Table 1 Spectroscopic properties of GCaMP3, GCaMP6s and their -Q and -T mutants.

From: Absolute measurement of cellular activities using photochromic single-fluorophore biosensors and intermittent quantification

 

GCaMP3

GCaMP3-Q

GCaMP3-T

GCaMP6s

GCaMP6s-Q

GCaMP6s-T

λabs, apo (nm)

403/503

404/503

397/495

400/502

400/510

395/493

λabs, sat (nm)

396/499

400/498

394/486

401/497

405/496

399/486

λexc, apo (nm)

397/504

401/505

387/491

398/511

393/512

392/495

λexc, sat (nm)

498

497

391/484

497

496

483

λem, apo (nm)

515

515

510

518

518

512

λem, sat (nm)

513

513

506

513

513

506

\({\epsilon }_{apo}^{\,{{{{{{\rm{app}}}}}}}}\) (103 M−1 cm−1)

2.4

6.7

3.8

8.0

5.7

5.2

\({\epsilon }_{sat}^{\,{{{{{{\rm{app}}}}}}}}\) (103 M−1 cm−1)

19.0

24.3

13.4

63.7

67.5

48.0

QY of fluorescence

0.59

0.29

0.10

0.60

0.34

0.07

Molecular brightness (vs EGFP)

0.33

0.21

0.035

1.13

0.69

0.083

pKa, sat

7.6

7.6

8.1

5.9

6.0

6.2

pKa, apo

10.10

8.4

8.7

12.7

10.2

11.8

Kd (nM) (Ca2+, fluorescence)

191

177

238

181

266

261

Kd (nM) (Ca2+, \({({{\Delta }}{{{{{{{\rm{F}}}}}}}}/{{{{{{{{\rm{F}}}}}}}}}_{0})}_{{{{{{{{\rm{hv}}}}}}}}}\))

95

123

118

47

51

63

Hill coefficient

3.1

3.1

3.3

2.1

2.1

2.3

\({({{\Delta }}{{{{F}}}}/{{{{{F}}}}}_{sat})}_{{{{{{{{\rm{Ca}}}}}}}}}\) (on)

0.90

0.71

0.89

0.98

0.97

0.92

\({({{\Delta }}{{{{F}}}}/{{{{{F}}}}}_{sat})}_{{{{{{{{\rm{Ca}}}}}}}}}\) (off)

0.89

0.47

0.55

0.97

0.74

−0.11

\({({{\Delta }}{{{{F}}}}/{{{{{F}}}}}_{0})}_{{{{{{{{\rm{hv}}}}}}}}}\) (sat)

−0.19

0.65

0.83

0.082

0.86

0.93

\({({{\Delta }}{{{{F}}}}/{{{{{F}}}}}_{0})}_{{{{{{{{\rm{hv}}}}}}}}}\) (apo)

−0.44

0.39

0.31

−0.32

0.0071

−0.010

Thermal recovery, sat (s)

110

308

2388

38

330

726

  1. In the case where two values are present (e.g. 403/503), the first and second values are associated with the neutral and anionic forms of the chromophore, respectively. The extinction coefficients are ‘apparent’ in that they are expressed with respect to the total protein concentration, rather than purely the concentration of the fluorescent state, and are reported for the absorption maxima. Spectra, pH titrations and Ca2+ titrations are available in Supplementary Figs. S1–S9.