Adaptive control of two-photon fluorescence from green fluorescent protein by shaped femtosecond excitation pulses

2003 ◽  
Author(s):  
Hiroyuki Kawano ◽  
Yasuo Nabekawa ◽  
Akira Suda ◽  
Yu Oishi ◽  
Hideaki Mizuno ◽  
...  
Keyword(s):  
Adaptive Control ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Two Photon ◽  
Green Fluorescent ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

Automatic Counting of Intra-Cellular Ribonucleo-Protein Aggregates in Saccharomyces cerevisiae Using a Textural Approach

2019 ◽  
Vol 25 (1) ◽  
pp. 164-179
Author(s):  
Ambroise Marin ◽  
Emmanuel Denimal ◽  
Lucie Bertheau ◽  
Stéphane Guyot ◽  
Ludovic Journaux ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fluorescent Protein ◽  
Protein Aggregates ◽  
Point Of View ◽  
Zernike Moment ◽  
Two Photon ◽  
Haralick Features ◽  
Green Fluorescent ◽  
Two Photon Fluorescence

AbstractIn the context of microbiology, recent studies show the importance of ribonucleo-protein aggregates (RNPs) for the understanding of mechanisms involved in cell responses to specific environmental conditions. The assembly and disassembly of aggregates is a dynamic process, the characterization of the stage of their evolution can be performed by the evaluation of their number. The aim of this study is to propose a method to automatically determine the count of RNPs. We show that the determination of a precise count is an issue by itself and hence, we propose three textural approaches: a classical point of view using Haralick features, a frequency point of view with generalized Fourier descriptors, and a structural point of view with Zernike moment descriptors (ZMD). These parameters are then used as inputs for a supervised classification in order to determine the most relevant. An experiment using a specific Saccharomyces cerevisiae strain presenting a fusion between a protein found in RNPs (PAB1) and the green fluorescent protein was performed to benchmark this approach. The fluorescence was observed with two-photon fluorescence microscopy. Results show that the textural approach, by mixing ZMD with Haralick features, allows for the characterization of the number of RNPs.

scholarly journals Illuminating the Formation of Lumens

2007 ◽  
Vol 15 (3) ◽  
pp. 3-5
Author(s):  
Stephen W. Carmichael
Keyword(s):  
Animal Model ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Solid Core ◽  
Red Fluorescent Protein ◽  
Neuronal Process ◽  
Two Photon ◽  
Green Fluorescent

How do lumens form? Two mechanisms that come readily to mind are a wrapping model, similar to the wrapping of the myelin sheath around a neuronal process, and a solid core of cells followed by apoptosis of the central cells. Another obvious mechanism that was suggested over 100 years ago is the fusion of intracellular vacuoles. Whereas several recent studies have supported this latter mechanism, it has not yet been proven. Now, the appropriate animal model (zebrafish), the modern techniques (transgenic chimeras), dyes (green fluorescent protein and monomeric red fluorescent protein) that can be linked to proteins to label vacuoles, and two-photon imaging in real time finally have provided the strongest support yet. In an article by Makoto Kamei, Brian Saunders, Kayla Bayless, Louis Dye, George Davis, and Brant Weinstein the assembly of endothelial tubes from intracellular vacuoles was observed in vitro and in vivo.

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