Zeiss LSM 880 Operating Manual page 324

ZEISS
Fig. 420 Ion concentration - Ratiometric dye,
Equation Calibration
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)
318
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
000000-2071-464
Ratiometric Dyes
(Grynkiewicz) (Fig. 420)
Fura-2, Indo-1,
K (dissociation constant) taken from literature
D
R : derived from ion-free state of the dye (e.g. 0
min
2+
Ca )
R max: derived from ion-bound state of the dye
(e.g. saturated with Ca
F and F are the minimum and maximum
min2 max2
fluorescence intensities at wavelength 2
R , R , F and F
min max min2
the cells under investigation (in situ) or in solutions
(in vitro)
Calibration parameters may be saved and reloaded
(*.cal)
LSM 880
- Equation Calibration
2+ )
may be determined in
max2
10/2014 V_01