Prediction of fertility in males

Abstract

A method for evaluating sperm fertility. The method includes the steps of obtaining a sample of sperm from an animal of a species; staining the sample with a fluorescent DNA-binding dye; collecting at least one image of the stained sample; determining an edge of a nucleus of at least one sperm within the stained sample in the at least one image; measuring an intensity of the DNA-binding dye within an area defined by the edge of the nucleus of the at least one sperm; determining an average intensity per unit area of the area defined by the edge of the nucleus of the at least one sperm; comparing the average intensity per unit area to an average intensity per unit area for high-fertility sperm and low-fertility sperm of the same species to determine if the sample has high or low fertility.

1 - 18 . (canceled) 
     
     
         19 . A method for evaluating sperm fertility, comprising the steps of:
 obtaining a sample of sperm from an animal;   staining the sample with a fluorescent DNA (deoxyribonucleic acid)-binding dye;   collecting at least one image of the stained sample;   determining an edge of a nucleus of at least one sperm within the stained sample in the at least one image;   measuring an intensity of the DNA-binding dye within an area defined by the edge of the nucleus of the at least one sperm;   determining an average intensity per unit area of the area defined by the edge of the nucleus of the at least one sperm; and   comparing the average intensity per unit area to an average intensity per unit area for a high-fertility sperm from an animal of the same species and to an average intensity per unit area for a low-fertility sperm from an animal of the same species to determine if the sample has high or low fertility.   
     
     
         20 . The method of  claim 19 , wherein staining the sample with a fluorescent DNA-binding dye comprises staining the sample with Hoechst 33342. 
     
     
         21 . The method of  claim 19 , wherein staining the sample with a fluorescent DNA-binding dye further comprises applying the stained sample to a substrate. 
     
     
         22 . The method of  claim 21 , wherein the nucleus of the at least one sperm within the stained sample comprises a flattened oval and wherein the flattened portion is adjacent to the substrate. 
     
     
         23 . The method of  claim 19 , wherein obtaining a sample of sperm from an animal comprises obtaining a sample of sperm from a bull, a boar, a human, a horse, a ram, or a dog. 
     
     
         24 . The method of  claim 19 , wherein collecting at least one image of the stained sample comprises deconvolving the at least one image to remove out of focus information. 
     
     
         25 . The method of  claim 19 , wherein the high-fertility sperm has a lower intensity per unit area than the low-fertility sperm. 
     
     
         26 . A method for evaluating sperm fertility, comprising the steps of:
 obtaining a sample of sperm from an animal;   staining the sample with a fluorescent DNA-binding dye;   obtaining fluorescent intensity measurements from a plurality of sperm in the stained sample;   determining an average intensity of the fluorescent intensity measurements obtained from the plurality of sperm in the stained sample; and   comparing the average intensity to an average intensity for a high-fertility sperm from an animal of the same species and to an average intensity for a low-fertility sperm from an animal of the same species.   
     
     
         27 . The method of  claim 26 , wherein obtaining fluorescent intensity measurements from a plurality of sperm in the stained sample further comprises collecting at least one image of the stained sample. 
     
     
         28 . The method of  claim 27 , wherein determining an average intensity of the fluorescent intensity measurements obtained from the plurality of sperm in the stained sample further comprises
 determining an edge of a nucleus of at least one sperm within the stained sample in the at least one image,   measuring an intensity of the DNA-binding dye within an area defined by the edge of the nucleus of the at least one sperm, and   determining an average intensity per unit area of the area defined by the edge of the nucleus of the at least one sperm.   
     
     
         29 . The method of  claim 28 , wherein collecting at least one image of the stained sample comprises deconvolving the at least one image to remove out of focus information. 
     
     
         30 . The method of  claim 29 , wherein staining the sample with a fluorescent DNA-binding dye further comprises applying the stained sample to a substrate. 
     
     
         31 . The method of  claim 30 , wherein the nucleus of the at least one sperm within the stained sample comprises a flattened oval and wherein the flattened portion is adjacent to the substrate. 
     
     
         32 . The method of  claim 26 , wherein obtaining fluorescent intensity measurements from a plurality of sperm in a sample comprises obtaining fluorescent intensity measurements using flow cytometry. 
     
     
         33 . The method of  claim 32 , wherein obtaining fluorescent intensity measurements using flow cytometry comprises obtaining fluorescent intensity measurements using flow cytometry with an orienting nozzle. 
     
     
         34 . The method of  claim 26 , wherein obtaining a sample of sperm from an animal comprises obtaining a sample of sperm from a bull, a boar, a human, a horse, a ram, or a dog. 
     
     
         35 . The method of  claim 26 , wherein staining the sample with a fluorescent DNA-binding dye comprises staining the sample with Hoechst 33342. 
     
     
         36 . The method of  claim 26 , wherein the high-fertility sperm has a lower average intensity than the low-fertility sperm. 
     
     
         37 . The method of  claim 19 , wherein collecting the at least one image of the stained sample is performed on a fluorescent microscope. 
     
     
         38 . The method of  claim 19 , wherein if the sample has an average intensity per unit area that is significantly higher than the average intensity per unit area for the high-fertility sperm from the animal of the same species as measured by having a p value of <0.05, the sample is identified as having low-fertility sperm.