Enhancing flow cytometry discrimination with geometric transformation
In flow cytometry, particles ( 2 ) can be distinguished between populations ( 8 ) by combining n-dimensional parameter data, which may be derived from signal data from a particle, to mathematically achieve numerical results representative of an alteration ( 48 ). An alteration may include a rotational alteration, a scaled alteration, or perhaps even a translational alteration. Alterations may enhance separation of data points which may provide real-time classification ( 49 ) of signal data corresponding to individual particles into one of at least two populations.
1. A flow cytometer comprising:
a nozzle for entraining particles in a fluid stream;
a laser focusing optics for focusing a laser beam onto an interrogation point on the fluid stream;
at least one detector for producing signal data affiliated with the individual particles in the fluid stream;
one or more processors having computer readable memory and instructions written thereon to: convert the signal data affiliated with the individual particles into n-dimensional parameter data, where n is equal to the number of parameters and where the number of parameters is at least 2, wherein at least two populations of the individual particles overlap in one of the n-dimensions; mathematically alter the n-dimensional parameter data by at least rotating the n-dimensional parameter data to increase spatial separation in the dimension having the overlap; classify the n-dimensional parameter data of each of the individual particles using the rotated n-dimensional parameter data; and discriminate between the individual particles based upon the classification.
2. The flow cytometer of claim 1 , wherein the at least one detector further comprises a first detector and a second detector.
3. The flow cytometer of claim 2 , wherein the first detector further comprises a forward detector and second detector comprises a side detector.
4. The flow cytometer of claim 1 , wherein the one or more processors having computer readable memory further comprises instructions written thereon for simultaneously processing the n-dimensional parameter data to mathematically achieve numerical results of the rotational alteration in addition to a scaled alteration and a translation alteration.
5. The flow cytometer of claim 1 , wherein the one or more processors having computer readable memory further comprises instructions written thereon for generating a graphical representation of the at least two populations of the mathematically altered n-dimensional parameter data.
6. The flow cytometer of claim 5 , wherein the graphical representation of the at least two populations of the mathematically altered n-dimensional parameter data comprises an n-parameter data plot in a Cartesian coordinate system.
7. The flow cytometer of claim 1 , wherein the particles comprise sperm and wherein at least one of the two populations comprises an X-bearing sperm population or a Y-bearing sperm population.