Photobleaching Correction in Fluorescence Correlation Spectroscopy

Fluorescence photobleaching correction for expectation-maximization algorithm

In three-dimensional (3-D) fluorescence microscopy, a series of two-dimensional (2-D) images is collected at different focal settings through the specimen. Each image in this series contains the in-focus plane plus contributions from out-of-focus structures that blur the image. Furthermore, as the series is collected the fluorescent dye in the specimen fades over time in response to the total e...

Measuring, in solution, multiple-fluorophore labeling by combining fluorescence correlation spectroscopy and photobleaching.

Determining the number of fluorescent entities that are coupled to a given molecule (DNA, protein, etc.) is a key point of numerous biological studies, especially those based on a single molecule approach. Reliable methods are important, in this context, not only to characterize the labeling process but also to quantify interactions, for instance within molecular complexes. We combined fluoresc...

Fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy.

This article focuses on methods based on fluctuation correlation spectroscopy to determine the formation of protein complexes in living cells. We present the principles of the fluctuation method applied to cells. We discuss the novelty and the promises of this approach. The emphasis is in the discussion of the underlying statistical assumptions of the image correlation spectroscopy analysis rat...

Monitoring of photobleaching in photodynamic therapy using fluorescence spectroscopy.

Photodynamic therapy (PDT) utilizing aminolevulinic acid (ALA) as a photosensitiser has been used to ablate premalignant/malignant skin conditions including superficial basal cell carcinoma (BCC) with acceptable cosmetic outcome. Clinicians continue however to face difficulties in determining the exact dose that is sufficient to achieve a complete healing from the condition where the experience...

Fluorescence correlation spectroscopy.