×150.jpgPhagocytic and endocytic activities are essential cellular processes involved in elimination of pathogens and debris, as in internalisation of some cell nutrients. Using live cell staining, we set up a kinetic acquisation of fluorescent active phagosomes.

pHrodo® Green E. coli bioparticles conjugates are non-fluorescent outside the cell at neutral pH, but fluoresce brightly green at acidic pH such as in phagosomes. Following apparition of green fluorescence, we can assess specific phagocytic and endocytic activity over time.

Using our ImageXpress, supply with CO2 and 37°C chamber, we analysed phagoctytic and endocytic activities of several cell lines, such as mouse macrophages Raw 264.7. Kinetic acquisition can be performed up to 24 hours with fluorescence and transmitted light images.

Following video illlustrate assessment of phagocytic activity of Raw264.7 activated by LPS.

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A team from Rome University “La Sapienza”, in collaboration with the Italian Space Agency, works on a new technology to measure cells numbers directly in cell culture medium, without biological markers or internal sensors.

Un gruppo di ricercatori della Sapienzaha scoperto una tecnica innovativa che permette di misurare le cellule in tempo reale. Un risultato importante sia in ambito spaziale che sulla Terra.

Source: Le cellule in tempo reale

This new microscopy technology is a promising tool for in vivo 3D imaging of fast dynamic processes in cells and embryos. Lattice light-sheet microscopy allows fast 3D superresolution, with lower photobleaching and phototoxicity.

Here is the Editor’s summary about this article published in Science 2 weeks ago (Chen et al.) :

From single molecules to embryos in living color

Animation defines life, and the three-dimensional (3D) imaging of dynamic biological processes occurring within living specimens is essential to understand life. However, in vivo imaging, especially in 3D, involves inevitable tradeoffs of resolution, speed, and phototoxicity. Chen et al. describe a microscope that can address these concerns. They used a class of nondiffracting beams, known as 2D optical lattices, which spread the excitation energy across the entire field of view while simultaneously eliminating out-of-focus excitation. Lattice light sheets increase the speed of image acquisition and reduce phototoxicity, which expands the range of biological problems that can be investigated. The authors illustrate the power of their approach using 20 distinct biological systems ranging from single-molecule binding kinetics to cell migration and division, immunology, and embryonic development.

Zebrafish brain firing

Ever wanted to see the activity of the neurons from a almost entire brain ? 10 days ago, Vladimirov et al. described how they have used light-sheet microscopy to record the activity of nearly every neuron in the larval zebrafish brain as the animal responded to sensory stimuli.

You can see it by looking at this wonderfull video.

Light-sheet fluorescence microscopy allow to illuminate a thin slice of the sample, perpendicularly to the direction of the observation.

From Vladimirov, N. et al. Light-sheet functional imaging in behaving zebrafish. Nature Methods doi:10.1038/nmeth.3040 (27 July 2014)



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