Integrated device for comprehensive tissue to single-cell sample preparation

A method for preparing a complex tissue sample into single cells for direct single-cell analysis (SCA), the method comprising the steps of:

• (1) obtaining a device comprising:
• a pump capable of providing a back and forth liquid flow including a forward flow and a reverse flow through the device;
• a tissue input well for holding the tissue sample, said well in fluid communication with an inlet comprising a first mesh and an outlet comprising a second mesh;
• wherein the first mesh is operative to retain the tissue sample in the well when a reverse flow is established from the outlet to the inlet, and the second mesh is operative to retain the tissue sample in the input well when a forward flow is established from the inlet to the outlet yet allow clumps of cells, cell aggregates, or single cells through the input well's outlet;
• at least one microfluidic channel in fluid communication with the outlet with an optional third mesh disposed therein the microfluidic channel;
• and
• an acoustic inlet in fluid communication with the at least one microfluidic channel, connected to a first acoustic region comprising a high frequency acoustic transducer operative to direct acoustic (sound) waves to clumps of cells in the acoustic region, which comprises a fourth mesh at a first acoustic outlet in fluid communication with a microchannel;
• wherein the device is capable of receiving flow from a pump from the inlet to the outlet, then the pump repeatedly reverses, and flow is directed from the outlet to the inlet and vice versa, causing a liquid flow with the repeated back and forth flow through the device, thereby eluting purified single cells, suitable for SCA, from the microchannel of the acoustic outlet;
• (2) disposing a tissue sample into the tissue input well;
• (3) establishing an acoustic (sound) energy in the device and at least one of a liquid flow through the device and an electrical field in the device;
• (4) providing a back and forth liquid flow including a forward flow and a reverse flow through the device;
• whereby the device receives flow from the pump from the inlet to the outlet, then the pump repeatedly reverses, and flow is directed from the outlet to the inlet and vice versa, causing a liquid flow with the repeated back and forth flow through the device, thereby eluting purified single cells, suitable for SCA, from the microchannel of the acoustic outlet.

A method for diagnosing a condition in a subject in need thereof, the method comprising the steps of:

• (1) obtaining a device comprising:
• a pump capable of providing a back and forth liquid flow including a forward flow and a reverse flow through the device;
• a tissue input well for holding the tissue sample, said well in fluid communication with an inlet comprising a first mesh and an outlet comprising a second mesh;
• wherein the first mesh is operative to retain the tissue sample in the well when a reverse flow is established from the outlet to the inlet, and the second mesh is operative to retain the tissue sample in the input well when a forward flow is established from the inlet to the outlet yet allow clumps of cells, cell aggregates, or single cells through the input well's outlet;
• at least one microfluidic channel in fluid communication with the outlet with an optional third mesh disposed therein the microfluidic channel;
• and
• an acoustic inlet in fluid communication with the at least one microfluidic channel, connected to a first acoustic region comprising a high frequency acoustic transducer operative to direct acoustic (sound) waves to clumps of cells in the acoustic region, which comprises a fourth mesh at a first acoustic outlet in fluid communication with a microchannel;
• wherein the device is capable of receiving flow from a pump from the inlet to the outlet, then the pump repeatedly reverses, and flow is directed from the outlet to the inlet and vice versa, causing a liquid flow with the repeated back and forth flow through the device, thereby eluting purified single cells, suitable for SCA, from the microchannel of the acoustic outlet;
• (2) disposing a tissue sample into the tissue input well;
• (3) establishing an acoustic (sound) energy in the device and at least one of a liquid flow through the device and an electrical field in the device;
• (4) providing a back and forth liquid flow including a forward flow and a reverse flow through the device;
• whereby the device receives flow from the pump from the inlet to the outlet, then the pump repeatedly reverses, and flow is directed from the outlet to the inlet and vice versa, causing a liquid flow with the repeated back and forth flow through the device, thereby eluting purified single cells, suitable for SCA, from the microchannel of the acoustic outlet;
• and comprising the steps of:
• (A) a candidate tissue sample is provided from the subject;
• (B) cells suitable for SCS are provided by the method of steps (1)-(4) above;
• (C) analyzing/detecting at least a single cell from the subject, said single cell indicative of the condition;
• and
• (D) reporting a cell type of the single cell;
• whereby the cell type reported is indicative of or diagnostic of a condition of the subject.