Methods and systems for continuous flow cell lysis in a microfluidic device

Abstract

The present invention relates to methods and systems for cell lysis in a microfluidic device. More specifically, embodiments of the present invention relate to methods and systems for rapid continuous flow mechanical cell lysis. In one embodiment, a microfluidic device includes one or more microfluidic channels, each channel comprising constricted regions and non-constricted regions separating the constricted regions, wherein the constricted regions are configured to disrupt the cellular membranes of cells in fluid flowing through the one or more microfluidic channels.

What is claimed is: 
     
         1 . A microfluidic device for lysing cells, the microfluidic device comprising:
 one or more microfluidic channels, each channel comprising constricted regions and non-constricted regions separating the constricted regions, wherein the constricted regions are configured to disrupt the cellular membranes of cells in fluid flowing through the one or more microfluidic channels.   
     
     
         2 . The device of  claim 1 , wherein the one or more microfluidic channels are parallel to each other. 
     
     
         3 . The device of  claim 1 , wherein the non-constricted regions are arranged in a honeycomb-like pattern. 
     
     
         4 . The device of  claim 1 , wherein each of the non-constricted regions has a width of 40-100 μm, and each of the constricted regions has a width of 3-10 μm. 
     
     
         5 . The device of  claim 1 , wherein each of the non-constricted regions has a length of 60-120 μm, and each of the constricted regions has a length of 10-20 μm. 
     
     
         6 . The device of  claim 1 , wherein there are 1-40 microfluidic channels. 
     
     
         7 . The device of  claim 1 , wherein each microfluidic channel comprises 3-15 constricted regions. 
     
     
         8 . The device of  claim 1 , wherein each constricted region has a width of 3-10 μm. 
     
     
         9 . The device of  claim 1 , wherein each microfluidic channel comprises a first segment of constricted regions along a fluid flow path having a width of 6-8 μm and a second segment of constricted regions along a fluid flow path having a width of 4-6 μm. 
     
     
         10 . The device of  claim 1 , wherein each microfluidic channel comprises 10 constricted regions. 
     
     
         11 . The device of  claim 10 , wherein for each channel, the first five constricted regions along a fluid flow path have a width of 6.5 μm, and the last five constricted regions along the fluid flow path have a width of 5 μm. 
     
     
         12 . The device of  claim 1 , wherein each of the constricted regions has a width of 2.5-4.5 μm. 
     
     
         13 . The device of  claim 1 , wherein each of the microfluidic channels has 4 constricted regions and each constricted region has a width of 3 μm. 
     
     
         14 . The device of  claim 1 , wherein there are 20 microfluidic channels. 
     
     
         15 . The device of  claim 1 , wherein there are 40 microfluidic channels. 
     
     
         16 . The device of  claim 1 , wherein the microfluidic channels are configured to support a flow rate from about 20 μL/min to about 2000 μL/min. 
     
     
         17 . The device of  claim 1 , wherein microfluidic device is configured to support a cell-lysis rate of 85-100%. 
     
     
         18 . The device of  claim 1 , wherein the microfluidic device is made of a thermoplastic polymer. 
     
     
         19 . The device of  claim 18 , wherein the microfluidic device is configured to withstand high fluid pressure without deformation of the constricted regions. 
     
     
         20 . The method of  claim 19 , wherein the thermoplastic polymer comprises an off-stoichiometry thiol-ene (OSTE) polymer. 
     
     
         21 . The method of  claim 20 , wherein the OSTE polymer is made from an OSTE prepolymer having an excess of allyl groups. 
     
     
         22 . A method for continuous flow cell lysis in a microfluidic device, the method comprising:
 flowing fluid through the microfluidic device, whereby cells in the fluid are lysed,   wherein the microfluidic device comprises one or more microfluidic channels, each channel comprising constricted regions and non-constricted regions separating the constricted regions, wherein the constricted regions are configured to disrupt the cellular membranes of cells in fluid flowing through the one or more microfluidic channels.   
     
     
         23 . The method of  claim 22 , wherein the one or more microfluidic channels are parallel to each other. 
     
     
         24 . The method of  claim 22 , wherein the non-constricted regions are arranged in a honeycomb-like pattern. 
     
     
         25 . The method of  claim 22 , wherein each of the non-constricted regions has a width of 40-100 μm, and each of the constricted regions has a width of 3-10 μm. 
     
     
         26 . The method of  claim 22 , wherein each of the non-constricted regions has a length of 60-120 μm, and each of the constricted regions has a length of 10-20 μm. 
     
     
         27 . The method of  claim 22 , wherein there are 1-40 microfluidic channels. 
     
     
         28 . The method of  claim 22 , wherein each microfluidic channel comprises 3-15 constricted regions. 
     
     
         29 . The method of  claim 22 , wherein each microfluidic channel comprises a first segment of constricted regions along a fluid flow path having a width of 6-8 μm and a second segment of constricted regions along a fluid flow path having a width of 4-6 μm. 
     
     
         30 . The method of  claim 22 , wherein each of the constricted regions has a width of 2.5-4.5 μm. 
     
     
         31 . The method of  claim 22 , wherein each of the microfluidic channels has 4 constricted regions and each constricted region has a width of 3 μm. 
     
     
         32 . The method of  claim 22 , wherein there are 20 microfluidic channels. 
     
     
         33 . The method of  claim 22  wherein there are 40 microfluidic channels. 
     
     
         34 . The method of  claim 22 , wherein the microfluidic channels are configured to support a flow rate from about 20 μL/min to about 2000 μL/min. 
     
     
         35 . The method of  claim 22 , wherein the microfluidic device is configured to support a cell-lysis rate of 85-100%. 
     
     
         36 . The method of  claim 22 , wherein the microfluidic device is made of a thermoplastic polymer. 
     
     
         37 . The method of  claim 36 , wherein the microfluidic device is configured to withstand high fluid pressure without deformation of the constricted regions. 
     
     
         38 . The method of  claim 37 , wherein the thermoplastic polymer comprises an off-stoichiometry thiol-ene (OSTE) polymer. 
     
     
         39 . The method of  claim 38 , wherein the OSTE polymer is made from an OSTE prepolymer having an excess of allyl groups.