Laminar fluidic separation in flowcells for minimal reagent usage

Patent metadata

Field	Value
Publication number	US-2017259262-A1
Application number	US-201715447664-A
Country	US
Kind code	A1
Filing date	Mar 2, 2017
Priority date	Mar 9, 2016
Publication date	Sep 14, 2017
Grant date	—

Abstract

A processing instrument flowcell having a flowcell channel with an upstream channel end, a downstream channel end, a longitudinal axis extending from the upstream channel end to the downstream channel end, and a first operative surface extending between the upstream channel end and the downstream channel end and configured to receive a first number of DNA templates. A first reagent inlet is fluidically connected to the upstream channel end at a location adjacent the first operative surface. A buffer inlet is fluidically connected to the upstream channel end at a location spaced from the first operative surface. An outlet fluidically connected to the downstream channel end. Also provided is a method for operating a flowcell channel under laminar flow conditions to maintain a first reagent adjacent a first operative surface and a buffer fluid spaced from the first operative surface. The flowcell channel may have multiple separate operative surfaces.

First claim

We claim:

1 . A flowcell for a processing instrument, the flowcell comprising:
a flowcell channel extending along a longitudinal axis from an upstream channel end to a downstream channel end, and having a first operative surface extending between the upstream channel end and the downstream channel end, the first operative surface being configured to receive a first plurality of DNA templates; a first reagent inlet fluidically connected to the upstream channel end at a first location adjacent the first operative surface; a buffer inlet fluidically connected to the upstream channel end at a second location spaced from the first operative surface; an outlet fluidically connected to the downstream channel end.

2 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height in a direction perpendicular to the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced a predetermined distance from the first operative surface in the direction perpendicular the longitudinal axis; and the predetermined distance is equal to 0.4% to 50% of the predetermined height.

3 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height in a direction perpendicular to the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced a predetermined distance from the first operative surface in the direction perpendicular the longitudinal axis; and the predetermined distance is equal to 1.4% to 18% of the predetermined height.

4 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height in a direction perpendicular to the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced a predetermined distance from the first operative surface in the direction perpendicular the longitudinal axis; and the predetermined distance is equal to 3.6% to 10.7% of the predetermined height.

5 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height of about 140 micrometers in a direction perpendicular the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced about 0.5 micrometers to about 50 micrometers from the first operative surface in the direction perpendicular the longitudinal axis.

6 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height of about 140 micrometers in a direction perpendicular the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced about 2 micrometers to about 25 micrometers from the first operative surface in the direction perpendicular the longitudinal axis.

7 . The flowcell of claim 1 , wherein:
the upstream channel end has a predetermined height of about 140 micrometers in a direction perpendicular the longitudinal axis; the first reagent inlet comprises an exit portion adjacent to the upstream channel end, and the exit portion extends from a first point adjacent to the first operative surface to a second point that is spaced about 5 micrometers to about 15 micrometers from the first operative surface in the direction perpendicular the longitudinal axis.

8 . The flowcell of claim 1 , wherein:
the first reagent inlet comprises a first reagent inlet exit portion located immediately upstream of the upstream channel end, and the exit portion is parallel to and coincident with the first operative surface; and the buffer inlet comprises a buffer inlet exit portion located immediately upstream of the upstream channel end, and the buffer inlet exit portion is parallel to and spaced from the first operative surface.

9 . The flowcell of claim 1 , wherein the first operative surface comprises a transparent material.

10 . The flowcell of claim 1 , wherein:
the flowcell channel further comprises a second operative surface extending between the upstream channel end and the downstream channel end and configured to receive a second plurality of DNA templates; the flowcell further comprises a second reagent inlet fluidically connected to the upstream channel end at a third location adjacent the second operative surface; and wherein the second location is spaced from the second operative surface.

11 . The flowcell of claim 10 , wherein the first operative surface is parallel to and facing the second operative surface.

12 . The flowcell of claim 10 , wherein the first reagent inlet and the second reagent inlet are fluidically connected at a location upstream of the flowcell channel.

13 . A method of operating a processing instrument, the method comprising:
providing a flowcell channel extending along a longitudinal axis from an upstream channel end to a downstream channel end, and having a first operative surface extending between the upstream channel end and the downstream channel end, the first operative surface comprising a first plurality of DNA templates; providing a first reagent fluid to the upstream channel end channel at a first location adjacent the first operative surface; providing a barrier fluid that is different from the first reagent fluid to the upstream channel end at a second location spaced from the first operative surface; and passing the first reagent fluid and the barrier fluid through the flowcell channel under laminar flow conditions such that the first reagent fluid remains adjacent the first operative surface and the barrier fluid remains spaced from the first operative surface from the upstream channel end to the downstream channel end.

14 . The method of claim 13 , wherein the barrier fluid remains spaced from the first operative surface from the upstream channel end to the downstream channel end by a distance equal to 0.4% to 50% of a total height of the flowcell channel at the upstream channel end as measured between the first operative surface and an opposite interior wall of the flowcell channel.

15 . The method of claim 13 , wherein the barrier fluid remains spaced from the first operative surface from the upstream channel end to the downstream channel end by a distance equal to 1.4% to 18% of a total height of the flowcell channel at the upstream channel end as measured between the first operative surface and an opposite interior wall of the flowcell channel.

16 . The method of claim 13 , wherein the barrier fluid remains spaced from the first operative surface from the upstream channel end to the downstream channel end by a distance equal to 3.6% to 10.7% of a total height of the flowcell channel at the upstream channel end as measured between the first operative surface and an opposite interior wall of the flowcell channel.

17 . The method of claim 13 , wherein a total height of the flowcell channel at the upstream channel end as measured between the first operative surface and an opposite interior wall of the flowcell

Assignees

• Intelligent Bio-Systems Inc
• Qiagen Gmbh

Inventors

• Perroud Thomas Daniel
• Perbost Michel Georges
• Focke Maximilian

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• B01L2200/16

  Reagents, handling or storing thereof · CPC title

• B01L2300/168

  Specific optical properties, e.g. reflective coatings · CPC title

• B01L2200/0673

  Handling of plugs of fluid surrounded by immiscible fluid · CPC title