Table 2 One- and two-photon photophysical parameters of R-iLACCO1

From: Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging

l-Lactate

R-iLACCO1

R-eLACCO2 (ref. 18)

R-GECO1 (ref. 35)

mApple (ref. 49)

+

+

a

+a

Relative fraction of neutral chromophore (ρN)

0.95

0.39

0.88

0.35

0.94b

0.18b

0.17c

Relative fraction of anionic chromophore (ρA)

N/A

0.61

0.12

0.65

0.06b

0.82b

0.83c

Neutral extinction coefficient (mM−1 cm−1, εN)

36

55

29

N/A

33

30

N/A

Anionic extinction coefficient (mM−1 cm−1, εA)

84

88

13

59

89

69

75

Neutral quantum yield (φN)

N/A

N/A

N/A

N/A

N/A

N/A

N/A

Anionic quantum yield (φA)

0.17

0.21

0.11

0.28

0.15

0.21

0.46

Molecular brightness (ρA × εA × φA)

N/A

11.3

0.17

10.7

0.80

12

28.6

Neutral one-photon absorbance peak (nm)

444

447

444

ND

N/A

N/A

N/A

Anionic one-photon absorbance peak (nm)

579

561

578

567

577

563

N/A

Two-photon cross section (GM) (position/nm)

28 (1080)

72 (1048)

35 (1080) 236 (740)

51 (1060) 125 (740)

24 (1072)

31 (1056)

N/A

Two-photon brightness F2 (GM) (position/nm)

N/A

9.2 (1048)

0.46 (1080) 3.1 (740)

9.3 (1060) 23 (740)

0.21 (1072)

5 (1056)

N/A

  1. N/A not applicable, ND not determined.
  2. aCa2+.
  3. bCalculated from an equation, log10([anionic]/[neutral]) = pH - pKa, where pH is 7.2 and pKa is 7.69 (Ca2+ free) and 6.36 (Ca2+ bound).
  4. cCalculated from an equation, log10([anionic]/[neutral]) = pH - pKa, where pH is 7.2 and pKa is 6.5.