A flow cytometric comparison of Indo-1 to fluo-3 and Fura Red excited with low power lasers for detecting Ca2+ flux

Membrane potential of CD4+ T cells is a subset specific feature that depends on the direct cell-to-cell contacts with monocytes

“…Fluorescence emission for Ca 2ϩ bound indo-1 is 405 nm, whereas Ca 2ϩ free indo-1 emits light of 475 nm. Hence [Ca 2ϩ ] i can be calculated as a ratio of mean fluorescence intensity of indo-1 emission at 405 nm/mean fluorescence intensity of indo-1 emission at 475 nm (indo-1 bound/indo-1 free fluorescence) [19]. Therefore possible error arising from different dye loading into particular samples is markedly reduced.…”

Sequential Cooperation of CD2 and CD48 in the Buildup of the Early TCR Signalosome

“…Fluo-4/Fura-red loaded cells were then analyzed by flow cytometry using a LSRII flow cytometer (BD Biosciences). Calcium mobilization was monitored within the first three minutes after TCR cross-linking with the anti-TCR mAb C305 by analyzing the fluorescence emission ratio of Fura Red and Fluo-4 as previously described (27).…”

Experimental Conditions and Mathematical Analysis of Kinetic Measurements Using Flow Cytometry – The FacsKin Method

“…Another way of explaining this method is that it is essentially a smoothing of the spiked curve that goes through all the dots in the scatter plot. There are several averaging methods that could be used: mean (used in Omann et al, 1990;LundJohansen et al, 1992;Rijkers et al, 1993;do Céu Monteiro et al, 1999;Jakubczak et al, 2006;Schepers et al, 2009), geometric mean (Bailey et al, 2006), median (Szalay et al, 2012). The mean is sensitive to outliers and is suitable for the characterization of a normal distribution.…”

“…These include partitioning the whole time-frame into intervals of the same length (used in the method described in Section 3); partitioning the whole time-frame into intervals all containing the same number of cells; having a fixed-length time-frame and shifting it through the whole measurement and calculating the average in these (overlapping) intervals (moving average, used in Rijkers et al, 1993, Bailey et al, 2006; using local regression (lowess method, published in (Cleveland, 1979), used in (Szalay et al, 2012)); using cubic splines etc.…”