Table 2.
Fitting parameters for time-resolved fluorescence associated anisotropy of intracellular NADH in breast normal (Hs578Bst) and cancer (Hs578T) cells using pseudo-single point measurements. The results on free and enzyme-bound NADH in solution (PBS, pH 7.4) are also shown for comparative purpose as a function of the relative concentration ratio of mMDH ([NADH] ~ 226 μM) and LDH ([NADH] ~ 183 μM).
| α1 | τ2/ns | α2 | τ2/ns | β1 | ϕ1/ns | β2 | ϕ2/ns | |
|---|---|---|---|---|---|---|---|---|
| Cellular NADH: | ||||||||
| Hs578Bst (n = 9) | 0.3(1)a | 0.4(1) | 0.7(1) | 0.7(4) | 0.42(5) | 0.3(2) | 0.36(1) | ∞ |
| Hs578T (n = 7) | 0.4(1) | 0.5(2) | 0.6(1) | 0.8(4) | 0.36(6) | 0.18(4) | 0.40(3) | ∞ |
| Free NADH: | ||||||||
| In PBS (pH7.4) | -- | -- | -- | -- | 0.29 | 0.27 | -- | -- |
| [NADH]:[mMDH] | ||||||||
| 16:1 | 0.52 | 0.43 | 0.48 | 0.80 | 0.28 | 0.14 | 0.41 | 29.5 |
| 8:1 | 0.33 | 0.43 | 0.67 | 0.80 | 0.26 | 0.12 | 0.43 | 30.0 |
| 4:1 | 0.15 | 0.43 | 0.85 | 0.80 | 0.10 | 0.03 | 0.45 | 27.9 |
| 2.6:1 | 0.08 | 0.43 | 0.93 | 0.80 | 0.05 | 0.04 | 0.46 | 26.8 |
| 2:1 | -- | -- | -- | -- | -- | -- | 0.45 | 30.2 |
| 1:1 | -- | -- | -- | -- | -- | -- | 0.44 | 30.0 |
| [NADH]:[LDH] | ||||||||
| 32:1 | 0.88 | 0.42 | 0.12 | 1.02 | 0.21 | 0.11 | 0.09 | 39.6 |
| 16:1 | 0.80 | 0.42 | 0.20 | 1.02 | 0.26 | 0.19 | 0.39 | 43.5 |
| 8:1 | 0.63 | 0.42 | 0.37 | 1.02 | 0.26 | 0.16 | 0.41 | 36.8 |
| 5:1 | 0.48 | 0.42 | 0.52 | 1.02 | 0.24 | 0.15 | 0.42 | 36.5 |
| 4:1 | 0.38 | 0.42 | 0.62 | 1.02 | 0.24 | 0.12 | 0.42 | 43.5 |
| 2:1 and 2:2 | -- | -- | -- | -- | -- | -- | 0.414 | 39.6 |
a
The numbers in parentheses represent the standard deviation of the last digit(s). The solution measurements were repeated two times and the standard deviation is within 10 % of the stated fitting parameters.