Affinity binding-based system for detecting particulates in a fluid
AssigneeGamera Bioscience Corporation
InventorsNorman F. Sheppard, Jr., Alec Mian, Gregory Kellogg, Stephen G. Kieffer-Higgins, Bruce L. Carvalho
Life of this patent
Estimating the term…
Abstract
This invention provides methods an apparatus for detecting and quantifying particulate matter suspended in a fluid. Specifically, the invention provides an integrated, affinity-binding based, analytical system comprising a platform for performing an affinity-binding based assay for specifically binding particulates including microbial cells, and a detection means for detecting the particulates specifically bound to a defined surface or chamber comprising the platform. Methods for using the analytical systems of the invention are also provided.
Claims40
A centripetally-motivated apparatus for detecting a cell in a fluid, the apparatus comprising
- a platform, comprising a substrate having a first flat, planar surface and a second flat, planar surface opposite thereto, wherein the first surface defines a cell accumulation chamber wherein a defined amount of a fluid comprising cells is retained in the chamber for a time sufficient to permit the cells to accumulate in the chamber, and further comprises a multiplicity of microchannels embedded within the first planar surface and a sample input means, wherein the cell accumulation chamber, the sample input means and the microchannels are connected and in fluid communication, aid wherein the platform further comprises
- a wash buffer reservoir containing a wash buffer in fluid communication with the cell accumulation chamber;
- a fluid waste receptacle in fluid communication with the cell accumulation chamber, wherein the cell accumulation chamber is positioned radially more distal from the center of the platform than the sample input means or the wash buffer reservoir and the microchannels are arrayed between the sample input means, the cell accumulation chamber, the wash buffer reservoir and the fluid waste receptacle to provide fluid communication there between;
- the platform comprising the apparatus in combination with
- a device, comprising a base, a rotating means, a power supply and user interface and operations controlling means, wherein the rotating means is operatively linked to the platform and in rotational contact therewith
- wherein a fluid sample comprising cells is moved from the sample input means to the cell accumulation chamber of the platform by centripetal force arising from rotational motion of the platform for a time and a rotational velocity sufficient to move the fluid through the microchannels, and is incubated thereon for a time sufficient to permit the cells to accumulate in the area;
- and
- wherein the fluid sample is replaced with the wash buffer and displaced into the fluid waste receptacle by centripetal force arising from rotational motion of the platform for a time and a rotational velocity sufficient to move the fluid through the microchannels;
- and
- wherein the wash buffer is further displaced into the fluid waste receptacle by centripetal force arising from rotational motion of tile platform for a time and a rotational velocity sufficient to move the fluid through the microchannels;
- and
- wherein a cell in the cell accumulation chamber is detected thereupon.
On a platform comprising an analytical apparatus for detecting a particulate in a fluid sample, a detection chamber comprising a cell adhesion agent wherein the detection chamber has a patterned reflective surface comprising an optically transparent portion and a reflective portion wherein the transparent portion of the surface is coated with the cell adhesion agent and the reflective portion of the surface is coated with a blocking agent that prevents non-specific binding thereto.
Dependencies are inferred from the claim text.