US3586859A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Fluorescent cell viability counter
Assignee: Individual
•
Filing: 1970-02-16
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Granted: 1971-06-22
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Expires: 1990-02-16
Abstract
Abstract Method and apparatus for automatically ascertaining the number of dead cells in a batch. All of the cells in the batch are stained with a specific fluorescent dye in a reservoir, then the cells are passed through a capillary tube by means of a peristaltic pump under a microscope while illuminated by an ultraviolet light. The microscope is coupled to a photomultiplier which in turn has its output applied to an electronic counter with a readout. After the dead cells are counted, the batch is immersed in a heating bath in which all the viable cells are killed and another count is obtained by repassing the batch beneath the microscope. The difference in the counts is the number of original viable cells while the first count is the original number of dead cells.
Inventors
- 👤 Irwin J Katz
- 👤 Nicholas M Satriano
- 👤 Morris A Benjaminson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 19:12
CPC Classifications
G; G06; G06M; G06M1/00; G06M1/08; G06M1/10; G06M1/101; C; C12; C12M; C12M41/00; C12M41/30; C12M41/36; G01; G01N; G01N15/00; G01N15/10; G01N15/14; G01N2015/1486; Y; Y10; Y10S; Y10S435/00; Y10S435/808
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
181 words
An apparatus for determining the number of viable and nonviable cells in a 5mg./ml. saline solution of 4 acetamido 440 isothiocyanostilbene 2,2 disulfonic acid Disodium which comprises: an elongated restricted transparent passageway having an open end and connected at its other end to a circulating means for flowing a liquid through said passageway, the opposite end of said circulating means being connected to means for killing said viable cells; coupling means connecting said killing means to said open end of said passageway; a fluorescent microscope having an objective disposed on one side of said passageway; a source of ultraviolet light disposed on the opposite side of said passageway from said objective for directing ultraviolet light through said passageway into said objective; light amplifying and detecting means connected to said microscope for detecting light pulses therefrom and converting them to electric signals and, computer means connected to said detecting means for counting said light pulses; and whereby when the solution is circulated twice through said apparatus a count oF the number of nonviable and the total number of cells will be obtained.
Claim 2
Independent Claim
181 words
An apparatus for determining the number of viable and nonviable cells in a 5mg./ml. saline solution of 4 acetamido 440 isothiocyanostilbene 2,2 disulfonic acid Disodium which comprises: an elongated restricted transparent passageway having an open end and connected at its other end to a circulating means for flowing a liquid through said passageway, the opposite end of said circulating means being connected to means for killing said viable cells; coupling means connecting said killing means to said open end of said passageway; a fluorescent microscope having an objective disposed on one side of said passageway; a source of ultraviolet light disposed on the opposite side of said passageway from said objective for directing ultraviolet light through said passageway into said objective; light amplifying and detecting means connected to said microscope for detecting light pulses therefrom and converting them to electric signals and, computer means connected to said detecting means for counting said light pulses; and whereby when the solution is circulated twice through said apparatus a count oF the number of nonviable and the total number of cells will be obtained.
Claim 3
Dependent Claim
14 words
The apparatus according to claim 1 wherein said restricted passageway is a capillary tube.
Claim 4
Dependent Claim
14 words
The apparatus according to claim 2 wherein said circulating means is a peristaltic pump.
Claim 5
Dependent Claim
21 words
The apparatus according to claim 3 wherein said means for killing is a heating module through which the cells are passed.
Claim 6
Dependent Claim
18 words
The apparatus according to claim 4 wherein said amplifying and detecting means is a photomultiplier circuit and tube.
Claim 7
Dependent Claim
23 words
The apparatus according to claim 5 further including a specimen reservoir disposed in line with and proximate said open end of said passageway.
Claim 8
Independent Claim
89 words
A method for ascertaining the number of viable and nonviable cells within a specimen which comprises the steps of: preparing a solution of said specimen cells with a saline solution of a nontoxic fluorescent dye; passing said prepared solution through a restricted passage; exposing said prepared solution in said passage to an ultraviolet light; detecting and counting those cells which fluoresce; killing any viable cells in said prepared solution; and repassing said prepared solution after killing said viable cells through said passage and detecting and counting the fluorescent cells.
Claim 9
Independent Claim
89 words
A method for ascertaining the number of viable and nonviable cells within a specimen which comprises the steps of: preparing a solution of said specimen cells with a saline solution of a nontoxic fluorescent dye; passing said prepared solution through a restricted passage; exposing said prepared solution in said passage to an ultraviolet light; detecting and counting those cells which fluoresce; killing any viable cells in said prepared solution; and repassing said prepared solution after killing said viable cells through said passage and detecting and counting the fluorescent cells.
Claim 10
Dependent Claim
19 words
The method according to claim 7 wherein said dye is 4 acetamido 4'' isothiocyanostilbene 2, 2 disulfonic acid Disodium.
Claim 11
Dependent Claim
28 words
The method according to claim 8 wherein said prepared solution consists of said specimen cells added to 5 mg. of said dye for every ml. of saline solution.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1974-06-21 | 1974-12-21 | 1975-06-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1978-06-21 | 1978-12-21 | 1979-06-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1982-06-21 | 1982-12-21 | 1983-06-22 | $7700.00 | Window Closed / Lapsed |