Methods and Algorithms for cell enumeration in low-cost cytometer

F.A.W. Coumans (Inventor), Jan Greve (Inventor), Frank P. Modica (Inventor), Leonardus Wendelinus Mathias Marie Terstappen (Inventor), Arjan G.J. Tibbe (Inventor), John A. Verrant (Inventor)

Research output: PatentProfessional

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Abstract

The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.
Original languageEnglish
Patent numberUS 7,764,821 B2
Priority date27/07/10
Publication statusSubmitted - 27 Jul 2010

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Fluids
Costs
Flow cytometry
CCD cameras
Image analysis
Magnets
Light emitting diodes
Contamination
Monitoring

Keywords

  • METIS-274650
  • IR-77361

Cite this

Coumans, F. A. W., Greve, J., Modica, F. P., Terstappen, L. W. M. M., Tibbe, A. G. J., & Verrant, J. A. (2010). Methods and Algorithms for cell enumeration in low-cost cytometer. Manuscript submitted for publication. (Patent No. US 7,764,821 B2).
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abstract = "The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.",
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Coumans, FAW, Greve, J, Modica, FP, Terstappen, LWMM, Tibbe, AGJ & Verrant, JA 2010, Methods and Algorithms for cell enumeration in low-cost cytometer, Patent No. US 7,764,821 B2.

Methods and Algorithms for cell enumeration in low-cost cytometer. / Coumans, F.A.W. (Inventor); Greve, Jan (Inventor); Modica, Frank P. (Inventor); Terstappen, Leonardus Wendelinus Mathias Marie (Inventor); Tibbe, Arjan G.J. (Inventor); Verrant, John A. (Inventor).

Patent No.: US 7,764,821 B2.

Research output: PatentProfessional

TY - PAT

T1 - Methods and Algorithms for cell enumeration in low-cost cytometer

AU - Coumans, F.A.W.

AU - Greve, Jan

AU - Modica, Frank P.

AU - Terstappen, Leonardus Wendelinus Mathias Marie

AU - Tibbe, Arjan G.J.

AU - Verrant, John A.

PY - 2010/7/27

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N2 - The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.

AB - The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.

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Coumans FAW, Greve J, Modica FP, Terstappen LWMM, Tibbe AGJ, Verrant JA, inventors. Methods and Algorithms for cell enumeration in low-cost cytometer. US 7,764,821 B2. 2010 Jul 27.