Integration of biochemical protocols in a continuous flow microfluidic device

Abstract

Provided is a microfluidic device comprising a microfluidic substrate comprising at least one pathway for sample flow; and at least one thermal transfer member which is capable of cycling between at least two temperatures. The thermal transfer member is adapted to heat at least a portion of the sample pathway while a sample is flowing along said at least a portion of said sample pathway. Provided also are methods of carrying out biochemical protocols using such a device.

1 . A device comprising at least one inlet basin, at least one outlet basin and a microfluidic substrate comprising at least one sample pathway for sample flow; 
 said microfluidic substrate comprising at least one temperature regulated zone that cycles between at least two different and predetermined temperatures, said at least one temperature regulated zone being adapted to bring at least a portion of said sample pathway to said at least two temperatures while a sample is unidirectionally flowing along said at least a portion of said sample pathway and wherein a sample is cycled between said at least two different and predetermined temperatures while in said at least one temperature regulated zone.    
     
     
         2 . The device of  claim 1 , further comprising a force supplying member operably linked to said at least one pathway for sample flow wherein said force supplying member applies a force to said sample such that said sample travels along said at least one pathway.  
     
     
         3 . The device of  claim 2 , further comprising a sample supplier which supplies a sample to said at least one pathway.  
     
     
         4 . The device of  claim 3 , further comprising at least one inlet basin positioned at a first end of said at least one pathway such that said sample supplier supplies said sample to said inlet basin and said sample travels from said inlet basin to said at least one pathway.  
     
     
         5 . The device of  claim 4 , further comprising at least one outlet basin positioned at a second end of said pathway.  
     
     
         6 . The device of  claim 5 , further comprising at least one reagent supplier positioned between said inlet basin and said outlet basin.  
     
     
         7 . The device of  claim 6 , wherein said device comprises a plurality of said pathways.  
     
     
         8 . The device of  claim 7 , wherein said pathways comprise channels arranged in parallel.  
     
     
         9 . The device of  claim 8 , wherein the force generated by said force supplying member is pressure.  
     
     
         10 . The device of  claim 1 , wherein said microfluidic substrate consists essentially of silicon.  
     
     
         11 . The device of  claim 1 , further comprising a detector for measuring a physicochemical property of a biological sample.  
     
     
         12 . The device of  claim 1 , wherein said device further comprises temperature sensors in each temperature regulated zone.  
     
     
         13 . The device of  claim 12 , wherein said temperature sensors are two thermocouples and a platinum sensor.  
     
     
         14 . The device of  claim 1 , further comprising at least one sample that is continuously and unidirectionally flowing between said at least one inlet basin and said at least one outlet basin through said sample pathway.  
     
     
         15 . The device of  claim 1 , further comprising a series of samples that are continuously and unidirectionally flowing through said at least one sample pathway, said samples being separated by separators.  
     
     
         16 . A method for conducting a biochemical or chemical process comprising: 
 a) providing a device as set forth in  claim 1;     b) introducing at least one sample on said at least one sample pathway;    c) cycling at least a portion of at least one sample flow pathway between at least two temperatures while a sample comprising the reagents for said biochemical or chemical process is flowing through said at least a portion of said at least one sample flow pathway.    
     
     
         17 . The method of  claim 16 , wherein said sample flow pathway is located on a microfluidic substrate.  
     
     
         18 . The method of  claim 17 , wherein said sample flow pathway is in thermal communication with at least one thermal transfer member which cycles between said at least two temperatures while said sample is continuously flowing through said at least a portion of said at least one sample flow pathway.  
     
     
         19 . The method of  claim 18 , wherein said thermal transfer member cycles through said at least two temperatures a plurality of times while said sample is continuously flowing through said at least a portion of said at least one sample flow pathway.  
     
     
         20 . The method of  claim 19 , wherein said thermal transfer member cycles through said at least two temperatures from about 2 to about 35 times while said sample is continuously flowing through said at least a portion of said at least one sample flow pathway.  
     
     
         21 . The method of  claim 19 , wherein at a portion of a plurality of sample flow pathways are simultaneously cycled between said at least two temperatures while a plurality of samples are simultaneously flowing through said sample flow pathways.  
     
     
         22 . The method of  claim 21 , wherein said biochemical or chemical reaction comprises a nucleic acid amplification procedure.  
     
     
         23 . The method of  claim 22 , wherein said nucleic acid amplification procedure comprises polymerase chain reaction.  
     
     
         24 . The method of  claim 22 , further comprising determining the identity of at least one polymorphic nucleotide in the product of said nucleic acid amplification procedure.