Quantitative Intercomparison of Transmission Electron Microscopy, Flow Cytometry, and Epifluorescence Microscopy for Nanometric Particle Analysis

2002 ◽  
Vol 304 (2) ◽  
pp. 249-256 ◽  
Author(s):  
Matthew M. Ferris ◽  
Carrie L. Stoffel ◽  
Thain T. Maurer ◽  
Kathy L. Rowlen
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Epifluorescence Microscopy ◽  
Particle Analysis ◽  
Transmission Electron

Characterization of cellular development and fatty acid accumulation in thraustochytrid Aurantiochytrium strains of Taiwan

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Natarajan Velmurugan ◽  
Yesupatham Sathishkumar ◽  
Shashanka Sonowal ◽  
Ka-Lai Pang ◽  
Yang Soo Lee
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Intracellular Lipid ◽  
Fatty Acid Accumulation ◽  
Transmission Electron ◽  
Intracellular Lipid Accumulation ◽  
High Level

Abstract Long-chain saturated and polyunsaturated fatty acids of two new thraustochytrid isolates cultured from Taiwan mangroves, Aurantiochytrium sp. IMB169 and Aurantiochytrium sp. IMB171, were characterized through their cell growth and development in relation to their intracellular lipid accumulation using transmission electron microscopy. Flow cytometry in combination with the lipophilic fluorescent dye BODIPY 505/515 was used to stain and characterize intracellular lipid bodies in the two isolates. The transmission electron microscopy and flow cytometry analyses revealed a progressive accumulation of lipid products in IMB169 and IMB171. Further, selective BODIPY stained cells were successfully separated and enriched using flow cytometry at single cell level. Among the two isolates, IMB169 was found to produce a high level of docosahexaenoic acid. The qualitative and analytical results obtained using electron microscopy and flow cytometry studies were validated by gas chromatography (GC). In addition, a quantitative baseline was established using cell growth, flow cytometry and GC analyses for developing an efficient bioprocessing methodology to selectively enrich thraustochytrids phenotypes with desirable characteristics.

scholarly journals Endlicheria bracteolata (Meisn.) Essential Oil as a Weapon Against Leishmania amazonensis: In Vitro Assay

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2525 ◽  
Author(s):  
Mariana Margatto Rottini ◽  
Ana Claudia Fernandes Amaral ◽  
José Luiz Pinto Ferreira ◽  
Edinilze Souza Coelho Oliveira ◽  
Jefferson Rocha de Andrade Silva ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Essential Oil ◽  
Cutaneous Leishmaniasis ◽  
Leishmania Amazonensis ◽  
Vitro Assay ◽  
Transmission Electron ◽  
Biological Potential ◽  
Intracellular Amastigote

The difficulties encountered and the numerous side effects present in the treatment of cutaneous leishmaniasis have encouraged the research for new compounds that can complement or replace existing treatment. The growing scientific interest in the study of plants, which are already used in folk remedies, has led our group to test Endlicheria bracteolata essential oil against Leishmania amazonensis. Several species of the Lauraceae family, or their compounds, have relevant antiprotozoal activities Therefore, the biological potential on L. amazonensis forms from the essential oil of Endlicheria bracteolata leaves was verified for the first time in that work. The antileishmanial activity was evaluated against promastigotes and intracellular amastigotes, and cytotoxicity were performed with J774.G8, which were incubated with different concentrations of E. bracteolata essential oil. Transmission electron microscopy and flow cytometry were performed with E. bracteolata essential oil IC50. Promastigote forms showed E. bracteolata essential oil IC50 of 7.945 ± 1.285 µg/mL (24 h) and 6.186 ± 1.226 µg/mL (48 h), while for intracellular amastigote forms it was 3.546 ± 1.184 µg/mL (24 h). The CC50 was 15.14 ± 0.090 µg/mL showing that E. bracteolata essential oil is less toxic to macrophages than to parasites. Transmission electron microscopy showed that E. bracteolata essential oil treatment is capable of inducing mitochondrial damage to promastigote and intracellular amastigote forms, while flow cytometry showed ΔѰm disruption in treated parasites. These results could bring about new possibilities to develop products based on E. bracteolata essential oil to treat cutaneous leishmaniasis, especially for people who cannot receive the conventional therapy.

scholarly journals The Depolarization-Evoked, Ca2+-Dependent Release of Exosomes From Mouse Cortical Nerve Endings: New Insights Into Synaptic Transmission

2021 ◽  
Vol 12 ◽  
Author(s):  
Guendalina Olivero ◽  
Francesca Cisani ◽  
Danilo Marimpietri ◽  
Daniela Di Paolo ◽  
Maria Cristina Gagliani ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Light Scattering ◽  
Biochemical Analysis ◽  
Nerve Terminals ◽  
Transmission Electron ◽  
Presynaptic Release ◽  
Presynaptic Nerve Terminals ◽  
Active Release

Whether exosomes can be actively released from presynaptic nerve terminals is a matter of debate. To address the point, mouse cortical synaptosomes were incubated under basal and depolarizing (25 mM KCl-enriched medium) conditions, and extracellular vesicles were isolated from the synaptosomal supernatants to be characterized by dynamic light scattering, transmission electron microscopy, Western blot, and flow cytometry analyses. The structural and biochemical analysis unveiled that supernatants contain vesicles that have the size and the shape of exosomes, which were immunopositive for the exosomal markers TSG101, flotillin-1, CD63, and CD9. The marker content increased upon the exposure of nerve terminals to the high-KCl stimulus, consistent with an active release of the exosomes from the depolarized synaptosomes. High KCl-induced depolarization elicits the Ca2+-dependent exocytosis of glutamate. Interestingly, the depolarization-evoked release of exosomes from cortical synaptosomes also occurred in a Ca2+-dependent fashion, since the TSG101, CD63, and CD9 contents in the exosomal fraction isolated from supernatants of depolarized synaptosomes were significantly reduced when omitting external Ca2+ ions. Differently, (±)-baclofen (10 µM), which significantly reduced the glutamate exocytosis, did not affect the amount of exosomal markers, suggesting that the GABAB-mediated mechanism does not control the exosome release. Our findings suggest that the exposure of synaptosomes to a depolarizing stimulus elicits a presynaptic release of exosomes that occurs in a Ca2+-dependent fashion. The insensitivity to the presynaptic GABAB receptors, however, leaves open the question on whether the release of exosomes could be a druggable target for new therapeutic intervention for the cure of synaptopathies.

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