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Microgenomic Expression Profiling

nd Cy5 are selected which will place the spectrum cursors at the appropriate wavelengths (550 nm and 650 nm respectively).
Calculate the FOI of Cy3 = (OD550/ 0.15)*(324)/(OD260*40)
Calculate the FOI of Cy5 = (OD650/ 0.25)*(324)/(OD260*40)
FOI readings > 20 indicates adequately labeled probe, suitable for array hybridization.
FOI readings between 15 and 20 are lower than recommended but still use-able. The investigator should note that signal issues with the final array data might be due to the lower FOI.
FOI readings lower than 15 should be treated with caution. These samples may not be suitable for hybridization to microarrays.

13. Perform Expression Array Analysis:
Labeled RNA input will depend on the desired type of expression array platform.


Conclusion
The concerted use of several new technologies allows research investigators to perform microgenomic expression profiling from extremely limited cell mass. LCM and LC provide the means of precise cell separation from heterogeneous tissue. Advanced isolation systems and amplification protocols produce sufficient material for microarray probe labeling. Bioanalyzer systems as well as novel microspectrophotometry bring a high level of quality assurance and confidence. The combined use of these various technologies provides a micogenomic work flow from cell separation through expression analysis with unprecedented process control.


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