MetaFluor Acquisition

Powerful Real Time Processing Acquisition

When acquiring from video sources, MetaFluor can average up to 256 images per time point, significantly reducing random image noise. Background subtraction is also used to improve accuracy by correcting for stray light, camera noise and auto-fluorescence. Real Time Processing MetaFluor will perform frame integration or averaging and background subtraction on your image as your experiment progresses. Ratio shifts or ion fluxes are observed immediately, providing instant feedback on your experiment.

Pseudocolor vs. Intensity Modulated Display mode. The traditional pseudocolor (left) loses image definition of cells and cell structures. The same image data, displayed with the IMD representation (right), pulls cells out from the background without the need for thresholding.

Calibration

A direct display of intracellular ion concentrations is obtained by using the various calibration options offered; the Grynkiewicz equation (Grynkiewicz et al., 1985) and titration equation for both in situ and in vitro experiments. These calibrations can then be stored for future use.

G. Grynkiewicz, M. Poenie, R.Y. Tsien. A new generation of Ca2+ indicators with greatly improved fluorescence properties. The Journal of Biological Chemistry. 260(6):3440-3450 (1985).

Binning and Super Pixels

If the signal is too small or noisy, binning will improve the signal-to-noise (S/N) ratio. Binning groups of neighboring pixels creates a "super pixel" with an intensity equal to the sum of the intensity values of the original pixels. Spatial resolution is diminished, but the S/N is enhanced. If your goal is to improve the precision of ion concentration measurements and detect small changes, binning offers a powerful solution.