Method for improved selectivity in photo-activation and detection of molecular diagnostic agents

Claim 3

The method of claim 1 wherein the light sufficient to promote a simultaneous two-photon excitation of the photo-active molecular agent is a focused beam of light.

Claim 4

The method of claim 2 wherein the focused beam of light is focused laser light.

Claim 5

The method of claim 1 further including a step of treating the plant or animal tissue with at least one photo-active molecular agent before treating said particular volume of plant or animal tissue with light, so that the particular volume of the plant or animal tissue contains at least a portion of the at least one photo-active molecular agent.

Claim 6

The method of claim 4 wherein the at least one photo-active molecular agent is selected from the group consisting of psoralen, 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), 5-chloromethyl-8-methoxypsoralen (HMT), angelicin (isopsoralen), 5-methylangelicin (5-MIP), 3-carboxypsoralen, porphyrin, haematoporphyrin derivative (HPD), photofrin II, benzoporphyrin derivative (BPD), protoporphyrin IX (PpIX), dye haematoporphyrin ether (DHE), polyhaematoporphyrin esters (PHE), 13,17-N,N,N-dimethylethylethanolamine ester of protoporphyrin (PH1008), tetra(3-hydroxyphenyl)-porphyrin (3-THPP), tetraphenylporphyrin monosulfonate (TPPS1), tetraphenylporphyrin disulfonate (TPPS2a), dihaematoporphyrin ether, mesotetraphenylporphyrin, mesotetra(4N-methylpyridyl)porphyrin (T4MpyP), octa-(4-tert-butylphenyl)tetrapyrazinoporphyrazine (OPTP), phthalocyanine, tetra-(4-tert-butyl)phthalocyanin (t 4 -PcH 2 ), tetra-(4-tert-butyl)phthalocyanatomagnesium (t 4 -PcMg), chloroaluminum sulfonated phthalocyanine (CASPc), chloroaluminum phthalocyanine tetrasulfate (AlPcTS), mono-sulfonated aluminum phthalocyanine (AlSPc), di-sulfonated aluminum phthalocyanine (AlS2Pc), tri-sulfonated aluminum phthalocyanine (AlS3Pc), tetra-sulfonated aluminum phthalocyanine (Als4Pc), silicon phthalocyanine (SiPc IV), zinc II phthalocyanine (ZnPc), bis(di-isobutyl octadecylsiloxy)silicon 2,3-naphthalocyanine (isoBOSINC), germanium IV octabutoxyphthalocyanine (GePc), rhodamine 101 (Rh-101), rhodamine 110 (Rh-110), rhodamine 123 (Rh-123), rhodamine 19 (Rh-19), rhodamine 560 (Rh-560), rhodamine 575 (Rh-575), rhodamine 590 (Rh-590), rhodamine 610 (Rh-610), rhodamine 640 (Rh-640), rhodarnine 6G (Rh6G), rhodamine 700 (Rh-700), rhodamine 800 (Rh-800), rhodamine B (Rh-B), sulforhodamine 101, sulforhodamine 640, sulforhodamine B, coumarin 1, coumarin 2, coumarin 4, coumarin 6, coumarin 6H, coumarin 7, coumarin 30, coumarin 47, coumarin 102, coumarin 106, coumarin 120, coumarin 151, coumarin 152, coumarin 152A, coumarin 153, coumarin 311, coumarin 307, coumarin 314, coumarin 334, coumarin 337, coumarin 343, coumarin 440, coumarin 450, coumarin 456, coumarin 460, coumarin 461, coumarin 466, coumarin 478, coumarin 480, coumarin 481, coumarin 485, coumarin 490, coumarin 500, coumarin 503, coumarin 504, coumarin 510, coumarin 515, coumarin 519, coumarin 521, coumarin 522, coumarin 523, coumarin 535, coumarin 540, coumarin 540A, coumarin 548, 5-ethylamino-9-diethylaminobenzo a!phenoxazinium (EtNBA),5-ethyl-amino-9-diethylaminobenzo a!phenothiazinium (EtNBS), 5-ethylamino-9-diethylaminobenzo alphenoselenazinium (EtNBSe), chlorpromazine, chlorpromazine derivatives, chlorophyll derivatives, bacteriochlorophyll derivatives, metal-ligand complexes, tris(2,2'-bipyridine)ruthenium (II) dichloride (RuBPY), tris(2,2'-bipyridine)rhodium (II) dichloride (RhBPY), tris(2,2'-bipyridine)platinum (II) dichloride (PtBPY), pheophorbide a, merocyanine 540, vitamin D, 5-amino-laevulinic acid, photosan, chlorin e6, chlorin e6 ethylenediamide, mono-L-aspartyl chlorin e6, phenoxazine Nile blue derivatives, stilbene, stilbene derivatives, 4-(N-(2-hydroxyethyl)-N-methyl)-aminophenyl)-4'-(6-hydroxyhexylsulfonyl)stilbene (APSS), and standard biological dyes and stains.

Claim 7

The method of claim 4 wherein the at least one photo-active molecular agent is at least one biogenic photo-active molecular agent that is specific to a particular tissue within the particular volume of plant or animal tissue.

Claim 8

The method of claim 6 wherein the at least one biogenic photo-active molecular agent includes a segment selected from the group comprising nucleic acids, amino acids, proteins, antibodies, ligands, haptens, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, lipids, lipid receptors, lipid complexing agents, protein receptors, protein complexing agents, chelators, liposomes, fullerenes, crown ethers, cyclodextrins and encapsulating vehicles.

Claim 9

The method of claim 7 wherein the at least one biogenic photo-active molecular agent further includes a segment which is photo-activated when subject to light sufficient to promote a simultaneous two-photon excitation.

Claim 10

The method of claim 1 wherein the step of treating the particular volume of the plant or animal tissue with light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent contained in the particular volume of the plant or animal tissue includes the steps of: (a1) modulating light from a light source with a selected type of modulation, thereby producing a modulated light; and (a2) treating the particular volume of the plant or animal tissue with the modulated light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent contained therein; and wherein said method further includes the steps of: (f) demodulating said detected energy signal with said selected type of modulation; and (g) producing a demodulated energy signal which is characteristic of the particular volume of the plant or animal tissue.

Claim 11

The method of claim 9 wherein the step of demodulating the detected energy signal with the particular type of modulation includes demodulating the detected energy signal at a frequency which is twice that of said selected type of modulation, thereby detecting second harmonic of the selected type of modulation.

Claim 12

The method of claim 9 wherein the demodulated energy signal which is characteristic of the particular volume of the plant or animal tissue represents a change in lifetime of at least one photo-activated molecular agent present in the particular volume of the plant or animal tissue.

Claim 13

The method of claim 1 wherein the treating with light step includes: focusing a beam of light over a range of focal lengths so that a focal plane of the light beam extends to a location between a surface of the tissue and a point substantially beyond the tissue surface, whereby the treating step may extend to penetrate deep within the tissue.

Claim 14

The method of claim 1 wherein said method of identifying is for use in imaging said particular volume of plant and animal tissue.

Claim 15

The method of claim 13, wherein said detected energy signal is used in forming an image of said particular volume of plant or animal tissue.

Claim 16

The imaging method of claim 14 wherein the treating with light step includes: focusing a beam of light over a range of focal lengths so that a focal plane of the light beam extends to a location between a surface of the tissue and a point substantially beyond the tissue surface, whereby the treating step may extend to penetrate deep within the tissue, further including varying, the focal length position of the light beam within the tissue, so that said steps of photo-activating, detecting, and producing a detected energy signal occur along varying positions between the tissue surface and a position located substantially beyond the tissue surface, whereby said image is three dimensional.

Claim 17

The method of claim 1 wherein said at least one photo-active molecular agent is an endogenous agent.

Claim 18

The method of claim 1 wherein said at least one photo-active molecular agent is an exogenous agent.

Claim 19

The method of claim 1 wherein said at least one photo-active molecular agent includes endogenous agents selected from the group comprising aromatic amino acids, nucleic acids, proteins, cellular energy exchange stores such as adenosine triphosphate, enzymes, and hormones.

Claim 20

The method of claim 1 wherein said at least one photo-active molecular agent is coupled to a targeting agent before said step of treating the particular volume of plant or animal tissue with light, and wherein said targeting material is selected from the group comprising nucleic acids, amino acids, proteins, protein receptors, protein complexing agents, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, lipids, lipid receptors, lipid complexing agents, antibodies, ligands, haptens, complexing agents, chelators, encapsulating vehicles, liposomes, fullerenes, crown ethers, and cyclodextrins.

Claim 21

The method of claim 1 wherein the light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent is laser light.

Claim 22

The method of claim 20 wherein said laser light is produced by operating a laser to produce a pulsed output having a pulse repetition frequency above about 75 megahertz and a sub-nanosecond pulse duration.

Claim 23

The method of claim 21 including operating the laser to produce near-infrared light.

Claim 24

The method of claim 22 wherein the laser produces pulse energies of about 20 nanojoules.

Claim 25

The method of claim 21 wherein said detecting step comprises detecting emitted light that does not retrace an optical path of the incident light from the laser.

Claim 26

The method the claim 21 wherein said treating step further includes modulating the laser light; and wherein in one of the detecting step and the producing steps, a wavelength selection apparatus is used to filter energy emitted by said photo-activated agent.

Claim 27

The method of claim 21 wherein said treating and photo-activating steps produce emitted light which is from the molecular agent in the tissue and said production of emitted light is substantially synchronous with a modulation of the laser light.

Claim 30

The method of claim 27 wherein the material is selected from the group consisting of plant tissue and animal tissue.

Claim 31

The method of claim 27 wherein the light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent is laser light.

Claim 32

The method of claim 27 wherein the light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent is a focused beam of light.

Claim 33

The method of claim 30 wherein the focused beam of light is focused laser light.

Claim 34

The method of claim 27 further including a step of treating the material with at least one photo-active molecular agent before treating said particular volume of material with light, so that the particular volume of the material contains at least a portion of the at least one photo-active molecular agent.

Claim 35

The method of claim 32 wherein the at least one photo-active molecular agent is selected from the group consisting of psoralen, 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), 5-chloromethyl-8-methoxypsoralen (HMT), angelicin (isopsoralen), 5-methylangelicin (5-MIP), 3-carboxypsoralen, porphyrin, haematoporphyrin derivative (HPD), photofrin II, benzoporphyrin derivative (BPD), protoporphyrin IX (PpIX), dye haematoporphyrin ether (DHE), polyhaematoporphyrin esters (PHE), 13,17-N,N,N-dimethylethylethanolamine ester of protoporphyrin (PH1008), tetra(3-hydroxyphenyl)-porphyrin (3-THPP), tetraphenylporphyrin monosulfonate (TPPS1), tetraphenylporphyrin disulfonate (TPPS2a), dihaematoporphyrin ether, mesotetraphenylporphyrin, mesotetra(4N-methylpyridyl)porphyrin (T4MpyP), octa-(4-tert-(t 4 -PcH 2 ), tetra-(4-tert-butyl)phthalocyanatomagnesium (t 4 -PcMg), chloroaluminum sulfonated phthalocyanine (CASPc), chloroaluminum phthalocyanine tetrasulfate (AlPcTS), mono-sulfonated aluminum phthalocyanine (AlSPc), di-sulfonated aluminum phthalocyanine (AlS2Pc), tri-sulfonated aluminum phthalocyanine (AlS3Pc), tetra-sulfonated aluminum phthalocyanine (AlS4Pc), silicon phthalocyanine (SiPc IV), zinc II phthalocyanine (ZnPc), bis(di-isobutyl octadecylsiloxy)silicon 2,3-naphthalocyanine (isoBOSINC), germanium IV octabutoxyphthalocyanine (GePc), rhodamine 101 (Rh-101), rhodamine 110 (Rh-110), rhodamine 123 (Rh-123), rhodamine 19 (Rh-19), rhodamine 560 (Rh-560), rhodamine 575 (Rh-575), rhodamine 590 (Rh-590), rhodamine 610 (Rh-610), rhodamine 640 (Rh-640), rhodamine 6G (Rh6G), rhodamine 700 (Rh-700), rhodamine 800 (Rh-800), rhodamine B (Rh-B), sulforhodamine 101, sulforhodamine 640, sulforhodamine B, coumarin 1, coumarin 2, coumarin 4, coumarin 6, coumarin 6H, coumarin 7, coumarin 30, coumarin 47, coumarin 102, coumarin 106, coumarin 120, coumarin 151, coumarin 152, coumarin 152A, coumarin 153, coumarin 311, coumarin 307, coumarin 314, coumarin 334, coumarin 337, coumarin 343, coumarin 440, coumarin 450, coumarin 456, coumarin 460, coumarin 461, coumarin 466, coumarin 478, coumarin 480, coumarin 481, coumarin 485, coumarin 490, coumarin 500, coumarin 503, coumarin 504, coumarin 510, coumarin 515, coumarin 519, coumarin 521, coumarin 522, coumarin 523, coumarin 535, coumarin 540, coumarin 540A, coumarin 548, 5-ethylamino-9-diethylaminobenzo a!phenoxazinium (EtNBA), 5-ethyl-amino-9-diethylaminobenzo a!phenothiazinium (EtNBS), 5-ethylamino-9-diethylaminobenzo a!phenoselenazinium (EtNBSe), chlorpromazine, chlorpromazine derivatives, chlorophyll derivatives, bacteriochlorophyll derivatives, metal-ligand complexes, tris(2,2'-bipyridine)ruthenium (II) bacteriochlorophyll derivatives, metal-ligand complexes, tris(2,2'-bipyridine)ruthenium (II) dichloride (RuBPY), tris(2,2'-bipyridine)rhodium (II) dichloride (RhBPY), tris(2,2'-bipyridine)platinum (II) dichloride (PIBPY), pheophorbide a, merocyanine 540, vitamin D, 5-amino-laevulinic acid, photosan, chlorin e6, chlorin e6 ethylenediamide, mono-L-aspartyl chlorin e6, phenoxazine Nile blue derivatives, stilbene, stilbene derivatives, 4-(N-(2-hydroxyethyl)-N-methyl)-aminophenyl)-4'-(6-hydroxyhexylsulfonyl)stilbene (APSS), and standard biological dyes and stains.

Claim 36

The method of claim 32 wherein the at least one photo-active molecular agent is at least one biogenic photo-active molecular agent that is specific to a particular substance within the particular volume of material.

Claim 37

The method of claim 34 wherein the at least one biogenic photo-active molecular agent includes a segment selected from the group comprising nucleic acids, amino acids, proteins, antibodies, ligands, haptens, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, lipids, lipid receptors, lipid complexing agents, protein receptors, protein complexing agents, chelators, liposomes, fullerenes, crown ethers, cyclodextrins and encapsulating vehicles.

Claim 38

The method of claim 35 wherein the at least one biogenic photo-active molecular agent further includes a segment which is photo-activated when subject to light sufficient to promote a simultaneous two-photon excitation.

Claim 39

The method of claim 27 wherein the step of treating the particular volume of the material with light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent contained in the particular volume of the material includes the steps of: (a1) modulating light from a light source with a selected type of modulation, thereby producing a modulated light; and (a2) treating the particular volume of the material with the modulated light sufficient to promote a simultaneous two-photon excitation of the at least one photo-active molecular agent contained therein; and wherein the method further includes the steps of: (f) demodulating said detected energy signal with said selected type of modulation; and (g) producing a demodulated energy signal which is characteristic of the particular volume of the material.

Claim 40

The method of claim 37 wherein the step of demodulating the detected energy signal with the particular type of modulation includes demodulating the detected energy signal at a frequency which is twice that of the particular type of modulation, thereby detecting the second harmonic of the particular type of modulation.

Claim 41

The method of claim 37 wherein the demodulated energy signal which is characteristic of the particular volume of the material represents a change in lifetime of at least one photo-activated molecular agent present in the particular volume of the material.

Claim 42

The method of claim 27 wherein said detected energy signal is used in forming an image of said particular volume of materials.

Claim 43

The method of claim 27 wherein said at least one photo-active molecular agent includes endogenous agents selected from the group comprising aromatic amino acids, nucleic acids, proteins, cellular energy exchange stores such as adenosine triphosphate, enzymes, and hormones.

Claim 44

The method of claim 27 wherein said at least one photo-active molecular agent is coupled to a targeting agent before said step of treating the material with light, and wherein said targeting material is selected from the group comprising nucleic acids, amino acids, proteins, protein receptors, protein complexing agents, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, antibodies, ligands, haptens, complexing agents, chelators, encapsulating vehicles, liposomes, fullerenes, crown ethers, and cyclodextrins.

Claim 47

The method of claim 43 wherein said step of directing light includes generating near infra-red light using a pulsed laser operating at short pulse widths and a high pulse repetition rate, and focusing said laser light into said tissue.

Claim 48

The method of claim 43 wherein said step of controlling the location comprises varying the position of the confocal region relative to said specific feature of interest or varying the position of the specific feature of interest relative to a fixed confocal region.

Claim 49

The method of claim 43 wherein said method further includes modulating the light before it is incident on the tissue and demodulating the detected energy signal.

Claim 50

The method of claim 43 wherein said method causes simultaneous two-photon excitation at the confocal region.

Claim 51

The method of claim 47 wherein said detected energy signal is used in forming an image of said confocal region.

Claim 52

The method of claim 43 wherein said detected energy signal is used in forming an image of said confocal region.

Claim 55

The method of claim 50 further comprising the steps of demodulating the detected modulated energy signal.

Claim 56

The method of claim 50 wherein said method of identifying is for use in imaging said material.

Claim 57

The method of claim 52 wherein said detected modulated energy signal is used in forming an image of said material.

Claim 58

The method of claim 53 further comprising the steps of: demodulating the detected modulated energy signal; and recording said demodulated energy signal as a function of the location of said at least one photo-activated molecular agent.

Claim 59

The method of claim 50 wherein said material is selected from the group consisting of plant tissue and animal tissue.

Claim 60

The method of claim 55 wherein said animal tissue is located in the body of the animal.

Claim 61

The method of claim 50 wherein said light source is a laser.

Claim 62

The method of claim 50 further including a step of treating the material with at least one photo-active molecular agent before treating said material wilh light.

Claim 63

The method of claim 58 wherein said at least one photo-active molecular agent is coupled to a targeting agent before said step of treating the material with modulated light, and wherein said targeting material is selected from the group comprising nucleic acids, amino acids, proteins, protein receptors, protein complexing agents, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, lipids, lipid receptors, lipid complexing agents, antibodies, ligands, haptens, complexing agents, chelators, encapsulating vehicles, liposomes, fullerenes, crown ethers, and cyclodextrins.

Claim 64

The method of claim 58 wherein the at least one photo-active molecular agent is selected from the group comprised of psoralen, psoralen derivatives, porphyrin, porphyrin derivatives, hematoporphyrin derivatives, phthalocyanine, phthalocyanine derivatives, rhodamine, rhodamine derivatives, coumarin, coumarin derivatives, benzophenoxazine, benzophenoxazine derivatives, chlorpromazine, chlorpromazine derivatives, chlorophyll, chlorophyll derivatives, bacteriochlorophyll derivatives, metal-ligand complexes, pheophorbide a, merocyanine 540, vitamin D, 5-amino-laevulinic acid, photosan, chlorin e6, chlorin e6 ethylenediamide, mono-L-aspartyl chlorin e6, phenoxazine Nile blue, phenoxazine Nile blue derivatives, charge transfer agents, rediative transfer agents and standard biological dyes and stains.

Claim 65

The method of claim 60 wherein said psoralen derivatives include 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), 5-chloromethyl-8-methoxypsoralen (HMT), angelicin (isopsoralen), 5-methylangelicin (5-MIP), 3-carboxypsoralen, said porphyrin and haematoporphyrin derivatives include haematoporphyrin derivative (HPD), photofrin II, benzoporphyrin derivative (BPD), protoporphyrin IX (PpIX), dye haematoporphyrin ether (DHE), polyhaematoporphyrin esters (PHE), 13,17-N,N,N-dimethylethylethanolamine ester of protoporphyrin (PH1008), tetra(3-hydroxyphenyl)porphyrin (3-THPP), tetraphenylporphyrin monosulfonate (TPPS1), tetraphenylporphyrin disulfonate (TPPS2a), dihaematoporphyrin ether, mesotetraphenylporphyrin, mesotetra(4N-methylpyridyl)porphyrin (T4MpyP), octa-(4-tert-butylphenyl)tetrapyrazinoporphyrazine (OPTP), said phthalocyanine derivatives include tetra-(4-tert-butyl)phthalocyanine (t 4 -PcH 2 ), tetra-(4-tert-butyl)phthalocyanatomagnesium (t 4 -PcMg), chloroaluminum sulfonated phthalocyanine (CASPc), chloroaluminum phthalocyanine tetrasulfate (AlPcTS), monosulfonated aluminum phthalocyanine (AlSPc), di-sulfonated aluminum phthalocyanine (AlS2Pc), tri-sulfonated aluminum phthalocyanine (AlS3Pc), tetra-sulfonated aluminum phthalocyanine (AlS4Pc), silicon phthalocyanine (SiPc IV), zinc II phthalocyanine (ZnPc), bis(di-isobutyl octadecylsiloxy)silicon 2,3-naphthalocyanine (isoBOSINC), germanium IV octabutoxyphthalo-cyanine (GePc), said rhodamine derivatives including rhodamine 101 (Rh-101), rhodamine 110 (Rh-110), rhodamine 123 (Rh-123), rhodamine 19 (Rh-19), rhodamine 560 (Rh-560), rhodamine 575 (Rh-575), rhodamine 590 (Rh-590), rhodamine 610 (Rh-610), rhodamine 640 (Rh-640), rhodamine 6G (Rh-6G), rhodamine 700 (Rh-700), rhodamine 800 (Rh-800), rhodamine B (Rh-B), sulforhodamine 101, sulforhodamine 640, sulforhodamine B, said coumarin derivatives including coumarin 1, coumarin 2, coumarin 4, coumarin 6, coumarin 6H, coumarin 7, coumarin 30, coumarin 47, coumarin 102, coumarin 106, coumarin 120, coumarin 151, coumarin 152, coumarin 152A, coumarin 153, coumarin 311, coumarin 307, coumarin 314, coumarin 334, coumarin 337, coumarin 343, coumarin 440, coumarin 450, coumarin 456, coumarin 460, coumarin 461, coumarin 466, coumarin 478, coumarin 480, coumarin 481, coumarin 485, coumarin 490, coumarin 500, coumarin 503, coumarin 504, coumarin 510, coumarin 515, coumarin 519, coumarin 521, coumarin 522, coumarin 523, coumarin 535, coumarin 540, coumarin 540A, coumarin 548, said benzophemoxazine derivatives including 5-ethylamino-9-diethylaminobenzo a!phenoxazinium (EtNBA), 5-ethyl-amino-9-diethyl-aminobenzo a!phenothiazinium (EtNBS), 5-ethylamino-9-diethylaminobenzo a!pheno-selenazinium (EtNBSe), said chlorophyll and bacteriochlorophyll derivatives including bacteriochlorin a (BCA), said metal-ligand complexes including tris(2,2'-bipyridine)ruthenium (II) dichloride (RuBPY), tris(2,2'-bipyridine)rhodium (II) dichloride (RhBPY),tris(2,2'-bipyridine)platinum (II) dichloride (PtBPY), said phenoxazine Nile blue derivatives including phenoxazine dyes, said charge transfer and rediative transfer agents including stilbene, stilbene derivatives, 4-(N-(2-hydroxyethyl)-N-methyl)-aminophenyl)-4'-(6-hydroxyhexylsulfonyl)stilbene (APSS).

Claim 66

The method of claim 50 wherein said at least one photo-active molecular agent includes endogenous agents selected from the group comprising aromatic amino acids, nucleic acids, proteins, cellular energy exchange stores such as adenosine triphosphate, enzymes, and hormones.

Claim 69

The method of claim 63 further comprising the step of demodulating the detected modulated energy signal.

Claim 70

The method of claim 63 wherein said method of identifying is for use in imaging said tissue.

Claim 71

The method of claim 65 wherein said detected modulated energy signal is used in forming an image of said tissue.

Claim 72

The method of claim 66 further comprising the steps of: demodulating the detected modulated energy signal; and recording said demodulated energy signal.

Claim 73

The method of claim 67 further comprising the step of recording said demodulated energy signal as a function of the location of said confocal region.

Claim 74

The method of claim 63 wherein said tissue is located in the body of an animal.

Claim 75

The method of claim 63 wherein said light source is a laser.

Claim 76

The method of claim 63 further including a step of treating the material with at least one photo-active molecular agent before treating said material with light.

Claim 77

The method of claim 71 wherein the at least one photo-active molecular agent is selected from the group comprised of psoralen, psoralen derivatives, porphyrin, porphyrin derivatives, hematoporphyrin derivatives, phthalocyanine, phthalocyanine derivatives, rhodamine, rhodamine derivatives, coumarin, coumarin derivatives, benzophenoxazine, benzophenoxazine derivatives, chlorpromazine, chlorpromazine derivatives, chlorophyll, chlorophyll derivatives, bacteriochlorophyll derivatives, metal-ligand complexes, pheophorbide a, merocyanine 540, vitamin D, 5-amino-laevulinic acid, photosan, chlorin e6, chlorin e6 ethylenediamide, mono-L-aspartyl chlorin e6, phenoxazine Nile blue, phenoxazine Nile blue derivatives, charge transfer agents, rediative transfer agents and standard biological dyes and stains.

Claim 78

The method of claim 72 wherein said psoralen derivatives include 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), 5-chloromethyl-8-methoxypsoralen (HMT), angelicin (isopsoralen), 5-methylangelicin (5-MIP), 3-carboxypsoralen, said porphyrin and haematoporphyrin derivatives include haematoporphyrin derivative (HPD), photofrin II, benzoporphyrin derivative (BPD), protoporphyrin IX (PpIX), dye haematoporphyrin ether (DHE), polyhaematoporphyrin esters (PHE), 13,17-N,N,N-dimethylethylethanolamine ester of protoporphyrin (PH1008), tetra(3-hydroxyphenyl)porphyrin (3-TIPP), tetraphenylporphyrin monosulfonate (TPPS1), tetraphenylporphyrin disulfonate (kTPPS2a), dihaematoporphyrin ether, mesotetraphenylporphyrin, mesotetra(4N-methylpyridyl)porphyrin (T4MpyP), octa-(4-tert-butylphenyl)tetrapyrazinoporphyrazine (OPTP), said phthalocyanine derivatives include tetra-(4-tert-butyl)phthalocyanine (t 4 -PcH 2 ), tetra-(4-tert-butyl)phthalocyanatomagnesium (t 4 -PcMg), chloroaluminum sulfonated phthalocyanine (CASPc), chloroaluminum phthalocyanine tetrasulfate (AlPcTS), mono-sulfonated aluminum phthalocyanine (AlSPc), di-sulfonated aluminum phthalocyanine (AlS2Pc), tri-sulfonated aluminum phthalocyanine (AlS3Pc), tetra-sulfonated aluminum phthalocyanine (AlS4Pc), silicon phthalocyanine (SiPc IV), zinc II phthalocyanine (ZnPc), bis(di-isobutyl octadecylsiloxy)silicon 2,3-naphthalocyanine (isoBOSINC), germanium IV octabutoxyphthalo-cyanine (GePc), said rhodamine derivatives including rhodamine 101 (Rh-101), rhodamine 110 (Rh-110), rhodamine 123 (Rh-123), rhodamine 19 (Rh-19), rhodamine 560 (Rh-560), rhodamine 575 (Rh-575), rhodamine 590 (Rh-590), rhodamine 610 (Rh-610), rhodamine 640 (Rh-640), rhodamine 6G (Rh-6G), rhodamine 700 (Rh-700), rhodamine 800 (Rh-800), rhodamine B (Rh-B), sulforhodamine 101, sulforhodamine 640, sulforhodamine B, said coumarin derivatives including coumarin 1, coumarin 2, coumarin 4, coumarin 6, coumarin 6H coumarin 7, coumarin 30, coumarin 47, coumarin 102, coumarin 106, coumarin 120, coumarin 151, coumarin 152, coumarin 152A, coumarin 153, coumarin 311, coumarin 307, coumarin 314, coumarin 334, coumarin 337, coumarin 343, coumarin 440, coumarin 450, coumarin 456, coumarin 460, coumarin 461, coumarin 466, coumarin 478, coumarin 480, coumarin 481, coumarin 485, coumarin 490, coumarin 500, coumarin 503, coumarin 504, coumarin 510, coumarin 515, coumarin 519, coumarin 521, coumarin 522, coumarin 523, coumarin 535, coumarin 540, coumarin 540A, coumarin 548, said benzophemoxazine derivatives including 5-ethylamino-9-diethylaminobenzo a!phenoxazinium (EtNBA), 5-ethyl-amino-9-diethyl-aminobenzo a!phenothiazinium (EtNBS), 5-ethylamino-9-diethylaminobenzo a!pheno-selenazinium (EtNBSe), said chlorophyll and bacteriochlorophyll derivatives including bacteriochlorin a (BCA), said metal-ligand complexes including tris(2,2'-bipyridine)ruthenium (II) dichloride (RuBPY),tris(2,2'-bipyridine)rhodium (II) dichloride (RhBPY), tris(2,2'-bipyridine)platinum (II) dichloride (PtBPY), said phenoxazine Nile blue derivatives including phenoxazine dyes, said charge transfer and rediative transfer agents including stilbene, stilbene derivatives, 4-(N-(2-hydroxyethyl)-N-methyl)-aminophenyl)-4'-(6-hydroxyhexylsulfonyl)stilbene (APSS).

Claim 79

The method of claim 71 wherein said at least one photo-active molecular agent is coupled to a targeting agent before said step of directing the light into the tissue, and wherein said targeting material is selected from the group comprising nucleic acids, amino acids, proteins, protein receptors, protein complexing agents, carbohydrates, carbohydrate receptors, carbohydrate complexing agents, lipids, lipid receptors, lipid compleming agents, antibodies, ligands, haptens, complexing agents, chelators, encapsulating vehicles, liposomes, fullerenes, crown ethers, and cyclodextrins.

Claim 80

The method of claim 63 wherein said at least one photo-active molecular agent includes endogenous agents selected from the group comprising aromatic amino acids, nucleic acids, proteins, cellular energy exchange stores such as adenosine triphosphate, enzymes, and hormones.

Claim 83

The method of claim 76 wherein said method for diagnostic identification for use in imaging said tissue.

Claim 84

The method of claim 76 further including modulating said laser beam; wherein said photo-activated agent is caused to emit modulated energy, wherein said detecting step includes detecting modulated energy, wherein said producing step produces a detected modulated energy signal; and using said detected modulated energy signal to form an image of any said features of interest in said tissue.

Claim 85

The method of claim 76 wherein said photo-activating step causes fluorescence substantially only at any said feature of interest at said confocal region.