Microfluidic system and method with focused energy apparatus

Abstract

A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.

What is claimed is: 
     
         1 . A method of producing a subpopulation of sperm enriched for a desired sex chromosome comprising:
 a) generating a stream containing a population of sperm;   b) flowing the stream containing the population of sperm cells through a channel;   c) focusing, positioning, and orienting the population of sperm cells in the channel without a nozzle;   d) differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells;   e) acting on the differentiated population of sperm to form the subpopulation of sperm enriched for the desired sex chromosome and a subpopulation of sperm enriched for an undesired sex chromosome; and   f) collecting the subpopulation of sperm enriched for the desired sex chromosome and the subpopulation of sperm enriched for the undesired sex chromosome together.   
     
     
         2 . The method of  claim 1 , wherein differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells comprises:
 a) interrogating the population of sperm with a source of electromagnetic radiation;   b) detecting signals produced in response to interrogation with the source of electromagnetic radiation; and   c) identifying sperm in the population of sperm that do not have the desired sex chromosome based on the detected signals.   
     
     
         3 . The method of  claim 2 , wherein the source of electromagnetic radiation is an LED light source. 
     
     
         4 . The method of  claim 2 , wherein the source of electromagnetic radiation is a detection laser. 
     
     
         5 . The method of  claim 2 , wherein the electromagnetic radiation is emitted as a continuous wave or in pulses. 
     
     
         6 . The method of  claim 2 , wherein the signals produced in response to interrogation with the source of electromagnetic radiation comprise how sperm cells in the population of sperm reflect, refract, diffract, absorb, and/or emit the electromagnetic radiation. 
     
     
         7 . The method of  claim 1 , wherein acting on the differentiated population of sperm to form the subpopulation of sperm enriched for the desired sex chromosome and a subpopulation of sperm enriched for an undesired sex chromosome comprises ablating an undesired subpopulation at a predetermined delay time from selecting the desired subpopulation. 
     
     
         8 . The method of  claim 1 , wherein the step of collecting comprises collecting the subpopulation in a collector. 
     
     
         9 . A method of producing at least one fertilized egg from a female mammal, the method comprising:
 a) obtaining a population of sperm from a raw ejaculate sample;   b) generating a stream containing the population of sperm;   c) flowing the stream containing the population of sperm cells through a channel;   d) focusing, positioning, and orienting the population of sperm cells in the channel without a nozzle;   e) differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells;   f) selecting a desired subpopulation;   g) acting on the identified sperm from the population of sperm to form the subpopulation of sperm enriched for the desired sex chromosome;   h) collecting the subpopulation of sperm enriched for the desired sex chromosome and the subpopulation of sperm enriched for the undesired sex chromosome together; and   i) mixing at least a portion of the collected sperm cells with at least one egg from a female mammal to produce a fertilized egg.   
     
     
         10 . The method of  claim 9 , wherein differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells comprises:
 a) interrogating the population of sperm with a source of electromagnetic radiation;   b) detecting signals produced in response to interrogation with the source of electromagnetic radiation; and   c) identifying sperm in the population of sperm that do not have the desired sex chromosome based on the detected signals.   
     
     
         11 . The method of  claim 10 , wherein the source of electromagnetic radiation is an LED light source. 
     
     
         12 . The method of  claim 10 , wherein the source of electromagnetic radiation is a detection laser. 
     
     
         13 . The method of  claim 10 , wherein the electromagnetic radiation is emitted as a continuous wave or in pulses. 
     
     
         14 . The method of  claim 10 , wherein the signals produced in response to interrogation with the source of electromagnetic radiation comprise how sperm cells in the population of sperm reflect, refract, diffract, absorb, and/or emit the electromagnetic radiation. 
     
     
         15 . The method of  claim 9 , wherein acting on the differentiated population of sperm to form the subpopulation of sperm enriched for the desired sex chromosome and a subpopulation of sperm enriched for an undesired sex chromosome comprises ablating an undesired subpopulation at a predetermined delay time from selecting the desired subpopulation. 
     
     
         16 . The method of  claim 9 , wherein the step of collecting comprises collecting the subpopulation in a collector. 
     
     
         17 . The method of  claim 9  further comprising staining the population of sperm with a dye. 
     
     
         18 . The method of  claim 9  further comprising inserting at least a portion of the collected sperm cells into a female mammal, wherein fertilizing the at least one egg from said female mammal produces at least one embryo from said female mammal. 
     
     
         19 . A method of producing a subpopulation of sperm enriched for a desired sex chromosome comprising:
 a) generating a stream containing a population of sperm cells in a channel;   b) focusing, positioning, and orienting the population of sperm cells in the channel without a nozzle;   c) interrogating the population of sperm cells to identify a desired subpopulation of sperm cells and an undesired subpopulation of sperm cells;   d) acting on the undesired subpopulation of sperm cells; and   e) collecting the population of sperm cells.   
     
     
         20 . The method of  claim 19 , wherein acting on the undesired subpopulation of sperm cells comprises ablating the undesired subpopulation of sperm cells.