Search results for "Fluorescence microscope"

E-beam crosslinked nanogels conjugated with monoclonal antibodies in targeting strategies

2017

Abstract Poly(N-vinyl pyrrolidone)-based-nanogels (NGs), produced by e-beam irradiation, are conjugated with monoclonal antibodies (mAb) for active targeting purposes. The uptake of immuno-functionalized nanogels is tested in an endothelial cell line, ECV304, using confocal and epifluorescence microscopy. Intracellular localization studies reveal a faster uptake of the immuno-nanogel conjugate with respect to the ‘bare’ nanogel. The specific internalization pathway of these immuno-nanogels is clarified by selective endocytosis inhibition experiments, flow cytometry and confocal microscopy. Active targeting ability is also verified by conjugating a monoclonal antibody which recognizes the αv…

Imaging Noncanonical Autophagy and LC3-Associated Phagocytosis in Cultured Cells

2019

International audience; Monitoring of ATG8 proteins by western blotting and immunofluorescence microscopy are the most common methods to monitor the autophagy pathway. However, it has recently been shown that ATG8 proteins can be lipidated to non-autophagosome, single-membrane compartments through a noncanonical autophagy pathway. This is commonly found to occur during macro-endocytic processes such as phagocytosis, where it has been termed LC3-associated phagocytosis, and upon lysosomotropic drug treatment. Therefore, care is required when interpreting data based on ATG8 in order to conclude whether a signal relates to the canonical or noncanonical pathway. Here we provide methods to monit…

Swift light sheet volumetric charting of large human brain portions

2020

Using a custom light sheet fluorescence microscope, we image large stained human brain portions, labelled for NeuN and GAD67 neuronal markers, discerning the inhibitory population via neural-network based image analysis and exposing the brain connectivity.

Fluorescence Microscopy as a Diagnostic Tool for Dermatophytosis.

2016

Dermatophytosis is a superficial fungal infection of keratinized tissues. Dermatophytes can cause discomfort but are not usually life threatening. However, the infection can spread and may lead to systemic fungal infections in immunocompromised patients. Currently available diagnostic methods include potassium hydroxide (KOH) testing and periodic acid-Schiff (PAS) staining. However, most diagnostic techniques cannot be performed rapidly; days to weeks may be required for conclusive results. Certain dermatophytes autofluoresce and can be observed under fluorescence microscopy. The authors examined a series of 24 cases of hematoxylin and eosin-stained dermatophytoses using fluorescence micros…

Preferential localization of Lactococcus lactis cells entrapped in a caseinate/alginate phase separated system.

2013

International audience; This study aimed to entrap bioprotective lactic acid bacteria in a sodium caseinate/sodium alginate aqueous two-phase system. Phase diagram at pH = 7 showed that sodium alginate and sodium caseinate were not miscible when their concentrations exceeded 1% (w/w) and 6% (w/w), respectively. The stability of the caseinate/alginate two-phase system was also checked at pH values of 6.0 and 5.5. Lactococcus lactis subsp. lactis LAB3 cells were added in a 4% (w/w) caseinate/1.5% (w/w) alginate two-phase system at pH = 7. Fluorescence microscopy allowed to observe that the caseinate-rich phase formed droplets dispersed in a continuous alginate-rich phase. The distribution of …

Cholesterol facilitates interactions between α‐synuclein oligomers and charge‐neutral membranes

2015

AbstractOligomeric species formed during α-synuclein fibrillation are suggested to be membrane-disrupting agents, and have been associated with cytotoxicity in Parkinson’s disease. The majority of studies, however, have revealed that the effect of α-synuclein oligomers is only noticeable on systems composed of anionic lipids, while the more physiologically relevant zwitterionic lipids remain intact. We present experimental evidence for significant morphological changes in zwitterionic membranes containing cholesterol, induced by α-synuclein oligomers. Depending on the lipid composition, model membranes are either unperturbed, disrupt, or undergo dramatic morphological changes and segregate …

Ellipsometric and fluorescence microscopic investigations of a cyclam derivative at the air/water interface

1993

Flow Cytometry versus Fluorescence Microscopy

1996

Since the pioneer work of the botanist Matthias Jakob Schleiden and the zoologist Theodor Schwann in 1839, and of Rudolph Virchow in 1859, cell research progressed in two opposite directions. Cell biologists focused their increasingly more powerful microscopies into the cell structure to reveal the great morphological complexity of the cytoplasm. A growing number of subcellular organelles thus challenged the early biochemists to discover their specific molecular features and their coordination to maintain an ordered cell life. The biochemists’ answer to such a challenge consisted usually in tearing apart cells into their discrete components and obtaining information on molecules and pathway…

Osseous response on linear and cyclic RGD-peptides immobilized on titanium surfaces in vitro and in vivo

2017

Biomimetic surface modifications of titanium implants using the Arg-Gly-Asp-sequence (RGD) are promising to accelerate bone healing in cases of medical implants. Therefore, we compared the impact of linear and cyclic RGD (l- and c-RGD) covalently coupled onto titanium surfaces on the osseous response in vitro and in vivo. In vitro, osteoblasts' behaviour on different surfaces (unmodified, amino-silanized (APTES), l- and c-RGD) was analysed regarding adhesion (fluorescence microscopy), proliferation (resazurin stain) and differentiation (RT-PCR on alkaline phosphatase (AP) & osteocalcin (OC)). In vivo, osteosynthesis screws (unmodified n=8, l-RGD n=8, c-RGD n=8) were inserted into the proxim…

BODIPY-phosphane as a versatile tool for easy access to new metal-based theranostics

2012

A new BODIPY-phosphane was synthesized and proved to be a versatile tool for imaging organometallic complexes. It also led to easy access to a new family of theranostics, featuring gold, ruthenium and osmium complexes. The compounds' cytotoxicity was tested on cancer cells, and their cell uptake was followed by fluorescence microscopy in vitro.