# US 2014/0193900 A1 · Device for washing suspended cells or particles

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- Number: US 2014/0193900 A1
- Document ID: US20140193900A1
- Status: Published application
- Kind: A1
- Application number: 14/124,900
- Filed: 2012-05-29
- Published: 2014-07-10
- Earliest claimed: 2011-06-09
- Inventors: Ivar Meyvantsson; Steve Hayes; Victoria Echeverria; Robert Lowery; Casey Lamers
- Web page: https://patentgrep.com/patent/US20140193900A1

## CPC

- C12M 47/12: {Purification (C12M47/04 takes precedence)}
- B01L 3/5085: {for multiple samples, e.g. microtitration plates}
- C12M 47/04: {Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04)}
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- B01L 2200/0668: Trapping microscopic beads
- B01L 2300/0829: Multi-well plates; Microtitration plates
- B01L 2400/086: using baffles or other fixed flow obstructions
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A device and method are disclosed to enable fluid manipulations without disturbing or removing particles or cells. By engineering assay compartment geometries to take advantage of fluid mechanical effects, the flow velocity may be diminished around particles or cells. This technology may be used to process particles or cells of various kinds that are suspended in liquids or gels, and is suitable for use in high throughput screening.

## Claims

### Claim 1

A device with patterned structures disposed to selectively diminish convective fluid flow, the device comprising:
at least one well configured to receive particles or cells, the at least one well having a bottom with at least one portion containing patterned structures disposed to allow the particles or cells to settle out of liquid suspension onto the lowest surface within the well,
wherein the at least one well is disposed to enable fluid delivery and removal in a volume surrounding the patterned structures and the patterned structures are disposed to diminish fluid flow at the lowest surface of the patterned structures thereby protecting the particles or cells residing thereon from fluid flow forces when fluid is delivered or removed from the volume surrounding the patterned structures.

### Claim 2 (depends on 1)

The device of claim 1 wherein the patterned structures are either protrusions or depressions.

### Claim 3 (depends on 1)

The device of claim 1 further comprising at least one shelf at the bottom of the well, where the at least one shelf is elevated above the bottom portion of the well, where the at least one shelf is connected to a sidewall of the well, where the at least one shelf divides the well into two or more segments, or where the at least one shelf is not connected to the sidewall of the well.

### Claim 4 (depends on 3)

The device of claim 3 wherein the ratio of the area divided between the at least one shelf and the patterned structures is in a range of 1:100,000 to 100,000:1.

### Claim 5 (depends on 3)

The device of claim 3 wherein the ratio of the area divided between the at least one shelf and the patterned structures is in a range of 1:100 to 100:1.

### Claim 6 (depends on 3)

The device of claim 3 wherein the ratio of the area divided between the at least one shelf and the patterned structures is in a range of 1:20 to 20:1.

### Claim 7 (depends on 1)

The device of claim 1 wherein the at least one well ranges in size from those within a standard 3456-well plate (0.5-2 mm length/width/diameter by 3-4 mm height) to those within a standard bioassay dish (200-250 mm length/width by 20-30 mm height).

### Claim 8 (depends on 1)

The device of claim 1 wherein the at least one well ranges in size from those within a standard 384-well plate (3-4 mm length/width/diameter by 11-12 mm height) to those within a standard petri dish (30-150 mm diameter by 12-20 mm height).

### Claim 9 (depends on 1)

The device of claim 1 wherein the at least one well are square or round and are sized equivalent to those within a standard 384-well plate (3-4 mm length/width/diameter by 11-12 mm height).

### Claim 10 (depends on 1)

The device of claim 1 further comprising macro and micro structures within the at least one well, wherein the macro and micro structures occur as arrays in the forms of linearly or offset arranged posts, linear baffles or walls, intersecting walls, curved baffles or walls, or with microwells or holes.

### Claim 11 (depends on 1)

The device of claim 1 wherein the macro or micro structures within the at least one well have an aspect ratio of height to width within the range of 1:1000 to 1000:1.

### Claim 12 (depends on 1)

The device of claim 1 wherein the macro or micro structures within the at least one well have an aspect ratio of height to width within the range of 1:100 to 100:1.

### Claim 13 (depends on 1)

The device of claim 1 wherein the macro or micro structures within the at least one well have an aspect ratio of height to width within the range of 1:10 to 10:1.

### Claim 14 (depends on 1)

The device of claim 1 wherein the macro or micro structures within the at least one well have an aspect ratio of height to width within the range of 1:3 to 3:1.

### Claim 15 (depends on 1)

A method for using the device of claim 1 comprising in any order, combination or number of permutations:
providing a volume of liquid containing particles or cells to the at least one well;
providing an additional volume of liquid;
allowing the particles or cells to settle to the lowest surface of the at least one well by gravity, centrifugation, affinity or magnetism; and
performing a wash cycle at least once, said wash cycle comprising removing an excess fluid volume and replacing at least a part of this removed fluid volume.

### Claim 16 (depends on 15)

The method of claim 15 wherein the rate of dispensing and removing fluid volume is in the range of 10 nl/sec to 1 ml/sec.

### Claim 17 (depends on 15)

The method of claim 15 wherein the rate of dispensing and removing fluid volume is in the range of 50 nl/sec to 100 μl/sec.

### Claim 18 (depends on 15)

The method of claim 15 wherein the rate of dispensing and removing fluid volume is in the range of 100 nl/sec to 10 μl/sec.

### Claim 19 (depends on 15)

The method of claim 15 wherein a loss of cells or particles following the wash cycle is less than 10% of the cells or particles present.

### Claim 20 (depends on 15)

The method of claim 15 wherein a loss of cells or particles following 10 wash cycles is less than 10% of the cells or particles present.

### Claim 21 (depends on 15)

The method of claim 15 wherein a loss of cells or particles following 15 wash cycles is less than 10% of the cells or particles present.

### Claim 22 (depends on 15)

The method of claim 15 wherein the concentration of a dissolved substance present after the wash cycle is less than 50% of the concentration of the dissolved substance before the wash cycle.

### Claim 23

A device for selectively diminishing convective fluid flow, the device comprising:
at least one well configured to receive particles or cells, the at least one well having a bottom with at least one portion disposed to allow the particles or cells to settle out of liquid suspension onto a lowest surface of the well,
wherein the at least one well is disposed to enable fluid delivery and removal in a volume surrounding the bottom portion of the well and the bottom portion is disposed to diminish fluid flow at the lowest surface thereof, thereby protecting the particles or cells residing thereon from fluid flow forces when fluid is delivered or removed from the volume surrounding the bottom of the well.