Method of treating collected mammal semen and separating sperm into X Y components

• US 4,362,246 A
• Filed: 07/14/1980
• Issued: 12/07/1982
• Est. Priority Date: 07/14/1980
• Status: Expired due to Term

• Current Assignee: BTG International Inc. (Boston Scientific Corporation)
• Original Assignee: Edwin L. Adair
• Inventors: Adair, Edwin L.
• Primary Examiner(s): Knowles, Allen N.
• Application Number:
  US06/168,078
• Time in Patent Office: 876 Days
• Field of Search: 209/3.1, 209/3.2, 209/576, 209/578, 209/606, 209/3, 209/11, 23/230 B, 204/299, 204/180 R
• US Class Current: 209/3.3
• CPC Class Codes: A61K 35/52 Sperm; Prostate; Seminal fluid; Leydig cells of testes

Abstract

A method of treating mammal semen for use in artificial insemination is provided. Human saliva is added to the semen as an anti-coagulant. For mammals other than primates and voles, sperm-free human semen is added to the mammal semen as a cell membrane diffusion material so that a dye such as quinicrine or quinicrine mustard will be absorbed by the Y chromosome. The semen is then fed in a narrow stream through a detection chamber where it is irradiated with ultraviolet light causing the Y chromosomes to fluoresce. The stream is fed from the detection chamber through a fluidic amplifier wherein the stream is switched in response to whether the detected chromosome is fluorescent or non-fluorescent so that the fluorescent Y chromosome is directed through one outlet port and collected and the non-fluorescent X chromosome is directed through a second outlet port and collected.

Claims

1. A method of preserving semen for use in artificial insemination comprising the steps of:

collecting semen from a mammal;

treating the mammal semen by adding mammal saliva to prevent coagulation;

diluting the semen with a diluting solution; and

storing the semen in a water bath at substantially the body temperature of the mammal.

2. The method, as claimed in claim 1, wherein:

human saliva is added to the mammal semen to prevent coagulation.

3. The method, as claimed in claim 1, wherein:

a quantity of about 1.0 ml of human saliva is added to a quantity of about 2.0 ml to about 3.5 ml of mammal semen.

4. The method, as claimed in claim 1, wherein:

the mammal semen is diluted on about a 1;

1 ratio.

5. The method, as claimed in claim 1, wherein:

the treated and diluted mammal semen is stored at a temperature in the range of about 35°

C. to about 38.5°

C.

6. The method, as claimed in claim 4, wherein:

the treated and diluted semen is stored at a temperature of about 37°

C.

7. A method of separating X and Y chromosomes in mammal semen comprising the steps of:

collecting semen from the mammal;

adding about 1.0 ml of mammal saliva to about 2.0 ml to about 3.5 ml of mammal semen to prevent coagulation;

diluting the semen on a 1;

1 ratio with a diluting solution;

maintaining the temperature of the treated and diluted semen at a temperature of about 37°

C.;

adding a cell membrane diffusion material to the semen so that a dye can penetrate the cell membrane of the sperm containing the Y chromosome;

adding a dye to the semen to cause only the Y chromosome to fluoresce under ultraviolet light;

irradiating the semen with ultraviolet light;

moving the semen in a stream through a detection chamber;

sensing the presence of either a fluorescent Y chromosome or a non-fluorescent X chromosome in the detection chamber;

providing an output signal in response to the sensed chromosome which varies according to whether the chromosome is fluorescent or non-fluorescent;

passing the stream from the detection chamber through a fluidic amplifier; and

switching the stream in response to each sensed chromosome to one of two outlet ports so that the fluorescent Y chromosomes are directed through one outlet port and collected and the non-fluorescent X chromosomes are directed through the other outlet port and collected.

8. The method, as claimed in claim 7, wherein:

about 0.05 ml of DMSO is added to about 2.0 ml to about 3.5 ml of semen as the cell membrane diffusion material.

9. The method, as claimed in claim 7, wherein:

aboout 0.5 ml of sperm-free human semen is added to about 2.0 ml to about 3.5 ml of semen as the cell membrane diffusion material.

10. The method, as claimed in claim 7, wherein:

the mammal saliva is human saliva which is added to the semen.

11. The method, as claimed in claim 7, wherein:

the dye is from the group consisting of quinicrine, quinicrine hydrochloride, quinicrine dihydrochloride and quinicrine mustard.

12. A method of separating sperm containing X and Y chromosomes from semen which has been dyed to cause penetration of the dye only into the Y chromosome, comprising the steps of:

moving the semen in a stream through a detection chamber;

irradiating the semen with ultraviolet light;

sensing the presence of either a fluorescent or non-fluorescent chromosome in the stream in the detection chamber;

providing an output signal in response to the sensed chromosomes which varies according to whether the chromosome is fluorescent or non-fluorescent;

passing the stream from the detection chamber through a fluidic amplifier; and

switching the stream in response to each sensed chromosome in the detection chamber to direct the sensed chromosome to one of two outlet ports so that the fluorescent Y chromosomes are directed through one outlet port and collected and the non-fluorescent X chromosomes are directed through the other outlet port and collected.

13. The method, as claimed in claim 12, wherein:

said semen is irradiated with ultraviolet light having a wavelength between about 457 nm and about 488 nm.