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US 5,627,040 AGranted
Flow cytometric method for autoclustering cells
AssigneeBecton, Dickinson and Company
InventorsPierre Bierre, Ronald A. Mickaels
Life of this patent
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Abstract
This invention relates to a method for autoclustering N-dimensional datastreams. The invention has particular utility in analyzing multi-parameter data from a flow cytometer, and more particularly has utility in analyzing data from whole blood cells tagged with fluorescently labelled CD3, CD4 and CD8 monoclonal antibodies to which a known number of fluorescent microbeads has been added.
Claims17
A method for classifying a sample of particles into one or more clusters comprising the steps of:
- (a) collecting a plurality of parameters for each particle in a sample of particles;
- (b) displaying said parameters for each particle in the sample of particles on a set of two-dimensional scatterplots;
- (c) manually positioning a geometric boundary surface on the set of two-dimensional scatterplots for at least one cluster of particles so as to enclose a group of the displayed particles in the cluster, the geometric boundary surface having a user-defined, center location, shape, size, radius and orientation;
- (d) defining a vector for each particle in the sample of particles and testing if each vector is contained within the geometric boundary surface;
- (e) summing each vector contained within the geometric boundary surface to calculate a vector mean;
- (f) after a user-defined number of vectors are summed to calculate the vector mean, calculating a new center location for the geometric boundary surface using the vector mean;
- (g) moving the geometric boundary surface on the set of two-dimensional scatterplots to relocate the geometric boundary surface about the new center location;
- (h) repeating steps d)-g) until a user-defined number of vectors or all available vectors have been included in the calculation of a final center location;
- (i) moving the geometric boundary surface on the set of two-dimensional scatterplots to relocate the geometric boundary surface about the final center location;
- and
- (j) comparing all subsequently received particles with the geometric boundary surface for inclusion within or exclusion outside the cluster associated with the geometric boundary surface.
8 more elements
A method for classifying a sample of blood cells in a sample of such cells into two or more clusters comprising the steps of:
- (a) manually positioning a geometric boundary surface on a series of two dimensional surfaces in the region where at least one cluster is expected, said boundary which will enclose a group of the cells displayed as a series of scatterplots on the two dimensional surfaces, so as to define a cluster the geometric boundary surface having a user defined center location shape, size, radius, and orientation;
- (b) analyzing the cells by means of flow cytometry wherein at least two parameters of data are recorded for each cell analyzed;
- (c) displaying said parameters for each blood cell in the sample of blood cells on said scatterplots;
- (d) defining a vector for each cell analyzed and testing if each vector is contained within the geometric boundary surface;
- (e) summing each vector contained within the geometric boundary surface to calculate a vector mean;
- (f) after a user-defined number of vectors are summed to calculate the vector mean, calculating a new center location for the geometric boundary surface using vector mean;
- (g) moving the geometric boundary surface on the set of two-dimensional scatterplots to relocate the geometric boundary surface about the new center location;
- (h) repeating steps d)-g) until a user defined number of vectors or all available vectors have been included in the calculation of a final center location;
- (i) moving the geometric boundary surface on the set of two dimensional scatterplots to relocate the surface about the final center location;
- and
- (j) comparing all subsequently received blood cells with the boundary surface for inclusion within or exclusion outside the cluster associated with the geometric boundary surface.
8 more elements
Dependencies are inferred from the claim text.