Protocols in Current Issue
    A Microfluidic Platform for Tracking Individual Cell Dynamics during an Unperturbed Nutrients Exhaustion
    Authors:  Théo Aspert, Basile Jacquel and Gilles Charvin, date: 07/20/2022, view: 1132, Q&A: 0

    Microorganisms have evolved adaptive strategies to respond to the autonomous degradation of their environment. Indeed, a growing culture progressively exhausts nutrients from its media and modifies its composition. Yet, how single cells react to these modifications remains difficult to study since it requires population-scale growth experiments to

    Plasmodium cynomolgi Berok Growth Inhibition Assay by Thiol-reactive Probe Based Flow Cytometric Measurement
    Authors:  Jessica Jie Ying Ong, Bruce Russell and Jin-Hee Han, date: 09/05/2021, view: 1841, Q&A: 0

    The relapsing malaria species, Plasmodium vivax, is the most widely distributed and difficult-to-treat cause of human malaria. The merozoites of P. vivax preferentially invade ephemeral human CD71+ reticulocytes (nascent reticulocytes), thereby limiting the development of a robust continuous culture in vitro. Fortunately, P. vivax’s sister

    Yeast Lipid Extraction and Analysis by HPTLC
    Authors:  Dan Li, Zheng-Tan Zhang, Cheng-Wen He and Zhiping Xie, date: 07/05/2021, view: 2507, Q&A: 0

    The diversity of lipid structures, properties, and combinations in biological tissues makes their extraction and analysis an experimental challenge. Accordingly, even for one of the simplest single-cellular fungi, the budding yeast (Saccharomyces cerevisiae), numerous extraction and analysis protocols have been developed to separate and quantitate

    Sex-specific Separation of Plasmodium falciparum Gametocyte Populations
    Authors:  Melanie C. Ridgway, Daniela Cihalova and Alexander G. Maier, date: 06/05/2021, view: 2257, Q&A: 0

    Plasmodium falciparum is a unicellular eukaryotic parasite that causes malaria in humans. The parasite is spread by Anopheles mosquitoes after ingestion of sexual stage parasites known as gametocytes. Malaria transmission depends on parasites switching from the disease-causing asexual blood forms to male and female gametocytes. The current

    Isolation of GFP-expressing Malarial Hypnozoites by Flow Cytometry Cell Sorting
    Authors:  Annemarie Voorberg-van der Wel, Sam O. Hoffman and Clemens H. M. Kocken, date: 05/05/2021, view: 2894, Q&A: 0

    Hypnozoites are dormant liver-stage parasites unique to relapsing malarial species, including the important human pathogen Plasmodium vivax, and pose a barrier to the elimination of malaria. Little is known regarding the biology of these stages, largely due to their inaccessible location. Hypnozoites can be cultured in vitro but these cultures

    Purification of Rice Stripe Virus
    Authors:  Gang Lu, Min Yao, Yijun Zhou and Xiaorong Tao, date: 03/20/2020, view: 2755, Q&A: 0
    [Abstract] Although many spherical and rod-shaped plant virus purification protocols are now available, only a few protocols on filamentous plant virus purification have been published. Here, we report a protocol for large-scale purification of Rice stripe virus (RSV) from RSV-infected rice tissues. RSV virions with high infectivity were first precipitated ...
    Experimental Setup for a Diffusion Bioreactor to Isolate Unculturable Soil Bacteria
    Authors:  Dhiraj Kumar Chaudhary and Jaisoo Kim, date: 10/05/2019, view: 4381, Q&A: 0
    [Abstract] Unculturable bacteria are those bacteria which proliferate in their native habitat but unable to grow or thrive in the normal laboratory media and conditions. The molecular techniques have revealed the significance of these uncultured bacteria in terms of their functional diversity and potential to produce secondary metabolites. To achieve these ...
    Yeast Single-cell RNA-seq, Cell by Cell and Step by Step
    [Abstract] Single-cell RNA-seq (scRNA-seq) has become an established method for uncovering the intrinsic complexity within populations. Even within seemingly homogenous populations of isogenic yeast cells, there is a high degree of heterogeneity that originates from a compact and pervasively transcribed genome. Research with microorganisms such as yeast ...
    Detachment Procedure of Bacteria from Atmospheric Particles for Flow-cytometry Counting
    Authors:  Carolina M. Araya, Alberto Cazorla and Isabel Reche, date: 06/20/2019, view: 3997, Q&A: 0
    [Abstract] The protocol separates bacteria from atmospheric particles, obtaining with greater precision their abundance in the atmospheric deposition. This procedure is similar to the one used to separate bacteria in streambed sediments. The detachment procedure consists of a chemical treatment with sodium pyrophosphate and Tween 20 and a physical treatment ...
    Long-term in vitro Culture of Cryptosporidium parvum
    Authors:  Nigel Yarlett and Mary Morada, date: 08/05/2018, view: 6961, Q&A: 0
    [Abstract] Continuous in vitro growth of Cryptosporidium parvum has proved difficult and conventional in vitro culture techniques result in short-term (2-5 days) growth of the parasite resulting in thin-walled oocysts that fail to propagate using in vitro cultures, and do not produce an active infection using ...

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