Quantitative Fluorescence Imaging with Laser Scanning Confocal Microscopy

Scanning confocal microscopy (SCM) offers a dramatic instrumental advantage for fluorescence microscopy through discrimination against out-of-focus background fluorescence, through inherent resolution perpendicular to the plane of focus and improved in-plane resolution. Quantitative measurements of fluorescence intensity in such images can provide precise image determinations of fluorescence marker distributions in three dimensions. This paper aims to define the realizable and ultimately limiting capabilities of SCM for quantitative fluorescence imaging.

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