Patent ID: 8825431

Claim:
A fluorescence intensity correcting method, comprising: receiving fluorescences generated from plural fluorescent dyes excited by radiating a light to a microparticle multiply-labeled with said plural fluorescent dyes having fluorescence wavelength bands overlapping one another by photodetectors which correspond to different received light wavelength bands, respectively, and whose number is larger than the number of fluorescent dyes; and approximating measured spectra obtained by collecting detected values from said plural photodetectors based on a linear sum of single-dyeing spectra obtained from a microparticle individually labeled with the fluorescent dyes, wherein the approximation of the measured spectra based on the linear sum of the single-dyeing spectra is carried out by using a least-squares method, and wherein a parameter a k (k=1 to m) at which an evaluation function expressed by following Expression gets a minimum value is obtained, thereby calculating intensities of the fluorescences generated from the fluorescent dyes, respectively: χ 2 ≡ ∑ i = 1 N ⁢ [ y i - ∑ k = 1 M ⁢ a k ⁢ X k ⁡ ( x i ) σ i ] 2 where X k (x i ) represents a detected value from the i-th photodetector in the single dyeing spectrum of the k-th fluorescent dye, y i represents a detected value from the i-th photodetector in the measured spectra, and σ i represents a reciprocal number of a weight for the measured value from the i-th photodetector.