US5403720A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method for rapid measurement of living microorganisms and measuring kit
Assignee: Idemitsu Kosan Co Ltd
•
Filing: 1993-12-06
•
Granted: 1995-04-04
•
Expires: 2013-12-06
Abstract
Abstract A method for measuring the number of living microorganisms in a specimen which comprises entrapping the microorganisms on a hydrophobic filter after the dyeing thereof, or alternatively dyeing the microorganisms after entrapping them on the hydrophobic filter, removing excessive coloring matter by washing, and then determining the number of the microorganisms by the degree of coloration thereof. A kit for measuring the number of living microorganisms which comprises a hydrophobic filter, a syringe to which the filter is fittable, a coloring matter solution, a cleaning solution, and a color reference table. The present invention enables the rapid and convenient measurement of the number of living microorganisms in the specimen without use of special equipment. In accordance with the present invention, the measurement is usually completed within ten minutes.
Inventors
- 👤 Mikio Sato
- 👤 Futoshi Kawane
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 12:59
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/06; Y; Y10; Y10S; Y10S436/00; Y10S436/825; Y10S436/826; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
136 words
A method for measuring the number of living microorganisms selected from the group consisting of fungi, yeast and bacteria in a specimen solution which comprises dyeing the microorganisms selected from the group consisting of fungi, yeast and bacteria with a water-soluble coloring matter solution, the water-soluble coloring matter being in a concentration of 0.0005 to 2.0% and entrapping the dyed microorganisms on a hydrophobic filter made of polytetrafluoroethylene, or alternatively entrapping the microorganisms on said hydrophobic filter and dyeing the entrapped microorganisms with said coloring matter solution, removing excessive coloring matter solution by washing with a washing solution consisting essentially of (i) water or a buffer solution, having a pH of 6 to 8 and (ii) 0.0001 to 1% of a surfactant, and then determining the number of the microorganisms by the degree of coloration thereof.
Claim 2
Independent Claim
136 words
A method for measuring the number of living microorganisms selected from the group consisting of fungi, yeast and bacteria in a specimen solution which comprises dyeing the microorganisms selected from the group consisting of fungi, yeast and bacteria with a water-soluble coloring matter solution, the water-soluble coloring matter being in a concentration of 0.0005 to 2.0% and entrapping the dyed microorganisms on a hydrophobic filter made of polytetrafluoroethylene, or alternatively entrapping the microorganisms on said hydrophobic filter and dyeing the entrapped microorganisms with said coloring matter solution, removing excessive coloring matter solution by washing with a washing solution consisting essentially of (i) water or a buffer solution, having a pH of 6 to 8 and (ii) 0.0001 to 1% of a surfactant, and then determining the number of the microorganisms by the degree of coloration thereof.
Claim 3
Dependent Claim
46 words
The method as claimed in claim 1, wherein the coloring matter which is attached to the microorganisms is eluted with an organic solvent, and the degree of coloration of the elute is measured for optical density and determined by colorimetry with reference to a calibration curve.
Claim 4
Dependent Claim
15 words
The method as claimed in claim 1, wherein the specimen is water-soluble metal working fluid.
Claim 5
Dependent Claim
13 words
The method as claimed in claim 1, wherein the microorganisms are aerobic bacteria.
Claim 6
Dependent Claim
15 words
The method as claimed in claim 1, wherein the coloring matter is fuchsine or safranine.
Claim 7
Dependent Claim
88 words
A kit for measuring the number of living microorganisms selected from the group consisting of fungi, yeast and bacteria in a specimen solution which comprises a hydrophobic filter made of polytetrafluoroethylene, a syringe to which the filter is fittable, a water-soluble coloring matter solution, the water-soluble coloring matter being in a concentration of 0.0005 to 2.0%, a washing solution consisting essentially of (i) water or a buffer solution having a pH of 6 to 8 and (ii) 0.0001 to 1% of a surfactant, and a color reference table.
Claim 8
Dependent Claim
88 words
A kit for measuring the number of living microorganisms selected from the group consisting of fungi, yeast and bacteria in a specimen solution which comprises a hydrophobic filter made of polytetrafluoroethylene, a syringe to which the filter is fittable, a water-soluble coloring matter solution, the water-soluble coloring matter being in a concentration of 0.0005 to 2.0%, a washing solution consisting essentially of (i) water or a buffer solution having a pH of 6 to 8 and (ii) 0.0001 to 1% of a surfactant, and a color reference table.
Claim 9
Dependent Claim
20 words
The kit as claimed in claim 6, wherein the coloring matter solution is fuchsine dyeing solution or safranine dyeing solution.
Claim 10
Dependent Claim
18 words
The kit as claimed in claim 6, which is used for measuring the number of living aerobic bacteria.
Claim 11
Dependent Claim
19 words
The method as claimed in claim 1, wherein the entrapping and washing are carried out by filtration under pressure.
Claim 12
Dependent Claim
25 words
The method as claimed in claim 1, wherein the specimen is selected from the group consisting of a liquid foodstuff, a water-soluble paint and water.
Claim 13
Dependent Claim
28 words
The method as claimed in claim 2, wherein the ratio of the amount of the coloring matter solution to that of the specimen solution is at least 1:1.
Claim 14
Dependent Claim
40 words
The method as claimed in claim 11, wherein the concentration of the coloring matter solution is 0.002 to 1%; and the ratio of the amount of the coloring matter solution to that of the specimen solution is at least 5:1.
Claim 15
Dependent Claim
56 words
The method as claimed in claim 12, wherein the coloring matter solution is selected from the group consisting of a fuchsine dyeing solution, a safranine dyeing solution and a Victoria Blue dyeing solution; and the specimen is selected from the group consisting of a water-soluble metal working fluid, a liquid foodstuff, a water-soluble paint and water.
Claim 16
Dependent Claim
43 words
The method as claimed in claim 13, wherein the coloring matter solution is selected from the group consisting of a fuchsine dyeing solution and a safranine dyeing solution; the microorganisms are aerobic bacteria; and the entrapping and washing are carried out under pressure.
Claim 17
Dependent Claim
25 words
The method as claimed in claim 2, wherein the hydrophobic filter has a pore diameter of 0.22 to 0.50 mμ and the microorganisms are bacteria.
Claim 18
Dependent Claim
20 words
The method as claimed in claim 15, wherein the washing solution is in an amount of 1 to 5 ml.
Claim 19
Dependent Claim
28 words
The method as claimed in claim 16, wherein the specimen is a water-soluble working fluid; the coloring matter is fuchsine or safranine and the microorganisms are aerobic bacteria.
Claim 20
Dependent Claim
20 words
The kit as claimed in claim 7, wherein the hydrophobic filter has a pore diameter of 0.22 to 50 mμ.
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) | 1998-04-03 | 1998-10-03 | 1999-04-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-04-03 | 2002-10-03 | 2003-04-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-04-03 | 2006-10-03 | 2007-04-04 | $7700.00 | Window Closed / Lapsed |