LSM 710 and LSM 780
LEFT TOOL AREA AND HARDWARE CONTROL TOOLS
Systems
Processing Tab
Carl Zeiss
02/2010 M60-1-0025
e
129
Ratiometric Dyes - Equation Calibration
(Grynkiewicz)
(Fig. 137)
Fura-2, Indo-1,..
K
D
(dissociation constant) taken from literature
R
min
: derived from ion-free state of the dye (e.g. 0
Ca
2+
)
Rmax: derived from ion-bound state of the dye
(e.g. saturated with Ca
2+
)
F
min2
and F
max2
are the minimum and maximum
fluorescence intensities at wavelength 2
R
min
, R
max
, F
min2
and F
max2
may be determined in
the cells under investigation (in situ) or in solutions
(in vitro)
Calibration parameters may be saved and reloaded
(*.cal)
Options for Calibration Image Selection (Equation- or Titration Calibration)
•
Click into image window.
•
Select source channel(s).
•
Optional background subtraction
•
Optional calculation of parameters from overlay region(s)
Fig. 137
Ion Concentration - Ratiometric dye,
Equation Calibration
