Search results for "Confocal"
showing 10 items of 444 documents
Reduction of focus size in tightly focused linearly polarized beams
2004
The electromagnetic theory predicts that when a linearly polarized collimated field is focused by a high-angle focusing system, components perpendicular to the initial polarization are generated. The use of annular masks to reduce the area of the focal spot usually increases the magnitude of this phenomenon, known as depolarization. We present a class of masks, the three-ring masks, which are important because they narrow the central lobe of the focal intensity distribution without increasing the depolarization. This can be very useful in modern optical applications, such as confocal microscopy or multiphoton scanning microscopy.
Putative p24 complexes in Arabidopsis contain members of the delta and beta subfamilies and cycle in the early secretory pathway
2013
p24 proteins are a family of type I membrane proteins localized to compartments of the early secretory pathway and to coat protein I (COPI)- and COPII-coated vesicles. They can be classified, by sequence homology, into four subfamilies, named p24α, p24β, p24γ, and p24δ. In contrast to animals and fungi, plants contain only members of the p24β and p24δ subfamilies, the latter probably including two different subclasses. It has previously been shown that transiently expressed red fluorescent protein (RFP)-p24δ5 (p24δ1 subclass) localizes to the endoplasmic reticulum (ER) at steady state as a consequence of highly efficient COPI-based recycling from the Golgi apparatus. It is now shown that tr…
The depletion of nuclear glutathione impairs cell proliferation in 3t3 fibroblasts.
2009
BACKGROUND:Glutathione is considered essential for survival in mammalian cells and yeast but not in prokaryotic cells. The presence of a nuclear pool of glutathione has been demonstrated but its role in cellular proliferation and differentiation is still a matter of debate. PRINCIPAL FINDINGS:We have studied proliferation of 3T3 fibroblasts for a period of 5 days. Cells were treated with two well known depleting agents, diethyl maleate (DEM) and buthionine sulfoximine (BSO), and the cellular and nuclear glutathione levels were assessed by analytical and confocal microscopic techniques, respectively. Both agents decreased total cellular glutathione although depletion by BSO was more sustaine…
Functional characterization of the plastidial 3-phosphoglycerate dehydrogenase family in Arabidopsis.
2013
This work contributes to unraveling the role of the phosphorylated pathway of serine (Ser) biosynthesis in Arabidopsis (Arabidopsis thaliana) by functionally characterizing genes coding for the first enzyme of this pathway, 3-phosphoglycerate dehydrogenase (PGDH). We identified two Arabidopsis plastid-localized PGDH genes (3-PGDH and EMBRYO SAC DEVELOPMENT ARREST9 [EDA9]) with a high percentage of amino acid identity with a previously identified PGDH. All three genes displayed a different expression pattern indicating that they are not functionally redundant. pgdh and 3-pgdh mutants presented no drastic visual phenotypes, but eda9 displayed delayed embryo development, leading to aborted emb…
Modification of Plasma Membrane Organization in Tobacco Cells Elicited by Cryptogein
2014
Abstract Lipid mixtures within artificial membranes undergo a separation into liquid-disordered and liquid-ordered phases. However, the existence of this segregation into microscopic liquid-ordered phases has been difficult to prove in living cells, and the precise organization of the plasma membrane into such phases has not been elucidated in plant cells. We developed a multispectral confocal microscopy approach to generate ratiometric images of the plasma membrane surface of Bright Yellow 2 tobacco (Nicotiana tabacum) suspension cells labeled with an environment sensitive fluorescent probe. This allowed the in vivo characterization of the global level of order of this membrane, by which w…
Lightfield microscopy, an emerging tool for real-time 3D imaging
2020
Integral, or lightfield, microscopy offers the possibility of capturing and processing in real time multiple views of 3D fluorescent samples captured with a single shot. In this contribution we review the recent advances in lightfield microscopy and enunciate the forthcoming challenges.
Fast multi-directional DSLM for confocal detection without striping artifacts
2020
In recent years light-sheet fluorescence microscopy (LSFM) has become a cornerstone technology for neuroscience, improving the quality and capabilities of 3D imaging. By selectively illuminating a single plane, it provides intrinsic optical sectioning and fast image recording, while minimizing out of focus fluorescence background, sample photo-damage and photo-bleaching. However, images acquired with LSFM are often affected by light absorption or scattering effects, leading to un-even illumination and striping artifacts. In this work we present an optical solution to this problem, via fast multi-directional illumination of the sample, based on an acousto-optical deflector (AOD). We demonstr…
Tunable axial superresolution by annular binary filters. Application to confocal microscopy
1995
We present a set of annular binary pupil filters for increasing the axial resolving capacity of imaging systems. The filters consist of two transparent annuli of the same area. It is shown that by changing the area of the transparent regions it is possible to obtain a tunable reduction of the width of the central lobe of the axial point spread function of the imaging system. However, this reduction is accompanied by a severe increase of the strength of secondary lobes, what can make these filters not very useful when used in conventional imaging systems. That is why we propose to use these filters for apodizing confocal microscopy systems. It is shown that in this case an important reductio…
Sidelobe decline in single-photon 4Pi microscopy by Toraldo rings.
2003
We demonstrate theoretically the feasibility of single-photon 4Pi-confocal microscopy. By inserting a pair of properly designed multi-ring phase-only pupil filters in the illumination path of a 4Pi microscope the height of the sidelobes of the point spread function substantially reduced, so that there is no ambiguity in the 3D image. Then, an axial resolution up to four times higher than that of single-photon confocal microscope can be effectively achieved.
Three-dimensional superresolution by annular binary filters
1999
We present a new family of annular binary filters for improving the three-dimensional resolving power of optical systems. The filters, whose most important feature is their simplicity, permit to achieve a significant reduction, both in the transverse and in the axial direction, of the central lobe width of the irradiance point spread function of the system. The filters can be used for applications such as optical data storage or confocal scanning microscopy.