Ratiometric Dyes - Zeiss LSM 880 Operating Manual

ZEISS
5.4.9.2

Ratiometric Dyes

• Fura-2, Indo-, SNARF, Cameleon, Ratiometric Pericam, Phluorin.
• Display fluorescence ratio R over time
• Display fluorescence ratio R corrected for background/autofluorescence over time
• Calculate absolute ion concentrations (pixel by pixel) via titration calibration (known ion concentrations
applied to the cells – in situ – or in solutions – in vitro or equation calibration where possible [Fura-2,
Indo-, SNARF]
• Calculation of R eliminates artifacts and uncertainties caused by
− inhomogeneous dye distribution
− photobleaching
− may be applied with moving cells.
Ratiometric Dyes - Online Ratio (Fig. 418)
R(t1) = F1(t1) / F2(t1), R(t2) = F1(t2) / F2(t2) .
Fig. 418
316
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
Ratiometric Dyes – online ratio
000000-2071-464
LSM 880
10/2014 V_01