Fluorescence intensity analysis through simplex optimization in flow cytometry.
Fluorescence intensity analysis in flow cytometric surface immunophenotyping has recently been appreciated in clinical applications. A curve fitting method to estimate the mean and SD values of fluorescence intensity is described in this report. A Gaussian distribution is aimed to be adapted for a specified distribution in logarithmically scaled histogram data through the simplex optimization, one of the non-linear least squares methods. In comparison with the conventional methods which include the detection of peak point and the direct calculation, this fitting method has demonstrated exceeding precisions in the estimation of both parameters with limited involved cell counts in typical lymphocytic phenotyping. The actual estimation for a precise SD value will develop the quality control approaches based on the fluorescence intensity analysis. While this method is not suitable for distributions that involve extremely small cell counts or that deviate markedly from a symmetric Gaussian, it has additional advantages of loose requirements, namely, narrow fitting regions, ordinarily small cell counts, practical computational periods and a simple programming.
Takase
K
K
Department of Pediatrics, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Iwaki
K
K
Gunji
T
T
Yata
J
J
D000954:Antigens, Surface / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013223:Statistics as Topic
D000954:Antigens, Surface