Applications

1.Exact oxygen measurement in cell cultures or tissue

2.Ideal for in vitro hypoxia conditions, like 3D cultures, spheroid models, and tissue

In addition to controlling the oxygen concentration in the stage top incubator, it is indispensable to know the real oxygen concentration near the cells (monitoring extracellular O₂), or even inside the cells (intracellular O₂). Because cells consume oxygen, the concentrations are typically much lower in cell clusters, such as tissue or spheroids.

With the ibidi OPAL Optical O₂ Measurement Sytem, the oxygen concentration can be measured optically and directly in a 2D cell culture dish, a compact 3D cell spheroid, or even in a piece of tissue. Oxygen-sensitive beads or cell-permeable fluorophores are utilized to measure the fluorescence lifetime.

Note: The OPAL System measures lifetimes in the range of 1-1,000 µsec (i.e., phosphorescence). To keep things simple, ibidi uses the more common term fluorescence, instead of phosphorescence.

Working Principle and Components of the ibidi OPAL System

The ibidi OPAL System is easily connected to your fluorescence microscope.

Compatibility to Microscopes

The ibidi OPAL System can be connected to all modern inverted microscopes from Nikon, Olympus, Leica, and Zeiss. Small adaptations need to be made, and during this process, ibidi will comprehensively support you. These adaptations include a specialized OPAL Filter Set and an LED Light Source Coupler for the microscope (included in the ibidi OPAL System). The OPAL Detector Unit fits the standard C-Mount camera port, or can also be mounted on an ocular.

The selected objective lens defines the measured area. An iris is used to reduce the field of measurement. The density of the CPOx beads or NanO₂ labeled cells can be selected to further specify the sample area.

Extracellular O₂ Monitoring

Oxygen monitoring is defined as the measurements of O₂ in the direct neighborhood of cultured cells. Oxygen sensitive beads, with a diameter of 50 µm (CPOx), are used in combination with the ibidi OPAL system or a FLIM microscope to measure the oxygen concentration.

Intracellular O₂ Measurement

For intracellular oxygen measurements, an oxygen-sensitive nanoparticle reagent is brought into the cells. The ibidi OPAL System or FLIM microscopes reveal quantitative O₂concentrations by identifying a change in the lifetime of the fluorophore NanO₂.

Overview: O₂ Monitoring and O₂ Imaging