MetaFluor Applications

 

	MDCK cells loaded with fura-2 AM. Imaged at the AQLM course in Woods Hole, MA. Upper right and left quadrant are 340nm and 380nm excitation images; lower left is a ratio map using IMD display. Lower right is a graph of selected cell ratio changes measured over time.

MetaFluor is an ideal tool for:

• Ratio imaging
• Calcium imaging
• FRET
• pH measurements
• Ion concentration
• Intensity-over-time

MetaFluor provides the flexibility to measure Fluorescence Resonance Energy Transfer (FRET). FRET involves the non-radiative transfer of energy from a fluorophore in an excited state to a nearby acceptor fluorophore. FRET will occur when fluorophores are within angstroms of one another. This technique is used to infer protein-protein interaction and colocalization.

Ratio Imaging

Once acquired, the wavelengths are grouped into two pairs of ratiometric wavelengths, and one isosbestic or transmitted-light image. With this arrangement, it is possible to monitor two indicators simultaneously, such as BCECF and Fura-2 for pH and calcium respectively, while also obtaining a brightfield image of cellular morphology.

Simultaneous Emission-Splitting

MetaFluor supports multi-wavelength emission-splitter acquisition. The Dual-View™ device option separates the fluorescent image into a set of two or four spectrally-discrete images and acquires them on a single CCD chip with a single exposure without overlap.

Using the TwinCam option, the Dual-Cam™ multi-wavelength emission splitter device is used to project one wavelength to one camera and a different wavelength to a second camera, allowing simultaneous acquisition from two cameras. This allows the measurement of emission-shifted probes (Indo-1, SNARF, JC-1) or FRET-based sensors (CFP, YFP) at very high speeds, without any moving parts.