Capture and detection of microbes by membrane methods

• US 20060079000A1
• Filed: 01/24/2005
• Published: 04/13/2006
• Est. Priority Date: 07/24/2002
• Status: Abandoned Application

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: McDevitt, John T., Floriano, Pierre, Christodoulides, Nick
• Application Number:
  US10/522,499
• Publication Number:
  US 20060079000A1
• Time in Patent Office: Days
• US Class Current: 436/164
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/22 Testing for sterility conditions
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 1/4077 by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 15/0227 using imaging; using holography
• CPC Class Codes: G01N 15/0272 with screening; with classification by filtering B01D takes precedence
• CPC Class Codes: G01N 2015/019 Biological contaminants; Fouling
• CPC Class Codes: G01N 33/56911 Bacteria

Abstract

Methods and systems for detecting the presence of analytes using a membrane based detection system are described. A fluid sample is passed through a membrane based detection system (100). Particulate analytes (e.g., microbes) are captured by the membrane (110). Detection and analysis techniques may be applied to determine the identity and quantity of the captured analytes.

Claims

1-49. -49. (canceled)

50. A method of analyzing an analyte collected on a membrane comprising:

passing a fluid sample across a membrane, wherein the fluid sample comprises an analyte that is at least partially retained by the membrane;

adding a visualization agent to material collected on the membrane when the fluid sample is passed across the membrane;

collecting an image of the collected material using white light, at a first wavelength of light, a second wavelength of light, and a third wavelength of light, wherein the analyte comprises a color corresponding to the first wavelength of light;

forming a first mask corresponding to an image of the collected material at the second wavelength of light;

forming a second mask corresponding to an image of the collected material at the third wavelength of light;

subtracting the first mask and the second mask from the image of the collected material in white light.

51. The method of claim 50, wherein the wavelengths of light are selected from the group consisting of red, blue and green.

52. The method of claim 50, wherein the collecting the image data and forming the masks is performed by a computer.

53. The method of claim 50, further comprising determining the amount of analyte present on the membrane by analysis of the image resulting from subtracting the first mask and the second mask from the image of the collected material in white light.

54. The method of claim 50, wherein the images are collected using a digital detection device.

81. The method of claim 50, wherein the membrane is coupled to a body.

82. The method of claim 50, wherein the membrane is in contact with a membrane support, and wherein the membrane support is configured to maintain the membrane in a substantially planar orientation during use.

83. The method of claim 50, further comprising:

passing a background fluid across the porous membrane;

detecting an image of matter captured on the porous membrane after passing the background fluid through the porous membrane; and

cleaning the surface of the porous membrane;

comparing the image of matter captured on the porous membrane after passing the fluid containing one or more analytes through the membrane to the image of matter captured on the porous membrane after passing the background fluid through the porous membrane.

84. The method of claim 50, wherein the images are collected using a detector, and wherein a programmable controller is coupled to the detector.

85. The method of claim 50, further comprising applying a stain to the fluid.

86. The method of claim 50, wherein the visualization agent comprises a stain, wherein the stain is configured to emit light only in a specified portion of the visible spectrum.

87. The method of claim 50, wherein the first mask is a binary mask.

88. The method of claim 50, wherein the second mask is a binary mask.

89. The method of claim 50, wherein the visualization agent comprises a stain, and wherein further the stain is configured to emit light only in a green portion of the visible spectrum, and wherein the second wavelength of light comprises a blue portion of the visible spectrum, and wherein the third wavelength of light comprises a red portion of the visible spectrum, and wherein subtracting the first mask and the second mask from the image of the collected material in white light comprises isolating the matter on the membrane that only emits light in the green portion of the visible spectrum.

94. The method of claim 50, wherein the wavelengths of light are selected from a plurality of visible light wavelengths.

55-68. -68. (canceled)

69. A method of sensing an analyte in a fluid comprising:

passing the fluid across a porous membrane configured to capture the analyte on the porous membrane;

applying a visualization agent to the particles on the porous membrane;

detecting an image of matter captured on the porous membrane with a detector at a plurality of wavelengths of light;

detecting an image of matter captured on the porous membrane at a specific wavelength of light, wherein the specific wavelength of light represents light that is not indicative of the presence of the analyte.

90. The method of claim 69, wherein the images are collected using a CCD detector.

91. The method of claim 69, wherein the images are collected using a CCD detector coupled to a microscope.

92. The method of claim 69, wherein the images are collected using a detector, and wherein a programmable controller is coupled to the detector.

93. The method of claim 69, wherein the visualization agent comprises a stain, wherein the stain is configured to emit light only in a specified portion of the visible spectrum.

70-80. -80. (canceled)