HomeIndo-1
Indo-1 is the ratiometric dye of choice for flow cytometry although in the past the requirement for excitation by a UV source has limited it's use to cell sorters this has changed recently with the availability of small air cooled UV sources.
Indo-1 functions by changing spectral properties, when bound to calcium the dye emits at 390 nm (violet) and when unbound emits at 500 nm (green).
The colour change that Indo-1 undergoes from green to violet as the cells are activated and the calcium concentration increases in the cytoplasm and the dye binds more calcium can be plotted dynamically by programming the flow cytometer to display the two signals as a ratio against time is shown in the next figure.
An advantage of using Indo-1 as opposed to Fluo-4/Fura RED as a ratiometric calcium dye is that the Argon laser can be used solely for immunophenotyping mixed cell populations like lymphocytes and dendritic cells.
- Calcium ionophore activation of T-cells
- Effect of external calcium on Indo-1 calcium flux
- DCs can be loaded with Indo-1 at 3 uM immunophenotyped with CD8a-FITC, CD11c-PE and CD4-Cychrome and then activated by Ionomycin
- Protocol
Publications
P Dua, G Warnes, A Chruch, D Martino, D Baker, G Giovannoni, UC Meier. Functional effects of anti-neuronal antibodies from patients with Tourette's syndrome and disorders associated with streptococcal infection. Neurology Psychiatry and Brain Research; 20(1):17–18 2014.
