Patent US4412004A successfully refreshed from Google Patents.
US4412004A
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Method for treating red blood cells to effect sphering and reagent therefor
Assignee: Bayer Corp
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Filing: 1981-06-26
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Granted: 1983-10-25
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Expires: 2001-06-26
Abstract
Abstract A method is disclosed of blood serum sample preparation for improved, more accurate and precise, electro-optical method for measuring erythrocyte volumes, individually and as an average.
Inventors
- 👤 Leonard Ornstein
- 👤 Young R. Kim
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 03:07
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/96; G01N2496/00; G01N2496/05; Y; Y10; Y10T; Y10T436/00; Y10T436/10; Y10T436/101666; Y10T436/106664
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
182 words
A diagnostic method for treating mammalian red blood cells in a sample to provide a sample which can be effectively measured electrooptically for determination of red blood cell volumes which process comprises combining an anticoagulated whole blood sample with a first isotonic solution containing sphering agent, and treating an aliquot of the resulting sample with a second isotonic solution containing a protein which reversibly binds the sphering agent and containing a sphering agent wherein the weight ratio of protein sphering agent in the aliquot and the final sample is from about 20:1 to about 70:1 and the concentration of sphering agent in said final sample is from about 2 mg./100 ml. to about 10 mg./100 ml.; and whereby the presence of the prescribed concentration of sphering agent could lead to lysis if sufficient of said protein were not present and a low red blood cell count existed in the original sample; and whereby the presence of less than the prescribed concentration of the sphering agent could lead to incomplete sphering if a high red blood cell count existed in the original sample.
Claim 2
Dependent Claim
182 words
A diagnostic method for treating mammalian red blood cells in a sample to provide a sample which can be effectively measured electrooptically for determination of red blood cell volumes which process comprises combining an anticoagulated whole blood sample with a first isotonic solution containing sphering agent, and treating an aliquot of the resulting sample with a second isotonic solution containing a protein which reversibly binds the sphering agent and containing a sphering agent wherein the weight ratio of protein sphering agent in the aliquot and the final sample is from about 20:1 to about 70:1 and the concentration of sphering agent in said final sample is from about 2 mg./100 ml. to about 10 mg./100 ml.; and whereby the presence of the prescribed concentration of sphering agent could lead to lysis if sufficient of said protein were not present and a low red blood cell count existed in the original sample; and whereby the presence of less than the prescribed concentration of the sphering agent could lead to incomplete sphering if a high red blood cell count existed in the original sample.
Claim 3
Dependent Claim
26 words
The method of claim 1 wherein said whole blood sample is prediluted with saline as diluent resulting in about a 50% by volume dilution of sample.
Claim 4
Dependent Claim
24 words
The method of claim 2 wherein said first isotonic solution combining step results in a dilution of original sample of about 1:50 by volume.
Claim 5
Dependent Claim
25 words
The method of claim 2 wherein said second isotonic solution treatment step results in a further dilution of original sample of about 1:1000 by volume.
Claim 6
Dependent Claim
32 words
The method of claim 2 wherein the sphering agent employed in the dilution steps is an alkali metal salt of an alkyl sulfate, said alkyl containing from 10 to 16 carbon atoms.
Claim 7
Dependent Claim
16 words
The method of claim 5 wherein said alkyl sulfate salt is an alkali metal lauryl sulfate.
Claim 8
Dependent Claim
15 words
The method of claim 6 wherein said alkali metal lauryl sulfate is sodium lauryl sulfate.
Claim 9
Dependent Claim
19 words
The method of claim 2 wherein the protein employed in said isotonic solution treatment step is a serum albumin.
Claim 10
Dependent Claim
31 words
The method of claim 2 wherein, in the final sample, the weight ratio of protein/sphering agent is about 50:1 and the total concentration of sphering agent is about 3 mg./100 ml.
Claim 11
Dependent Claim
32 words
The method of claim 2 wherein the final sample which contains sphered red blood cells without volume change is subjected to light scattering measurement for red blood cell count and volume determination.
Claim 12
Dependent Claim
11 words
The method of claim 2 effected in a continuous, automated manner.
Claim 13
Dependent Claim
12 words
The method of claim 2 effected in a discontinuous or discrete manner.
Claim 14
Dependent Claim
66 words
The method of claim 1 wherein instead of said second isotonic solution treatment step, said aliquot sample is treated with an isotonic solution containing fixing agent, and the aliquot resulting after the first dilution step is characterized by an endogenous protein/sphering agent weight ratio of from about 20:1 to about 70:1 and sphering agent concentration of from about 2 mg./100 ml. to about 10 mg./100 ml.
Claim 15
Dependent Claim
28 words
The method of claim 13 wherein said fixing agent solution contains glutaraldehyde in an amount to provide a final glutaraldehyde concentration of from 0.1% to 0.4% by weight.
Claim 16
Dependent Claim
20 words
The method of claim 13 wherein said fixing agent solution contains saline or a saline-sphering agent mixture adjusted to isotonicity.
Claim 17
Dependent Claim
12 words
The method of claim 15 wherein a saline-sphering agent mixture is employed.
Claim 18
Dependent Claim
26 words
The method of claim 13 wherein said whole blood sample is prediluted with saline as diluent resulting in about a 50% by volume dilution of sample.
Claim 19
Dependent Claim
25 words
The method of claim 17 wherein said first isotonic solution combining step results in a further dilution of original sample to about 1:50 by volume.
Claim 20
Dependent Claim
22 words
The method of claim 17 wherein said fixing step results in a further dilution of original sample to about 1:1000 by volume.
Claim 21
Dependent Claim
32 words
The method of claim 17 wherein the sphering agent employed in the dilution steps is an alkali metal salt of an alkyl sulfate, said alkyl containing from 10 to 16 carbon atoms.
Claim 22
Dependent Claim
16 words
The method of claim 20 wherein said alkyl sulfate salt is an alkali metal lauryl sulfate.
Claim 23
Dependent Claim
15 words
The method of claim 21 wherein said alkali metal lauryl sulfate is sodium lauryl sulfate.
Claim 24
Dependent Claim
12 words
The method of claim 17 wherein said endogenous protein is plasma protein.
Claim 25
Dependent Claim
33 words
The method of claim 17 wherein the weight ratio of endogenous protein/sphering agent before addition of fixing agent is about 50:1 and the total concentration of sphering agent is about 3 mg./100 ml.
Claim 26
Dependent Claim
30 words
The method of claim 17 wherein the final sample which contains fixed, sphered red blood cells is subjected to light scattering measurement for red blood cell count and volume determination.
Claim 27
Dependent Claim
11 words
The method of claim 17 effected in a continuous, automated manner.
Claim 28
Dependent Claim
12 words
The method of claim 17 effected in a discontinuous or discrete manner.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-10-24 | 1987-04-25 | 1987-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-10-24 | 1991-04-25 | 1991-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-10-24 | 1995-04-25 | 1995-10-25 | $7700.00 | Window Closed / Lapsed |