Table 1 One- and two-photon photophysical parameters of eLACCO2.1

From: Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging

l-Lactate

eLACCO2.1

eLACCO1.1 (ref. 9)

iLACCO1 (ref. 19)

jGCaMP8s (ref. 48)

EGFP

+

+

+

a

+a

Relative fraction of neutral chromophore (ρN)

0.97

0.24

0.97

0.57

0.97

0.79

0.74b

0.17b

0.07c

Relative fraction of anionic chromophore (ρA)

0.03

0.76

N/A

0.43

N/A

0.21

0.26b

0.83b

0.93c

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

42

45

34

38

39

39

N/A

N/A

N/A

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

76

65

78

89

81

74

2.1

57

56.0d; 58.3e

Neutral quantum yield (φN)

0.06

N/A

0.20

0.23

N/A

N/A

N/A

N/A

N/A

Anionic quantum yield (φA)

0.47

0.82

0.60

0.78

0.72

0.84

0.47

0.53

0.67d; 0.76e

Molecular brightness (ρA × εA × φA)

1.1

41

4.5f

30

0.74

13

0.26

25

34.9d; 41.2e

Neutral one-photon absorbance peak (nm)

395

398

398

397

400

400

N/A

N/A

N/A

Anionic one-photon absorbance peak (nm)

493

494

493

496

492

492

N/A

N/A

488d

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

N/A

32 (932)

N/A

48 (924)

N/A

42 (928)

N/A

25

54 (911)e

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

0.88h (932)

20 (932)

2.2 (924)

16 (924)

0.52 (928)

7.3 (928)

N/A

N/A

41 (911)e

  1. N/A not applicable.
  2. aCa2+.
  3. bCalculated from an equation, log10([anionic]/[neutral]) = pH - pKa, where pH is 7.2 and pKa is 7.65 (Ca2+ free) and 6.51 (Ca2+ bound).
  4. cCalculated from an equation, log10([anionic]/[neutral]) = pH - pKa, where pH is 7.2 and pKa is 6.1 (ref. 49).
  5. dData from ref. 49.
  6. eData from ref. 50.
  7. fEstimated from the independently measured F1,bound/F1,free ratio at 500 nm.
  8. gTwo-photon brightness is calculated as F2 = ρA × s2,A × φA at the peak wavelength (presented in parentheses), where s2,A is the peak two-photon absorption cross section.
  9. hEstimated from the independently measured F2,bound/F2,free ratio at 932 nm.