Quantitative analysis of fluorescent image – from descriptive to computational microscopy

While cellular analysis requires a great deal of caution, descriptive tissue analysis is even more subjective and interpretations are highly dependent on personal experience of the observer. Baak, in his article [Baak 2002] addressed difficulties in qualitative assessment of microscopic samples in pathology. The reliability was acceptable when an observation was assessed by at least three specialists, but consistency was low especially in more complex analysis.

Comparison of the data between different laboratories is also difficult considering the simplicity of observations made and the complexity of the pathological processes. Modern imaging methods which use charged-coupled devices (CCDs) or photomultipliers (PMTs) provide a possibility to capture the visual observations in a numerical, digitalized form. In this format reliable quantitative data can be extracted which open new ways of understanding and supporting an observation process.

In this chapter, various methods of quantitative analysis will be presented and discussed, applicable to studies of a single cell as well as whole populations and tissues.