US5109429A — Apparatus and method for analyses of biological s…

Claim 1

Independent Claim 152 words

A kit for the quantitation of cell nuclei comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference cell area containing a reference means for staining, the reference means having predetermined physical characteristics which are detectable after staining; and one or more containers of stain material; the stain material including stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers being in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 2

Dependent Claim 152 words

A kit for the quantitation of cell nuclei comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference cell area containing a reference means for staining, the reference means having predetermined physical characteristics which are detectable after staining; and one or more containers of stain material; the stain material including stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers being in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 3

Dependent Claim 16 words

A kit as recited in claim 1 wherein the kit further contains a rinse sulfonating agent.

Claim 4

Dependent Claim 17 words

A kit as recited in claim 1 wherein the reference means includes a monoclonal antibody associated therewith.

Claim 5

Dependent Claim 47 words

A kit as recited in claim 2 wherein the stain material includes Azure A in an effective amount to provide an aqueous solution with a concentration of Azure A in a range from about 4.9 to about 5.1 mg/ml when mixed with a 0.1N HCl aqueous solution.

Claim 6

Dependent Claim 17 words

A kit as recited in claim 4 wherein the stain material further includes a stain sulfonating agent.

Claim 7

Dependent Claim 14 words

A kit as recited in claim 3 wherein the stain includes an enzyme stain.

Claim 8

Dependent Claim 27 words

A kit as recited in claim 5 wherein the rinse sulfonating agent is K 2 S 2 O 5 or Na 2 S 2 O 5 .

Claim 9

Dependent Claim 27 words

A kit as recited in claim 7 wherein the stain sulfonating agent is K 2 S 2 O 5 or Na 2 S 2 O 5 .

Claim 10

Dependent Claim 35 words

A kit as recited in claim 1 wherein the stain is selected from the group consisting of Azure A, congo red, pararosanilin, hematoxylin, eosin, methyl green, diaminobenzidine, alkaline phosphatase, methylene blue, giemsa and mixtures thereof.

Claim 11

Dependent Claim 46 words

A kit as recited in claim 1 wherein the microscopic slide further includes an optical density reference area, the optical density reference area including a material having a predetermined known optical density to calibrate the slide with an instrument being used to study the specimen cells.

Claim 12

Dependent Claim 46 words

A kit as recited in claim 4 wherein the microscopic slide further includes an optical density reference area, the optical density reference area including a material having a predetermined known optical density to calibrate the slide with an instrument being used to study the specimen cells.

Claim 13

Dependent Claim 180 words

A kit for the quantitation of nuclear DNA by light microscopy, the kit comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference area containing a reference means for staining, the reference means having physical characteristics which permit the reference means to associate with a stain proportionally to an association of the stain with DNA; one or more containers of stain material, the stain material including a stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers selected from the group consisting of Azure A, pararosanilin, and mixtures thereof in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 14

Dependent Claim 180 words

A kit for the quantitation of nuclear DNA by light microscopy, the kit comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference area containing a reference means for staining, the reference means having physical characteristics which permit the reference means to associate with a stain proportionally to an association of the stain with DNA; one or more containers of stain material, the stain material including a stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers selected from the group consisting of Azure A, pararosanilin, and mixtures thereof in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 15

Dependent Claim 13 words

A kit as recited in claim 12 further comprising a rinse sulfonating agent.

Claim 16

Dependent Claim 51 words

A kit as recited in claim 13 wherein the stain material includes Azure A in an effective amount to provide an aqueous solution with concentration of Azure A in a range of from about 4.9 to about 5.1 mg/ml when the Azure A is mixed with a 0.1N HCl aqueous solution.

Claim 17

Dependent Claim 17 words

A kit as recited in claim 14 wherein the stain material further includes a stain sulfonating agent.

Claim 18

Dependent Claim 27 words

A kit as recited in claim 14 wherein the rinse sulfonating agent is K 2 S 2 O 5 or Na 2 S 2 O 5 .

Claim 19

Dependent Claim 46 words

A kit as recited in claim 12 wherein the microscopic slide further includes an optical density reference area, the optical density reference area including a material having a predetermined known optical density to calibrate the slide with an instrument being used to study the specimen cells.

Claim 20

Dependent Claim 46 words

A kit as recited in claim 14 wherein the microscopic slide further includes an optical density reference area, the optical density reference area including a material having a predetermined known optical density to calibrate the slide with an instrument being used to study the specimen cells.

Claim 21

Dependent Claim 35 words

A kit as recited in claim 12 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 22

Dependent Claim 35 words

A kit as recited in claim 14 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 23

Dependent Claim 35 words

A kit as recited in claim 15 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 24

Dependent Claim 35 words

A kit as recited in claim 1 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 25

Dependent Claim 35 words

A kit as recited in claim 4 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 26

Dependent Claim 35 words

A kit as recited in claim 9 wherein the reference means includes a reference material which is selected from the group consisting of trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 27

Dependent Claim 44 words

A kit as recited in claim 1 wherein the stain is selected from the group consisting essentially of thionin, Azure A, Azure C, pararosanilin, acriflavine, diaminobenzidine, alkaline phosphatase, eosin, methyl green, ethyl green, fast green, methylene blue, hematoxylin, acridine orange, giemsa and mixtures thereof.

Claim 28

Dependent Claim 184 words

A kit for the quantitation of nuclear DNA by light microscopy, the kit comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference area containing a reference means for staining, the reference means having physical characteristics which permit the reference means to associate with a stain proportionally to an association of the stain with DNA; one or more containers of stain material, the stain material including a stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers selected from the group consisting of thionin, Azure A, Azure C, pararosanilin, acriflavine and mixtures thereof in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 29

Dependent Claim 184 words

A kit for the quantitation of nuclear DNA by light microscopy, the kit comprising: a microscopic slide, the slide including a reference area and a specimen cell object area for receipt of specimen cell objects, the reference area containing a reference means for staining, the reference means having physical characteristics which permit the reference means to associate with a stain proportionally to an association of the stain with DNA; one or more containers of stain material, the stain material including a stain for simultaneous stain application to the specimen cell objects and the reference means, the stain in the containers selected from the group consisting of thionin, Azure A, Azure C, pararosanilin, acriflavine and mixtures thereof in an effective amount to provide an optical density to the reference means after staining for a predetermined time such that the optical density of the reference means will be substantially a linear function of stain concentration of the reference means after staining to determine a relationship between the stained reference means and the specimen cell objects to calibrate the instrument being used to study the specimen cell.

Claim 30

Dependent Claim 38 words

A kit as recited in claim 25 wherein the reference means includes a reference material which is selected from the group consisting of rat liver nuclei, trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 31

Dependent Claim 38 words

A kit as recited in claim 26 wherein the reference means includes a reference material which is selected from the group consisting of rat liver nuclei, trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 32

Dependent Claim 31 words

A kit for the quantitation of cell nuclei as recited in claim 1 wherein the stain is thionin and the kit is for the quantitation of nuclear DNA by light microscopy.

Claim 33

Dependent Claim 28 words

A kit as recited in claim 29 wherein the optical density of the reference means after staining is in the range of from about 0.1 to about 0.8.

Claim 34

Dependent Claim 38 words

A kit as recited in claim 30 wherein the reference means includes a reference material which is selected from the group consisting of rat liver nuclei, trout erythrocytes, chicken erythrocytes, dried DNA, cultured cell lines and mixtures thereof.

Claim 35

Dependent Claim 34 words

A kit for the quantitation of cell nuclei as recited in claim 1 wherein the kit is for the quantitation of nuclear DNA by light microscopy and the reference means includes rat liver nuclei.

Claim 36

Dependent Claim 49 words

A kit for the quantitation of cell nuclei as recited in claim 32 wherein the stain material includes thionin in an effective amount to provide an aqueous solution with a concentration of thionin in a range of from about 1 to about 5 mg/ml when mixed with 0.1N HCl.

Claim 37

Dependent Claim 46 words

A kit as recited in claim 32 wherein the microscopic slide further includes an optical density reference area, the optical density reference area including a material having a predetermined known optical density to calibrate the slide with an instrument being used to study the specimen cells.

Claim 38

Dependent Claim 28 words

A kit as recited in claim 33 wherein the optical density of the reference means after staining is in the range of from about 0.1 to about 0.8.

Claim 39

Dependent Claim 28 words

A kit as recited in claim 12 wherein the optical density of the reference means after staining is in the range of from about 0.1 to about 0.8.

Claim 40

Dependent Claim 28 words

A kit as recited in claim 26 wherein the optical density of the reference means after staining is in the range of from about 0.1 to about 0.8.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1995-04-28	1995-10-28	1996-04-28	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1999-04-28	1999-10-28	2000-04-28	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2003-04-28	2003-10-28	2004-04-28	$7700.00	Window Closed / Lapsed

Apparatus and method for analyses of biological specimens

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