Device for analysis of cellular motility

Abstract

A mesoscale fluidic system comprises a substrate having a sample chamber and an analysis chamber. The sample chamber comprises a cell permeable filter defining a sample application compartment and a conditioning medium compartment. The sample chamber has a sample inlet port in the sample application compartment. The analysis chamber has an entry port and an exit port. The conditioning medium compartment is in fluid communication with the entry port of the analysis chamber via a channel. The sample application compartment is below the cell permeable filter and the conditioning medium compartment is above the cell permeable filter. The mesoscale fluidic system is suited for analysing cellular motility in a sample. Also disclosed is a method of estimating the quantity of motile cells in a sample and a method of extracting motile cells from non-motile cells.

1 - 34 . (canceled) 
     
     
         35 . A mesoscale fluidic system for separating motile cells from non-motile cells, the system comprising:
 a substrate having a sample chamber and an analysis chamber,   the sample chamber comprising a cell permeable filter defining a sample application compartment and a conditioning medium compartment, which cell permeable filter has a pore size in the range of from 1 μm to 20 μm,   the sample chamber having a sample inlet port in the sample application compartment,   the analysis chamber having an entry port and an exit port,   the conditioning medium compartment being in fluid communication with the entry port of the analysis chamber via a channel and the conditioning medium compartment further comprising a medium inlet port in fluid communication with the ambient;   wherein with reference to the gravitational field the sample application compartment is below the cell permeable filter and the conditioning medium compartment is above the cell permeable filter.   
     
     
         36 . The mesoscale fluidic system according to  claim 35 , wherein the analysis chamber comprises a cell retaining filter having an upstream surface facing the entry port and a downstream surface facing the exit port, which cell retaining filter has a pore size that retains the motile cells, which pore size is lower than the pore size of the cell permeable filter and is in the range of from 0.1 μm to 12 μm. 
     
     
         37 . The mesoscale fluidic system according to  claim 36 , wherein at least a portion of the substrate is transparent allowing that at least one of the upstream surface and the downstream surface of the cell retaining filter is observed visually. 
     
     
         38 . The mesoscale fluidic system according to  claim 35 , wherein the analysis chamber comprises a closable member allowing physical access to the analysis chamber. 
     
     
         39 . The mesoscale fluidic system according to  claim 35 , further comprising a device to provide a liquid driving force to move a liquid from the sample chamber to the analysis chamber or from the analysis chamber to the sample chamber. 
     
     
         40 . The mesoscale fluidic system according to  claim 39 , wherein the device to provide a liquid driving force comprise a syringe. 
     
     
         41 . The mesoscale fluidic system according to  claim 35 , further comprising a reservoir in fluid communication with the sample application compartment, the conditioning medium compartment or the analysis chamber. 
     
     
         42 . The mesoscale fluidic system according to  claim 35 , further comprising a receiving well in fluid communication with the sample inlet port. 
     
     
         43 . The mesoscale fluidic system according to  claim 35 , wherein the channel has a smallest dimension in the range of from about 10 μm to about 4 mm. 
     
     
         44 . The mesoscale fluidic system according to  claim 35 , wherein at least one of the sample chamber and the analysis chamber have a depth in the range of from about 100 μm to about 20 mm. 
     
     
         45 . The mesoscale fluidic system according to  claim 36 , wherein cell retaining filter has a pore size in the range of from 0.1 μm to 1.0 μm. 
     
     
         46 . The mesoscale fluidic system according to  claim 35 , further comprising a sample medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents, a conditioning medium comprising at least one of nutrients, salts, buffers, and at least one of viscosity modifying agents and a detection agent. 
     
     
         47 . A method of separating motile cells from non-motile cells in a sample comprising the steps of:
 providing a cell permeable filter having a pore size allowing the motile cells to move through the cell permeable filter;   applying a sample containing motile cells below the cell permeable filter;   allowing motile cells in the sample to move through the cell permeable filter, and   detecting motile cells having moved through the cell permeable filter, or   extracting motile cells having moved through the cell permeable filter.   
     
     
         48 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , wherein in the step of detecting motile cells having moved through the cell permeable filter the motile cells are detected using a detection agent. 
     
     
         49 . The method of separating motile cells from non-motile cells in a sample according to  claim 48 , wherein the detection agent is a tetrazolium dye, such as MTT. 
     
     
         50 . The method of separating motile cells from non-motile cells in a sample according to  claim 48 , wherein in the step of detecting motile cells having moved through the cell permeable filter two or more detection agents are used. 
     
     
         51 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , further comprising the step of applying a sample medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents below the cell permeable filter. 
     
     
         52 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , further comprising the step of applying a cell conditioning medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents above the cell permeable filter. 
     
     
         53 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , wherein the cell permeable filter has a pore size in the range of from 1 μm to 20 μm. 
     
     
         54 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , wherein the motile cells are eukaryotic cells, such as mammalian sperm cells. 
     
     
         55 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , wherein the motile cells are prokaryotic cells. 
     
     
         56 . The method of separating motile cells from non-motile cells in a sample according to  claim 47 , wherein the cell permeable filter is contained in a mesoscale fluidic system according to  claim 35 . 
     
     
         57 . The method of separating motile cells from non-motile cells in a sample according to  claim 56  further comprising the steps of:
 adding a conditioning medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents to the sample chamber and the analysis chamber and the channel between them; 
 optionally adding a detection agent to the sample chamber; 
 adding the sample to the sample application compartment; 
 allowing the motile cells of the sample to move to the conditioning medium compartment; 
 providing a liquid driving force from the sample chamber to the analysis chamber; 
 quantifying the detection agent in the analysis chambers or extracting the motile cells from the analysis chamber. 
 
     
     
         58 . A kit of parts comprising a mesoscale fluidic system according to  claim 35 , a detection agent, a sample medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents and a cell conditioning medium comprising at least one of nutrients, salts, buffers, and viscosity modifying agents. 
     
     
         59 . A kit of parts according to  claim 58 , wherein the detection agent is a tetrazolium dye. 
     
     
         60 . A computer implemented method for determining a value of a quantifiable measure of cell motility for a cell sample comprising the steps of:
 receiving image data comprising an image of at least a part of at least one of an analysis chamber and a cell retaining filter of a mesoscale fluidic system;   processing the image data to determine the value of the quantifiable measure;   wherein the step of processing the image data comprises the step of mapping the image data into the value of the quantifiable measure using a mapping function, the mapping function being calibrated to map image data of at least one of an analysis chamber and a cell retaining filter of a mesoscale fluidic system according to  claim 35  into a value of the quantifiable measure.   
     
     
         61 . A computer implemented method according to  claim 60 , further comprising the step of pre-processing the image data to determine whether the image data is usable as input to the mapping function, wherein an error message is generated if it is determined that the image data is un-usable as input. 
     
     
         62 . A computer implemented method according to  claim 60 , further comprising the step of:
 displaying the determined value of the quantifiable measure on a display.   
     
     
         63 . A computer program product comprising program code adapted to cause a data processing system to perform the steps of the computer implemented method according to  claim 60 , when the program code is executed on the data processing system, wherein the computer program product comprises a computer-readable medium having stored thereon the program code. 
     
     
         64 . A device for determining a value of a quantifiable measure of cell motility for a cell sample, the device comprising:
 a camera unit for capturing an image of at least one of a part of an analysis chamber and a cell retaining filter of a mesoscale fluidic system, a processing unit for processing the captured image, a display for displaying the value of the a quantifiable measure of cell motility, and a user input unit, wherein, in response to activation of the user input unit;   the camera unit is configured to capture an image of at least one of a part of an analysis chamber and a cell retaining filter of a mesoscale fluidic system;   the processing unit is configured to process the captured image to determine the value of the quantifiable measure of cell motility using a mapping function configured to map the captured image into the value of the quantifiable measure, wherein the mapping function is calibrated to map image data of at least one of an analysis chamber and a cell retaining filter of a mesoscale fluidic system according to  claim 35  into a value of the quantifiable measure.   
     
     
         65 . A data storage medium comprising a computer readable code, wherein said computer readable code is configured to program a user terminal comprising a processing unit, a display, and a user input unit to become a device according to  claim 64 .