High-density cell microarrays for parallel functional determinations

• US 20050014155A1
• Filed: 11/15/2002
• Published: 01/20/2005
• Est. Priority Date: 11/16/2001
• Status: Abandoned Application

• Current Assignee: Memorial Sloan-Kettering Cancer Center
• Original Assignee: Memorial Sloan-Kettering Cancer Center
• Inventors: Xu, Wilson C
• Application Number:
  US10/495,521
• Publication Number:
  US 20050014155A1
• Time in Patent Office: Days
• US Class Current: 506/10
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00382 Stamping
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00603 Making arrays on substantially continuous surfaces
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 35/00029 provided with flat sample substrates, e.g. slides G01N35/028 takes precedence

Abstract

Disclosed are methods for generating high-density cell microarrays. The methods generally involve forming nanocraters on a permeable membrane surface and inoculating the nanocraters with cells, proteins, or other molecules. The high-density microarrazs of the invention are useful for large-scale, high throughput phenotypic determinations of gene activities.

Claims

1-66. -66. (Canceled)

67. A cell microarray comprising a plurality of cell colonies on a membrane surface, wherein said microarray has a density of at least 5 colony spots/mm².

68. The cell microarray of claim 67, wherein said microarray has a density of at least 100 colony spots/mm².

69. The cell microarray of claim 68, wherein said microarray has a density of about 1,000 colony spots/mm².

70. The cell microarray of claim 67, wherein said membrane is a permeable, flexible membrane having a plurality of nanocraters on its surface and at least one cell within said nanocraters.

71. The microarray of claim 70, wherein said membrane is a cellulose ester membrane.

72. The microarray of claim 70, wherein the distance between the centers of adjacent nanocraters is between 400 μ

m and 200 μ

m.

73. The microarray of claim 72, wherein the distance between the centers of adjacent nanocraters is about 375 μ

m.

74. The microarray of claim 70, wherein said nanocraters have a volume between about 1.5 nano-liters and about 100 pico-liters.

75. The microarray of claim 70, wherein said nanocraters are less than 100 pico-liters in size.

76. The microarray of claim 70, wherein said nanocraters have a diameter of 125 μ

m or less.

77. The microarray of claim 70, wherein said nanocraters have a diameter of about 125 μ

m.

78. The microarray of claim 70, wherein said nanocraters have a depth of between about 10 μ

m and about 125 μ

m.

79. The microarray of claim 67, wherein said cell is from an organism selected from the group consisting of bacteria, fungi, protozoa, and algae.

80. The microarray of claim 67, wherein said cell is mammalian.

81. The microarray of claim 67, wherein said microarray comprises at least two different cell types.

82. The microarray of claim 81, wherein said cell types comprise cells of the same genus and species.

83. The method of claim 81, wherein said cell types comprise cells that differ in one or more genes.

84. A method of identifying a drug target, said method comprising the steps of:

(a) providing a membrane surface comprising a plurality of nanocraters, wherein at least one nanocrater contains a first cell type and at least one nanocrater contains a second cell type;

(b) exposing said membrane to a test substrate;

(c) detecting the response of said first cell type and said second cell type to said test substrate; and

(d) comparing the response of said first cell type and said second cell type.

85. A method of determining the function of a gene, said method comprising the steps of:

(a) providing a membrane surface comprising a plurality of nanocraters, wherein at least one nanocrater contains a first cell type and at least one nanocrater contains a second cell type, wherein said first and second cell types differ in at least one gene;

(b) exposing said membrane to a test substrate;

(c) detecting the response of said first cell type and said second cell type to said test substrate; and

(d) comparing the response of said first cell type and said second cell type.