Method for pre-identification of spectral overlaps within fluorescent dye and detector combinations used in flow cytometry
View Patent ↗This invention relates to the field of flow cytometry. It provides methods for identifying important dye-detector spectral overlaps for use in designing flow cytometry experiments. It further provides methods for quantifying the impact of spectral overlaps on dye selection and detector selection, the methods including the steps of: a) obtaining spectra of a dye; b) obtaining a laser configuration and an optical filter configuration of a detector; c) obtaining a spectrum yield value; and d) ranking the spectrum yield value.
1. A method for configuring a particle analysis mechanism by quantifying the impact of spectral overlaps on dye selection and detector selection comprising:
(a) obtaining a spectrum of a dye,
(b) obtaining a laser configuration and an optical filter configuration of a detector,
(c) obtaining a spectrum yield value,
(d) ranking the spectrum yield value,
(e) quantifying the impact of spectral overlaps on dye selection and detector selection, and
(f) configuring the particle analysis mechanism according to the results of the quantifying step.
2. The method according to claim 1 , wherein the spectrum is an excitation spectrum.
3. The method according to claim 1 , wherein the spectrum is an emission spectrum.
4. The method according to claim 1 , wherein the detector of step (b) is a component of a flow cytometer.
5. The method according to claim 1 , wherein the spectrum yield value is ranked in step (d) as minimal.
6. The method according to claim 1 , wherein the spectrum yield value is ranked in step (d) as substantial.
7. The method according to claim 1 , wherein the spectrum yield value in step (c) is obtained for a plurality of dyes.
8. The method according to claim 1 , wherein the spectrum yield value in step (c) is obtained for a plurality of detectors.
9. The method according to claim 1 , wherein the dye in step (a) is selected from the group consisting of fluorescein isothiocyanate, R-phycoerythrin, a tandem conjugate of sulforhodamine 101 acid chloride with R-phycoerythrin, a tandem conjugate of 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine -5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, enhanced green fluorescent protein, enhanced yellow fluorescent protein, DsRed, allophycocyanin, peridinin chlorophyll protein, coumarin, 3-(εcarboxypentyl)-3′-ethyl-oxacarbocyanine-6,6′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindocarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′, 3′-tetramethyl-3H-benz(e)indocarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl -3H-benz(e)indodicarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine -5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 2′-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-5′-bi-1H-benzimidazole 2-[4-(aminoiminomethyl) phenyl]-1H-indole-6-carboximidamide, Phenol, 4-[5-(4-methyl-1-piperazinyl) [2,5′-bi-1H-benzimidazol]-2′-yl]-, trihydrochloride chromomycin A3, mithramycin, Quinolinium, 1,1-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)- benzoxazolylidene)methyl]]-, tetraiodide, ethidium bromide, 7-aminoactinomycin D, acridine orange, Quinolinium, 1-1-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis [4-[(3-methyl-2 (3H)-benzothiazolylidene)methyl]]-, tetraiodide, Quinolinium, 4-[(3-methyl-2(3H)-benzothiazolylidene) methyl]-1-[3-(3-(trimethylammonio)propyl]-, diiodide, quinolinium, 1-methyl-4-((3-methyl-2(3H)-benzothiazolylidene)methyl), salt with 4-methylbenzenesulfonic acid (1:1), Quinolinium, 1,1′-[1,3-propanediylbis [(dimethylminio)-3,1-propanediyl]]bis[4-[3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-, tetraiodide, Quinolinium, 4-[3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-1-[3-(trimethylammonio) propyl]-, diiodide, propidium iodide, LDS 751, 2-[4-[bis(carboxymethyl)amino]-3-[2-[2-[bis(carboxymethyl)amino]-5-methylphenoxy]ethoxy]phenyl]-1H-indole-6-carboxylic acid, N-[2-[2-[2-[bis(carboxymethyl)amino]-5-(2,7-dichloro-6-hydroxy-3-oxo-3H-xanthen-9-yl) phenoxyl]ethoxy]-4-methylphenyl]-N-(carboxymethyl)-glycine, dichlorodihydrofluorescein diacetate, dihydrorhodamine SNARF, Y66F , Y66H, enhanced blue fluorescent protein, green fluorescent protein uv, enhanced cyan fluorescent protein, green fluorescent protein, AmCyan1, Y77W, S65A, S65C, S65L, S65T, ZsGreen1, ZsYellow1, DsRed2, DsRed monomer, AsRed2, mRFP1, HcRed1, monochlorobimane, calcein, cyanine, hydroxycoumarin, aminocoumarin, methoxycoumarin, acetic acid, [(3,6,8-trisulfo-1-pyrenyl)oxy]-, 1-hydrazide, trisodium salt, 1H-Benz[de]isoquinoline-5,8-disulfonic acid, 6-amino-2-[(hydrazinocarbonyl)aminol]-2,3-dihydro-1,3-dioxo-, dilithium salt, 6-[(7-nitro-2, 1,3-benzoxadiazol-4-yl)amino]-hexanoic acid, conjugates of 1-(ε-carboxypentyl) -1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, conjugates of allophycocyanin with 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5, 5′-disulfonate potassium salt N-hydroxysuccinimide ester, Red 613, fluorescein, 6-(5-and 6-fluoresceinyl-carboxamido)-hexanoic acid N-hydroxysuccinimide ester, 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid, tetramethylrhodamine-6-isothiocyanate, X-rhodamine, lissamine rhodamine B, sulforhodamine 101 acid chloride, conjugate of peridinin chlorophyll protein with 1-(ε-carboxypentyl) -1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz(e)indodicarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide este, and fluorescent derivatives thereof.
10. The method according to claim 1 , further comprising performing at least quantifying step (e) on a computer programmed with software to perform the quantifying step.
11. The method according to claim 1 , wherein the dye in step (a) comprises a compound selected from the group consisting of
fluorescent derivatives thereof.
12. A method for selecting dyes for use in flow cytometry comprising:
(a) obtaining a spectrum for each of a plurality of dyes in a form capable of being operated on by a digital computing device,
(b) obtaining a laser and optical filter detector configuration for at least one detector in a form capable of being operated on by a digital computing device,
(c) obtaining, via a digital computing device, for each of a plurality of combinations of a dye and a detector configuration for the at least one detector, a spectrum yield value,
(d) ranking the spectrum yield values,
(e) selecting a dye based upon the ranking of the spectrum yield values in step (d).
13. The method according to claim 12 , wherein the spectrum is an excitation spectrum.
14. The method according to claim 12 , wherein the spectrum is an emission spectrum.
15. The method according to claim 12 , wherein the detector of step (b) is a component of a flow cytometer.
16. The method according to claim 12 , wherein the spectrum yield value is ranked in step (d) as minimal.
17. The method according to claim 12 , wherein the spectrum yield value is ranked in step (d) as substantial.
18. The method according to claim 12 , wherein the spectrum yield value in step (c) is obtained for a plurality of dyes.
19. The method according to claim 12 , wherein the dye in step (a) is selected from the group consisting of fluorescein isothiocyanate, R-phycoerythrin, a tandem conjugate of sulforhodamine 101 acid chloride with R-phycoerythrin, a tandem conjugate of 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, enhanced green fluorescent protein, enhanced yellow fluorescent protein, DsRed, allophycocyanin, peridinin chlorophyll protein, coumarin, 3-(ε-carboxypentyl)-3′-ethyl-oxacarbocyanine-6,6′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindocarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz(e)indocarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz(e)indodicarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 2′-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-5′-bi-1H-benzimidazole, 2-[4-(aminoiminomethyl)phenyl]1-1H-indole-6-carboximidamide, Phenol, 4-[5-(4-methyl-1-piperazinyl)[2,5′-bi-1H-benzimidazo]-2′-yl]-, trihydrochloride chromomycin A3, mithramycin, Quinolinium, 1,1-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl ]]bis[4-[(3-methyl-2(3H)-benzoxazolylidene)methyl]]-, tetraiodide, ethidium bromide, 7-aminoactinomycin D, acridine orange, Quinolinium, 1-1-[1,3- propanediylbis[(dimethyliminio)-3,1-propanediyll]]bis [4-(3-methyl-2(3H)-benzothiazolylidene)methyl]]-, tetraiodide, Quinolinium, 4-[(3-methyl-2(3H)-benzothiazolylidene) methyl]-1-[3-(trimethylammonio)propyl]-, diiodide, quinolinium, 1-methyl-4-((3-methyl-2(3H)-benzothiazolylidene)methyl), salt with 4-methylbenzenesulfonic acid (1:1), Quinolinium, 1,1′-[1,3-propanediylbis [(dimethylminio)-3,1-propanediyl]]bis[4-3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-, tetraiodide, Quinolinium, 4-[3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-1-[3-(trimethylammonio) propyl]-, diiodide, propidium iodide, LDS 751, 2-[4-[bis(carboxymethyl)amino]-3-[2-[2-[bis(carboxymethyl)amin]-5-methylphenoxy]lethoxy]phenyl]-1H-indole-6-carboxylic acid, N-[2-[2-[2-[bis (carboxymethyl)amino]-5-(2,7-dichloro-6-hydroxy-3-oxo-3H-xanthen-9-yl) phenoxy]ethoxy]-4-methylphenyl]-N-(carboxymethyl)-glycine, dichlorodihydrofluorescein diacetate, dihydrorhodamine, SNARF, Y66F, Y66H, enhanced blue fluorescent protein, green fluorescent protein uv, enhanced cyan fluorescent protein, green fluorescent protein, AmCyan1, Y77W, S65A, S65C, S65L, S65T, ZsGreen1, ZsYellow1, DsRed2, DsRed monomer, AsRed2, mRFP1, HcRed1, monochlorobimane, calcein, cyanine, hydroxycoumarin, aminocoumarin, methoxycoumarin, acetic acid, [(3,6,8-trisulfo-1-pyrenyl)oxy]-, 1-hydrazide, trisodium salt, 1H-Benz[de]isoquinoline-5,8-disulfonic acid, 6-amino-2-[(hydrazinocarbonyl) amino]-2,3-dihydro-1,3-dioxo-, dilithium salt, 6-[(7-nitro-2, 1,3-benzoxadiazol-4-yl)aminol-hexanoic acid, conjugates of 1-(ε-carboxypentyl) -1-′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, conjugates of allophycocyanin with 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5, 5′-disulfonate potassium salt N-hydroxysuccinimide ester, Red 613, fluorescein, 6-(5-and 6-fluoresceinyl-carboxamido)-hexanoic acid N- hydroxysuccinimide ester, 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid, tetramethylrhodamine-6-isothiocyanate, X-rhodamine, lissamine rhodamine B, sulforhodamine 101 acid chloride, conjugate of peridinin chlorophyll protein with 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz(e)indodicarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, and fluorescent derivatives thereof.
20. The method according to claim 12 , wherein the dye in step (a) comprises a compound selected from the group consisting of
fluorescent derivatives thereof.
21. A method for selecting a detector for use in flow cytometry comprising:
(a) obtaining a spectrum of at least one dye in a form capable of being operated on by a dieital computing device,
(b) obtaining a laser and optical filter configuration for each of a plurality of detectors in a form capable of being operated on by a digital computing device,
(c) obtainin, via a digital computing device, for each of a plurality of combinations of the at least one dye and a detector configuration, a spectrum yield value,
(d) ranking the spectrum yield values,
(e) selecting a detector based upon ranking of the spectrum yield values in step (d) wherein the detector produces a maximum spectrum yield value in relation to the bandwidth of the optical filter.
22. The method according to claim 21 , wherein the spectrum is an excitation spectrum.
23. The method according to claim 21 , wherein the spectrum is an emission spectrum.
24. The method according to claim 21 , wherein the detector of step (b) is a component of a flow cytometer.
25. The method according to claim 21 , wherein the spectrum yield value in step (c) is obtained for a plurality of dyes.
26. The method according to claim 21 , wherein the dye in step (a) wherein the dye in step (a) is selected from the group consisting of fluorescein isothiocyanate, R-phycoerythrin, a tandem conjugate of sulforhodamine 101 acid chloride with R-phycoerythrin, a tandem conjugate of 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, enhanced green fluorescent protein, enhanced yellow fluorescent protein, DsRed, allophycocyanin, peridinin chlorophyll protein coumarin 3-(ε-carboxypentyl)-3′-ethyl -oxacarbocyanine-6,6′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl) -1′-ethyl-3,3,3′,3′-tetramethylindocarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz (e)indocarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindodicarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethyl -3H-benz(e)indodicarbocyanine-5,5′,7,7′-tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine -5,5′-disulfonate potassium salt N-hydroxysuccinimide ester, 2′-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-5′-bi-1H-benzimidazole, 2-[4-(aminoiminomethyl) phenyl]-1H-indole-6-carboximidamide, Phenol, 4-[5-(4-methyl-1-piperazinyl) [2,5′-bi-1H-benzimidazo]-2′-yl]-, trihydrochloride chromomycin A3, mithramycin, Quinolinium, 1,1-[1,3- propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)- benzoxazolylidene)methyl]]-, tetraiodide, ethidium bromide, 7-aminoactinomycin D, acridine orange, Quinolinium, 1-1-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediy]]bis[4-[(3-methyl-2(3H) -benzothiazolylidene)methyl]]-, tetraiodide, Quinolinium, 4-[(3-methyl-2(3H)-benzothiazolylidene) methyl]-1-[3-(trimethylammonio)propyl]-, diiodide, quinolinium, 1-methyl-4-((3-methyl-2(3H)-benzothiazolylidene)methyl), salt with 4-methylbenzenesulfonic acid (1:1), Quinolinium, 1,1′-[1,3-propanediylbis [(dimethylminio)-3,1-propanediyl]]bis[4-[3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-, tetraiodide, Quinolinium, 4-[3-(3-methyl-2(3H)-benzothiazolylidene)-1-propenyl]-1-[3-(trimethylammonio) propyl]-, diiodide, propidium iodide, LDS 751, 2-[4-[bis(carboxymethyl)amino]-3-[2-[2-[bis(carboxymethyl)amino]-5-methylphenoxy]ethoxyl]phenyl]-1H-indole-6-carboxylic acid, N-[2-[2-[2-[bis(carboxymethyl)amino]-5-(2,7-dichloro-6-hdroxy-3-oxo-3H-xanthen-9-yl) phenoxyl]ethoxy]-4-methylphenyl]-N-(carboxymethyl)-glycine, dichlorodihydrofluorescein diacetate, dihydrorhodamine, SNARF, Y66F , Y66H, enhanced blue fluorescent protein, green fluorescent protein uv, enhanced cyan fluorescent protein, green fluorescent protein, AmCyan1, Y77W, S65A, S65C, S65L, S65T, ZsGreen1, ZsYellow1, DsRed2, DsRed monomer, AsRed2, mRFP1, HcRed1, monochlorobimane, calcein, cyanine, hydroxycoumarin, aminocoumarin, methoxycoumarin, acetic acid, [(3,6,8-trisulfo-1-pyrenyl)oxy]-, 1-hydrazide, trisodium salt, 1H-Benz[de]lisoquinoline-5,8-disulfonic acid, 6-amino-2-[(hydrazinocarbonyl)aminol]-2,3-dihydro-1,3-dioxo-, dilithium salt, 6-[(7-nitro-2, 1,3-benzoxadiazol-4-yl)aminol-hexanoic acid, conjugates of 1-(ε-carboxypentyl) -1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5,5′-disulfonate potassium salt N-hydroxysuccinimide ester with R-phycoerythrin, conjugates of allophycocyanin with 1-(ε-carboxypentyl)-1′-ethyl-3,3,3′,3′-tetramethylindotricarbocyanine-5, 5′-disulfonate potassium salt N-hydroxysuccinimide ester, Red 613, fluorescein, 6-(5-and 6-fluoresceinyl-carboxamido)-hexanoic acid N-hydroxysuccinimide ester, 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid tetramethylrhodamine-6-isothiocyanate, X-rhodamine, lissamine rhodamine B, sulforhodamine 101 acid chloride, conjugate of peridinin chlorophyll protein with 1-(ε-carboxypentyl) -1′-ethyl-3,3,3′,3′-tetramethyl-3H-benz(e)indodicarbocyanine-5,5′,7,7′tetrasulfonate tripotassium salt N-hydroxysuccinimide ester, and fluorescent derivatives thereof.
27. The method according to claim 21 , further comprising performing the ranking step on the digital computing device programmed with software to perform the ranking step.
28. The method according to claim 21 , wherein the dye in step (a) comprises a compound selected from the group consisting of
fluorescent derivatives thereof.
29. A particle analysis mechanism, comprising:
a stimulation emission mechanism, emitting a stimulation emission having a spectrum;
a particle marked with a material responsive to the stimulation emission;
a particle response detector, having a configuration, detecting a response coming from the material;
the stimulation emission mechanism and the response detector having been selected based criteria determined according to the following:
obtaining the spectrum of the stimulation emission;
obtaining a spectrum yield based upon the stimulation emission spectrum and the configuration;
repeating the obtaining the spectrum and the obtaining the spectrum yield for a plurality of combinations of the emission spectrum and the configuration;
ranking the spectrum yields for the plurality of combinations; and,
selecting the stimulation emission mechanism and the response detector based upon the ranking of the spectrum yields for the different combinations.
30. The particle analysis mechanism of claim 29 further comprising:
the response detector configuration comprising a light source configuration and a filter configuration.
31. The particle analysis mechanism of claim 29 further comprising:
the response detector configuration comprising a light detector detecting light within a detection spectrum and a filter selected based upon the response detector detection spectrum.
32. The particle analysis mechanism of claim 29 comprising:
a flow cytometry particle analyzer.
33. The particle analysis mechanism of claim 29 further comprising:
the stimulation emission mechanism comprising a light source.
34. The particle analysis mechanism of claim 29 further comprising:
the material comprising a dye.
35. A particle analysis mechanism, comprising:
a stimulation emission mechanism, emitting a stimulation emission having a spectrum;
a particle marked with a material responsive to the stimulation emission;
a particle response detector, having a configuration, detecting a response coming from the material;
the material having been selected based on criteria determined according to the following:
obtaining the spectrum of the stimulation emission;
obtaining a spectrum yield based upon the stimulation emission spectrum and the material and the configuration;
repeating the obtaining the spectrum and the obtaining the spectrum yield for a plurality of combinations of the emission spectrum, the material and the configuration;
ranking the spectrum yields for the plurality of combinations; and, selecting the material based upon the ranking of the spectrum yields for the different combinations.
36. A particle analysis mechanism, comprising:
a stimulation emission mechanism, emitting a stimulation emission having a spectrum;
a particle marked with a material responsive to the stimulation emission;
a particle response detector, having a configuration, detecting a response coming from the material;
the response detector having been selected based criteria determined according to the following:
obtaining the spectrum of the stimulation emission;
obtaining a spectrum yield based upon the stimulation emission spectrum and the material and the configuration;
repeating the obtaining the spectrum and the obtaining the spectrum yield for a plurality of combinations of the emission spectrum, the material and the configuration;
ranking the spectrum yields for the plurality of combinations; and,
selecting the response detector based upon the ranking of the spectrum yields for the different combinations.
37. The particle analysis mechanism of claim 36 further comprising:
the response detector comprising a light detector detecting light within a detection spectrum and a filter selected based upon the response detector detection spectrum.
38. The particle analysis mechanism of claim 36 further comprising:
the response detector comprising a plurality of response detectors.