Digital fluid sample separation apparatus and methods for one-step quantitative sample analysis

A gas permeable apparatus for automatically separating, digitizing, compartmentalizing and analyzing a fluid sample, the apparatus comprising:

• (a) a fluidic layer configured to separate a fluid sample into wells for fluid sample analysis, said fluidic layer comprising:
• (i) a plurality of wells;
• (ii) a sample inlet that receives the fluid sample;
• (iii) a main channel that transports the fluid sample from the sample inlet to one or more wells, wherein flow of the fluid sample occurs automatically by vacuum diffusion;
• (iv) a plurality of cross channels each individually coupling the main channel with a dedicated well from among the plurality of wells, wherein the cross channels have a cross-sectional area smaller than a cross-sectional area of the wells and the cross-sectional area of the main channel so as to efficiently compartmentalize the fluid sample into the plurality of wells and prevent backflow of the fluid sample from re-entering the main channel from the wells, wherein the wells hold the compartmentalized fluid sample for multiplexed analysis via distinct and independent reactions within each well or distinct and independent targets within each well, wherein the multiplexed fluid sample analysis comprises detection of specific molecules within the fluid sample, wherein molecules to be amplified before detection are automatically amplified in the wells before detection;
• and
• (b) a cover layer configured to seal the fluidic layer;
• (c) wherein at least one of the fluidic layer and cover layer comprise a gas permeable material for allowing the fluid sample to flow automatically into the wells by vacuum diffusion;
• and
• (d) wherein the main channel, plurality of cross channels and plurality of wells have a shape and size to allow for loading, separation and compartmentalization of the fluid sample into the wells in less than 10 minutes.

An apparatus for separating, digitizing and analyzing a fluid sample, the apparatus comprising:

• (a) a fluidic layer configured to separate a fluid sample into wells for fluid sample analysis, said fluidic layer comprising:
• (i) a plurality of wells;
• (ii) a sample inlet that receives the fluid sample;
• (iii) a main channel that transports the fluid sample from the sample inlet to one or more wells;
• (iv) a plurality of cross channels each individually coupling the main channel with a dedicated well from among the plurality of wells, wherein the cross channels have a cross-sectional area smaller than a cross-sectional area of the main channel and a cross-sectional area of the wells so as to efficiently compartmentalize the fluid sample into the plurality of wells and prevent backflow of the fluid sample from re-entering the main channel from the wells;
• (v) wherein the wells hold the compartmentalized fluid sample for multiplexed analysis via distinct and independent reactions within each well or distinct and independent targets within each well;
• and
• (b) a cover layer configured to seal the fluidic layer;
• (c) wherein at least one of the fluidic layer and cover layer comprise a gas permeable material, allowing the fluid sample to flow automatically into the wells by vacuum diffusion.