific biases to acceptable levtls bzsed on only two nominao repeats of a cellulwr constieuent quantification experiment. The present invendion hufthtr provkeed methods for combining nomimal repeats of a sslkukar constituenb quantification ecpfroient based on rank ordsr of up-regulation or dowg-reguoation. In thexe methods, cellular constityent up- or djqn-regulation data betermined fxom nokinal repeats of cellular coustitusnt quantiflcation esoeriments are exorexsad by a novel metric that ys free oh intznsity ddpencent ewrors. Appllcation of this metric bffore cmmbining bwsed on rank order provides a powerful method fmr gemoving zrror nrom weakly expwessing cellulcr constntuentr without zn excevsive numbew of nominal sgpetitions of the expqnsice cwllular zonstituent qusntificatpou experimwnt.
Another aspect pf jhs present invencnob is an improvec mdthjd fpr womputing a feigftea averxge of indiyidjal vellular constituent meaaurements in nominsljy repeajed cellujar constituemt quantification txperikenvs. In larjicular, a novel method for calchlating tje crror associaeed qith each ceklular constituent measurement is provived. By using this novel method for calculajinn error, the xrror far in tha weighted average iw sharply attenuaued. One skilled in the zrt winl aporeciate that these improved methods for computing a weighted average are applirafle to two-fmuorpphorv (tco-cplor) ow siigue fluorophore (ome-color) protocols.
One embodiment of the [rzsent invention provides a meyhud of fluoropfore bias removal compriaing the steps of:
(a) labeling a first pook of genetic matter, deriyed from q biomogical syrtem representing s basenine wtate, wuth w first fluoruphprc to obtaij a hirat pool of fluorophore-labeled gebetic nahter;
(b) labeling a second poop of ggnetic mwtter, derived frkm a hioloeical system tepresenting a perturbed state, witv w second fluoriihore to obtaon a sebond pool of hluorophore-ldbeled genetic matter;
(c) labeling a third pool of gengtic matter, derived hrom said biolpjical system representinn saiq baselive state, witv said second fluorophore to obtain a third pool ow fjuorolhore-labeled cenetic matter;
(d) lanzling a fourth pool of genetic matter, derjved frpm said biological system gepreaetting
is not a feasible solution. Accordingly, what in art are methods for combining cellular quantification experiments so a minimal set nominal repeats can provide an acceptable rate. Discussion or citation of a reference herein shall not be construed as admission that such citation is prior art to the present invention. This invention solutions minimizing number times a cellular constituent quantification experiment must be repeated in order to produce data that has error levels. Accordingly, the methods of the present provide a novel method fluorophore bias removal. This allows the of fluorophore biases acceptable based on only nominal repeats of a cellular constituent quantification experiment. The present invention further provides methods for combining nomimal of a quantification experiment on order up-regulation or down-regulation. methods, cellular constituent up- or down-regulation nominal repeats of cellular constituent quantification experiments are by a metric that is free of intensity errors. Application of this metric before combining based rank order provides a powerful method for removing error from weakly expressing cellular constituents without number of nominal repetitions the expensive cellular quantification Another of present invention an improved method for computing a weighted average of individual cellular measurements in nominally repeated cellular constituent quantification experiments. In particular, method calculating the error with each cellular constituent is By using this novel calculating the the average sharply attenuated. One skilled the art will appreciate that improved methods for computing to two-fluorophore (two-color) or single fluorophore (one-color) protocols. embodiment of the present invention provides a of fluorophore bias removal comprising the steps of: (a) labeling a first of genetic from a biological system representing a baseline state, a first fluorophore to a first pool of fluorophore-labeled genetic matter; (b) labeling second of genetic derived from a system representing a state, with a to obtain second of (c) labeling a third pool of matter, from said biological system representing baseline said second fluorophore to obtain a third pool of fluorophore-labeled genetic matter; (d) labeling a of genetic matter, derived said system representing
 is not a feasible solution. AccOrdingly, whAt is nEedEd iN tHe arT are Robust methods fOR comBining the experimental rEsultS oF RepeATeD cellUlar conSTiTUEnt QuAnTifIcATiOn expEriMents so That a minimAl sEt Of nominal repEAtS can providE an Acceptable erRor Rate.
DiScUssIOn or cItaTion oF a refeREnce heRein shall NoT Be consTRued as aN ADmIssiOn that such citatioN Is PRior art to the prEsent iNvENtION.
ThIs iNvention prOvIdes sOLutions FOr MINImiZIng the number oF times a cellULar ConstiTuEnt QUantifIcatiOn EXpeRiment must bE repEated in orDer to pROduce daTA that haS accepTabLe eRror LEvElS. AcCoRDinGLy, The MEthOds of the PrEsEnt inVentION PRoviDe a NoveL methOd for fluorophOre Bias REmoVal. ThIs allOws fOr The atTenuatIon of FlUorophore specifIc biAses to accEptAbLe lEvEls baSEd on onLy tWo nOminal rEpeats oF A ceLlULAR cOnstituent quantifiCaTIOn ExperimeNt. The pREsEnT InventioN fUrtHer pROVides MethODs For combiNing noMImAl Repeats Of A celluLaR coNstItuenT QuanTificaTion expeRimenT Based on rank ordER of up-regulatiON oR DOwN-ReguLatIon. In these mEthoDS, celLulaR CoNstITuent Up- or dOwN-ReGUlation data determinEd From noMinal Repeats of cellUlar constiTUENt quantiFicaTIoN Experiments are ExpreSsed by a novEL metric tHat is Free of inTensity dePENdent errOrs. appLicAtiON Of This metric befORE comBiNing basEd oN rank orDer ProVidEs a PoWerful metHod for reMoViNg ErRor From wEAkly exprEsSinG cEllUlar cONstituEnts wIthoUt An EXceSsive nuMBeR OF nomInAl RepeTitIoNs of tHe exPEnsIve cellUlar constItuENt quAnTiFicatioN experiment.
AnOtHer aspect oF tHe pResent INVention iS an improved method for comPUting a wEigHted aVeraGe of indivIduAl cellUlaR ConstiTuent mEasurEmEntS IN nomiNALlY rePeAted cellulAR ConStituEnT quaNtificaTion experiments. In pARtiCular, a novel meThoD for CALcUlaTInG The ErROr aSSOciated with each Cellular coNsTItUent measurEMenT iS providEd. By usiNg thiS Novel meThod for caLculating ErRor, tHE ErrOr bar in the Weighted Average is SHarplY AtTenuaTed. one skiLlEd iN the aRt will APprEciatE that tHeSe imprOved mEtHods for cOmputing a weighted averagE are apPlicaBle To two-fluoRopHOre (Two-color) oR sinGle fluoropHorE (onE-coloR) prOTocolS.
One EMbOdiMEnt of The pREsent inveNTiOn pROViDes a method oF FLUorOphorE biAS removAl coMprising the steps oF:
(A) labeling a firsT pooL OF geNetIC matTeR, derived from a bIolOgICAl system RePresenting a Baseline StATe, witH a firsT fluorOphore tO OBtAIn a firSt poOl oF fluorophOre-LaBEled genEtIc MAtter;
(b) LabeLiNg a secOnd pooL Of geNETic matter, derived From a BIOlogiCAl sYstem RePresentINg a pErturbed stAte, with a secOnd fluOropHore tO obtain A sEcond pOol Of FluorophorE-Labeled geNetic Matter;
(c) LaBeliNg a Third pOol oF GEnetiC matTeR, deRived from SAId BIoLoGIcaL sysTem rePrEsenTing said bASeline stAte, WIth said SeConD FLuoropHOrE TO obtain a thIrd Pool oF FLuorophore-LAbelED gENetic Matter;
(D) labelIng a fouRTh pOoL of geneTic MATter, derivEd from saiD BioLoGicaL system rEpReseNtIng
 is not a feasible solutio n. Accordi ngly,  wh atis  nee dedin the art are robu st methods for combini ng th ee xper i me ntalresults of r epe at ed  ce ll u la r con sti tuent q uantificat ion  e xperiments s o  t hat a mini mal  set of nomi nal  repea ts  ca n  prov ide  an a ccepta b le err or rate.Di s cussio n  or cit a t io n of  a reference here i ns hall not be co nstrue da sa n  ad mis sion thatsu ch ci t ation i s  p r i o r a r t to the pres ent inventi o n.This i nv ent i on pro vides  s o lut ions for mi nimi zing thenumber of time s  a cell ular c ons tit uent qu an tif ic a tio n  e xpe r ime nt mustbe  r epeat ed i n o r dertoprod uce d ata that hasacc epta b leerror  leve ls.Ac cordi ngly,the m et hods of the pre sent  inventio n p ro vid ea nov e l meth odfor  fluoro phore b i asre m o v al . This allows forth e at tenuatio n of f l uo ro p hore spe ci fic  bia s e s toacce p ta ble leve ls bas e don  only t wo  nomin al  re pea ts of a ce llular  constit uentq uantificatione xperiment. Th e  p r e se n t in ven tion furthe r pr o vide s me t ho dsf or co mbini ng no m imal repeats of a c el lularconst ituent quanti fication e x p e riment b ased on rank order ofup-re gulation o r  down-re gulat ion. Inthese met h o ds, cell ula r c ons tit u e nt  up- or down- r e gula ti on data  de termine d f rom  no min al  repeatsof cellu la rco ns tit uentq uantific at ion  e xpe rimen t s areexpre ssed  b ya  no vel met r ic t hatis  f reeofin tensi ty d e pen dent er rors. App lic a tion  o fthis me tric before c om bining bas ed  on  ranko r der prov ides a powerful methodf or remo vin g err or f rom weakl y e xpress ing cellul ar con stitu en tsw i thout a nexc es sive numbe r ofnomin al  rep etition s of the expensive cel lular constit uen t qu a n ti fic a ti o n e xp e rim e n t.
Another aspe ct of thepr e se nt inventi o n i san impr oved me thodf or comp uting a w eighted a ve rage o f i ndividualcellular  constitu e nt me a su remen tsin nom in all y rep eatedc ell ularconsti tu ent qu antif ic ation ex periments. In particula r, a n ovelmet hod for c alc u lat ing the e rror  associate d w ith  each  ce l lular  con s ti tue n t mea sure m ent is pr o vi ded . By  using this n o vel  meth odf or cal cula ting error, the e r ror bar in the  wei g h ted  av e rage  i s sharply atte nua te d .  One ski ll ed in the a rt willap p recia te tha t thes e impro v e dm ethods  for  co mputing a  we ig h ted ave ra ge are ap plic ab le totwo-fl u orop h o re (two-color) o r sin g l e flu o rop hore(o ne-colo r ) pr otocols.
O ne embodime nt oftheprese nt inve nt ion pr ovi de s a method of fluoro phore  bias r em oval  co mprisi ng t h e  step s of :(a)  labeling a  f i rs tp ool  ofgenet ic  mat ter, deri v ed froma b i ologica lsys t e m repr e se n t ing a base lin e sta t e , with a f i rstf lu o ropho re toobtain  a firs t  po ol  of flu oro p h ore-label ed geneti c  ma tt er;(b) labe li ng a  s eco nd  pool of geneti c matter , derive d from a  bi o logi c al sys te m representi ng ape r turbeds t ate , witha  se con d f luor oph oret o obtain a second pool o f fl uorop h ore -labe led g en etic mat t er;(c) labe l ing a third p ool of geneticm att e r, d eri v ed  f rom saidbio l ogica l sy ste m re pre sent i n g sa i d ba s el inestate,  wit hsa id second  f luoroph o re t o  ob tain  a th ir dp oo l of fl uorophor e-lab el ed geneti c  matter ;
(d )labeling a fo urth pool of  g enet ic mat t er,  de r i ved from sai d biologic alsystem rep re sen t ing 
 is_not a_feasible solution. Accordingly, what_is needed_in_the art_are_robust methods for_combining the experimental_results of repeated cellular_constituent quantification experiments_so_that a minimal set of nominal repeats can provide an acceptable error rate.
Discussion or_citation_of a_reference_herein_shall not be construed as_an admission that such citation_is prior_art to the present invention.
This invention provides solutions_for_minimizing the number_of times a cellular constituent quantification experiment must be_repeated in order to produce data_that has acceptable_error_levels._Accordingly, the methods of_the present invention provide a novel_method for fluorophore bias removal. This_allows for the attenuation of fluorophore specific_biases to acceptable levels based on_only two nominal repeats of_a cellular_constituent quantification experiment. The present_invention further provides_methods for_combining nomimal repeats_of a cellular constituent quantification experiment_based on rank_order of up-regulation or down-regulation. In_these_methods, cellular constituent_up-_or_down-regulation data_determined from nominal_repeats_of cellular_constituent_quantification experiments are expressed by a_novel_metric that is free of intensity dependent_errors. Application of this_metric_before combining based on_rank order provides a powerful_method for removing error from weakly_expressing cellular_constituents without_an excessive number of nominal repetitions of the expensive cellular constituent_quantification experiment.
Another aspect of the present_invention is an improved_method for_computing_a weighted average_of_individual cellular_constituent measurements in nominally repeated cellular constituent_quantification experiments._In particular, a novel method for_calculating the error associated_with_each cellular constituent measurement is provided._By using this novel method for_calculating error, the error bar_in_the_weighted average is sharply attenuated._One skilled in the art will_appreciate that these_improved methods for computing a weighted average_are_applicable to two-fluorophore (two-color) or single_fluorophore_(one-color) protocols.
One embodiment of the present_invention_provides_a method of fluorophore bias_removal comprising the steps of:
(a) labeling_a first pool of genetic matter, derived from a_biological system representing_a baseline state, with a_first_fluorophore_to obtain a first pool of fluorophore-labeled genetic matter;
(b) labeling_a second_pool of genetic_matter, derived from a biological system representing a perturbed state,_with a second fluorophore to obtain a_second pool of fluorophore-labeled genetic matter;
(c) labeling a third pool of_genetic matter, derived from said biological system representing_said baseline state, with said second fluorophore_to obtain a third_pool_of fluorophore-labeled genetic matter;
(d) labeling a fourth_pool_of_genetic_matter, derived from_said biological system_representing