Method and apparatuses for sorting objects in forensic DNA analysis and medical diagnostics

Abstract

The present invention relates to an apparatus and method of sorting objects and identifying the objects in a forensics sample, including using holographic optical trapping to sort objects from contaminants, and performing (single cell) PCR-based STR analysis on the objects to determine their identification. In addition, the chip used as a support for sorting the objects can also be used for performing single cell PCR-based STR analysis. In another embodiment, a microfluidics chip is used to stream the sample and sort the objects, before single cell PCR-based STR analysis is performed. The chip used for sorting utilizing HOT in the absence or presence of microfluidic streaming and sorting can also be the same as that used for the single cell PCR-based STR analysis.

1 . An apparatus for sorting objects, comprising:
 a holographic optical trapping apparatus, said optical trapping apparatus which optically traps and sorts a sample of wanted objects from unwanted objects;   a microfluidics chip containing an input chamber in which said sample is disposed, for streaming said sample from said input chamber to a plurality of output chambers;   wherein said sorting of said sample into said wanted objects and said unwanted objects takes place at said input chamber using said holographic optical trapping apparatus;   wherein said sorting removes said wanted objects individually into each of said plurality of output chambers, such that each of said output chambers contains a single wanted object; and   an apparatus which performs single-cell PCR-based STR analysis on each of said single wanted objects, said apparatus being disposed downstream from said plurality of output chambers.   
     
     
         2 . The apparatus of  claim 1 , wherein said wanted objects are cells and said unwanted objects are contaminants. 
     
     
         3 . The apparatus of  claim 1 , wherein said wanted objects are normal cells and said unwanted objects are cancerous cells or vice versa. 
     
     
         4 . The apparatus of  claim 1 , wherein said sorting and said PCR-based STR analysis are performed on a single microfluidics chip. 
     
     
         5 . The apparatus of  claim 1 , further comprising:
 a plurality of valves disposed at said input chamber, to prevent backflow.   
     
     
         6 . The apparatus according to  claim 1 , further comprising:
 a filter disposed at an outlet of said inlet chamber.   
     
     
         7 . The apparatus according to  claim 1 , wherein said sorting and PCR-based STR analysis are automated and multiplexed. 
     
     
         8 . An apparatus for sorting objects, comprising:
 a microfluidics chip containing an input chamber and a plurality of output chambers, said input chamber containing a sample of wanted objects and unwanted objects to be separated;   wherein said microfluidics chip streams said sample from said input chamber into said plurality of output chambers, such that said wanted objects are sorted from said unwanted objects;   wherein said sorting removes said wanted objects individually into each of said plurality of output chambers. such that each of said output chambers contains a single wanted object: and   an apparatus which performs single-cell PCR-based STR analysis on each of said single wanted objects, said apparatus being disposed downstream from said plurality.   
     
     
         9 . The apparatus of  claim 8 , further comprising:
 a holographic optical trapping apparatus which sorts said wanted objects from said unwanted objects at said input chamber, and removes said wanted objects into said plurality of output chambers.   
     
     
         10 . The apparatus of  claim 8 , wherein said microfluidics chip and said apparatus for performing said PCR-based STR analysis are provided on a single chip. 
     
     
         11 . The apparatus of  claim 8 , wherein said objects are cells, and said unwanted objects are contaminants. 
     
     
         12 . The apparatus of  claim 8 , wherein said wanted objects are normal cells and said unwanted objects are cancerous cells. 
     
     
         13 . The apparatus of  claim 8 , further comprising:
 a plurality of valves disposed at said input chamber, to prevent backflow.   
     
     
         14 . The apparatus according to  claim 8 , further comprising:
 a filter disposed at an outlet of said inlet chamber.   
     
     
         15 . The apparatus according to  claim 8 , wherein said sorting and PCR-based STR analysis are automated and multiplexed. 
     
     
         16 . The apparatus according to  claim 1 , wherein said wanted objects are sperm cells. 
     
     
         17 . The apparatus according to  claim 8 , wherein said wanted objects are sperm Cells. 
     
     
         18 . The apparatus according to  claim 1 , wherein streaming of said sample is controlled such that no two objects from said sample occupy a same position in a channel or at a junction in said microfluidics chip. 
     
     
         19 . The apparatus according to  claim 1 , wherein said unwanted objects settle in said input chamber, and said wanted objects flow to said output chamber. 
     
     
         20 . The apparatus according to  claim 1 , wherein each of said single wanted objects flows from each of said individual output chambers to a corresponding individual lysis chamber where said PCR-based STR analysis is performed on each of said single wanted objects. 
     
     
         21 . The apparatus according to  claim 1 , wherein said wanted objects are streamed into a single output chamber prior to being streamed into said individual output chambers. 
     
     
         22 . The apparatus according to  claim 8 , wherein streaming of said sample is controlled such that no two objects from said sample occupy a same position in a channel or at a junction in said microfluidics chip. 
     
     
         23 . The apparatus according to  claim 8 , wherein said unwanted objects settle in said input chamber, and said wanted objects flow into said plurality of output chambers. 
     
     
         24 . The apparatus according to  claim 8 , wherein each of said single wanted objects flows from each of said individual output chambers to a corresponding individual lysis chamber where said PCR-based STR analysis is performed on each of said single wanted objects. 
     
     
         25 . The apparatus according to  claim 8 , wherein said wanted objects are streamed into a single output chamber prior to being streamed into said individual output chambers. 
     
     
         26 . The apparatus according to  claim 1 , wherein said microfluidics chip is optically transparent. 
     
     
         27 . The apparatus according to  claim 9 , wherein said microfluidics chip isoptically transparent.