Devices and methods for determining particle concentration in a sample
A cartridge for determining a concentration of target cells within a sample includes a separation portion and a detection portion. The separation portion includes a first and second surface defining a separation chamber. The separation portion can contain a density medium having a density greater than a density of a first portion of the sample and less than a density of a second portion of the sample (that includes the target cells). The separation chamber can be fluidically coupled to an inlet reservoir such that the sample can pass from the inlet reservoir to the separation chamber during rotation. The detection portion includes a detection surface that forms a boundary of a detection chamber. The detection surface is nonparallel to the first surface such that the target cells impinge on the detection surface when passing into the detection chamber. The detection surface is configured to capture the target cells.
1 . An apparatus, comprising:
a cartridge configured to be removably coupled to a rotation element configured to rotate the cartridge about a rotation axis to capture a plurality of target cells within a sample, the cartridge comprising:
an inlet portion defining an inlet reservoir configured to contain the sample;
a separation portion including a first surface and a second surface defining a separation chamber, the separation portion configured to contain a density medium having a density greater than a density of a first portion of the sample and less than a density of a second portion of the sample, the second portion including the plurality of target cells, the separation chamber configured to be fluidically coupled to the inlet reservoir such that at least the second portion of the sample can pass from the inlet reservoir to the separation chamber when the cartridge is rotated, wherein the first surface and the second surface are parallel to each other, the first surface and the second surface each being perpendicular to a distance between the first surface and the second surface; and
a detection portion including a detection surface that forms a boundary of a detection chamber, the detection chamber fluidically coupled to the separation chamber such that at least the plurality of target cells can pass through the density medium and into the detection chamber, the detection surface being nonparallel to at least one of the first surface or the second surface such that the plurality of target cells impinge on the detection surface when passing into the detection chamber, the first surface and the detection surface forming a single bend between the first surface and the detection surface, the single bend producing a non-smooth transition from the first surface to the detection surface and defining a detection angle of between about 1 degree and about 8 degrees, the detection surface configured to capture the plurality of target cells; and
a dilution reagent, wherein at least one of the dilution reagent or the density medium comprise any of an antifoaming agent, a wetting agent, a dispersant, or an emulsifier.
2 . The apparatus of claim 1 , wherein:
the distance between the first surface and the second surface is perpendicular to an axis that intersects a central location of the cartridge in a radial direction;
the inlet portion defines an opening through which the sample can be conveyed into the inlet reservoir; and
the detection surface is angled in a downward direction with respect to the opening and bends in a downward direction with respect to the axis that intersects the central location of the cartridge in the radial direction.
3 . The apparatus of claim 1 , wherein a ratio of a volume of the separation chamber and a volume of the detection chamber is at least about 2.0.
4 . The apparatus of claim 1 , wherein the distance between the first surface and the second surface defines a thickness of the separation chamber, the thickness of the separation chamber being less than about 0.6 mm.
5 . The apparatus of claim 1 , further comprising the density medium, the density medium having a density of between 1.01 g/cm3 and 1.13 g/cm3, and wherein the cartridge includes a density medium reservoir fluidically coupled to the separation chamber.
6 . The apparatus of claim 1 , wherein at least one of the dilution reagent or the density medium comprises a poloxamer, the poloxamer containing poly(ethylene oxide) (PEO) or poly(propylene oxide) (PPO).
7 . The apparatus of claim 1 , further comprising a staining reagent, the staining reagent formulated to bind to and enhance detection of the plurality of target cells.
8 . The apparatus of claim 1 , wherein:
the cartridge further comprises a collection portion including a third surface;
the detection surface is an upper surface of the detection portion, the detection surface transitioning into the third surface where the detection portion terminates in the collection portion;
the second surface is a bottom surface of the separation portion; and
the third surface is coplanar with or lower than the second surface.
9 . An apparatus, comprising:
a cartridge configured to be removably coupled to a rotation element configured to rotate the cartridge about a rotation axis to capture a plurality of target cells within a sample, the cartridge comprising:
an inlet portion defining an inlet reservoir configured to contain the sample;
a separation portion including a first surface and a second surface defining a separation chamber, the separation portion configured to contain a density medium having a density greater than a density of a first portion of the sample and less than a density of a second portion of the sample, the second portion including the plurality of target cells, the separation chamber configured to be fluidically coupled to the inlet reservoir such that at least the second portion of the sample can pass from the inlet reservoir to the separation chamber when the cartridge is rotated; and
a detection portion including a detection surface that forms a boundary of a detection chamber, the detection chamber fluidically coupled to the separation chamber such that at least the plurality of target cells can pass through the density medium and into the detection chamber, the detection chamber being characterized by a substantially constant or diverging cross-sectional area, the detection surface being nonparallel to at least one of the first surface or the second surface such that the plurality of target cells impinge on the detection surface when passing into the detection chamber, the detection surface being angled at a non-zero angle with respect to a radial axis defined by the cartridge that is normal to the rotation axis, the detection surface configured to capture the plurality of target cells; and
a dilution reagent, wherein at least one of the dilution reagent or the density medium comprise any of an antifoaming agent, a wetting agent, a dispersant, or an emulsifier.
10 . The apparatus of claim 9 , wherein:
the first surface and the detection surface form a single bend between the first surface and the detection surface, the single bend defining a detection angle and producing a non-smooth transition from the first surface to the detection surface; and
the detection angle is between about 1 degree and about 8 degrees.
11 . The apparatus of claim 10 , wherein:
the detection angle is along the radial axis and within a cross-sectional plane defined by the radial axis and the rotation axis; and
the first surface of the separation portion and the detection surface are monolithically constructed.
12 . The apparatus of claim 10 , wherein the detection angle is about 2 degrees.
13 . The apparatus of claim 9 , wherein the detection surface includes a surface modification including a coating comprising a charged polymer to enhance adhesion of the plurality of target cells.
14 . The apparatus of claim 13 , wherein the coating is poly-L-lysine that is charged with (—NH 3 + ).
15 . The apparatus of claim 9 , wherein:
the second surface is a bottom surface of the separation portion; and
the detection surface transitions into a third surface coplanar with or lower than the second surface.
16 . The apparatus of claim 9 , wherein:
the inlet portion defines an opening through which the sample can be conveyed into the inlet reservoir; and
the detection surface is angled in an upward direction with respect to the opening.
17 . The apparatus of claim 9 , wherein:
the cartridge further comprises a hub located on the rotation axis;
the cartridge is configured to be removably coupled to the rotation element via the hub; and
a center of mass of the cartridge is located within the hub.
18 . The apparatus of claim 9 , wherein a ratio of a volume of the separation chamber and a volume of the detection chamber is at least about 2.0.
19 . An apparatus, comprising:
a cartridge configured to be removably coupled to a rotation element configured to rotate the cartridge about a rotation axis to capture a plurality of target cells within a sample, the cartridge comprising:
an inlet portion defining an inlet reservoir configured to contain the sample;
a separation portion including a first surface and a second surface defining a separation chamber, the separation portion configured to contain a density medium having a density greater than a density of a first portion of the sample and less than a density of a second portion of the sample, the second portion including the plurality of target cells, the separation chamber configured to be fluidically coupled to the inlet reservoir such that at least the second portion of the sample can pass from the inlet reservoir to the separation chamber when the cartridge is rotated, wherein the first surface and the second surface are parallel to each other, the first surface and the second surface each being perpendicular to a distance between the first surface and the second surface; and
a detection portion including a first detection surface and a second detection surface, the first detection surface and the second detection surface each being perpendicular to a distance between the first detection surface and the second detection surface, the distance between the first detection surface and the second detection surface being substantially equal to or greater than the distance between the first surface and the second surface, the first detection surface forming a first boundary of a detection chamber, the second detection surface forming a second boundary of the detection chamber, the detection chamber fluidically coupled to the separation chamber such that at least the plurality of target cells can pass through the density medium and into the detection chamber, the first detection surface and the second detection surface each being nonparallel to at least one of the first surface or the second surface such that the plurality of target cells impinge on at least one of the first detection surface or the second detection surface when passing into the detection chamber, at least one of the first detection surface or the second detection surface configured to capture the plurality of target cells; and
a dilution reagent, wherein at least one of the dilution reagent or the density medium comprise any of an antifoaming agent, a wetting agent, a dispersant, or an emulsifier.
20 . The apparatus of claim 19 , wherein:
the distance between the first surface and the second surface is perpendicular to an axis that intersects a central location of the cartridge in a radial direction;
the inlet portion defines an opening through which the sample can be conveyed into the inlet reservoir; and
at least one of the first detection surface or the second detection surface is angled in a downward direction with respect to the opening and bends in a downward direction with respect to the axis that intersects the central location of the cartridge in the radial direction.
21 . The apparatus of claim 19 , further comprising the density medium, the density medium having a density of between 1.01 g/cm3 and 1.13 g/cm3, and wherein the cartridge includes a density medium reservoir fluidically coupled to the separation chamber.
22 . An apparatus, comprising:
a cartridge configured to be removably coupled to a rotation element configured to rotate the cartridge about a rotation axis to capture a plurality of target cells within a sample, the cartridge comprising:
an inlet portion defining an inlet reservoir configured to contain the sample;
a separation portion including a first surface and a second surface defining a separation chamber, the second surface being a bottom surface of the separation portion, the separation portion configured to contain a density medium having a density greater than a density of a first portion of the sample and less than a density of a second portion of the sample, the second portion including the plurality of target cells, the separation chamber configured to be fluidically coupled to the inlet reservoir such that at least the second portion of the sample can pass from the inlet reservoir to the separation chamber when the cartridge is rotated; and
a detection portion including a detection surface that forms a boundary of a detection chamber, the detection chamber fluidically coupled to the separation chamber such that at least the plurality of target cells can pass through the density medium and into the detection chamber, the detection surface being nonparallel to at least one of the first surface or the second surface such that the plurality of target cells impinge on the detection surface when passing into the detection chamber, the detection surface being angled at a non-zero angle with respect to a radial axis defined by the cartridge that is normal to the rotation axis, the detection surface configured to capture the plurality of target cells, the detection surface transitioning into a third surface coplanar with or lower than the second surface; and
a dilution reagent, wherein at least one of the dilution reagent or the density medium comprise any of an antifoaming agent, a wetting agent, a dispersant, or an emulsifier.
23 . The apparatus of claim 22 , further comprising the density medium, the density medium having a density of between 1.01 g/cm3 and 1.13 g/cm3, and wherein the cartridge includes a density medium reservoir fluidically coupled to the separation chamber.