Biological Sciences

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    Protocols in Current Issue
    Cryptococcus neoformans Virulence Assay Using a Galleria mellonella Larvae Model System
    Authors:  Piotr R. Stempinski, Daniel F. Q. Smith and Arturo Casadevall, date: 08/05/2022, view: 263, Q&A: 0
    [Abstract]

    Cryptococcus neoformans is a human pathogenic fungus that can cause pulmonary infections and meningitis in both immunocompromised and otherwise healthy individuals. Limited treatment options and a high mortality rate underlie the necessity for extensive research of the virulence of C. neoformans. Here we describe a detailed protocol for using the

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    Analysis of Caenorhabditis elegans Aging-related Neurodegeneration in Chemosensory Neurons
    Authors:  Cira Crespo and Roberto Grau, date: 07/20/2022, view: 656, Q&A: 0
    [Abstract]

    Aging and neuronal deterioration constitute important risk factors for the development of neuronal-related diseases, such as different dementia. The nematode Caenorhabditis elegans has emerged as a popular model system for studying neurodegeneration diseases, due to its complete neuronal connectivity map. DiI is a red fluorescent dye that can

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    HaloChIP-seq for Antibody-Independent Mapping of Mouse Transcription Factor Cistromes in vivo
    [Abstract]

    Chromatin immunoprecipitation (ChIP) maps, on a genome-wide scale, transcription factor binding sites, and the distribution of other chromatin-associated proteins and their modifications. As such, it provides valuable insights into mechanisms of gene regulation. However, successful ChIP experiments are dependent on the availability of a

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    Protocol to Study Spatial Subgoal Learning Using Escape Behavior in Mice
    Authors:  Philip Shamash and Tiago Branco, date: 06/20/2022, view: 927, Q&A: 0
    [Abstract]

    Rodent spatial navigation is a key model system for studying mammalian cognition and its neural mechanisms. Of particular interest is how animals memorize the structure of their environments and compute multi-step routes to a goal. Previous work on multi-step spatial reasoning has generally involved placing rodents at the start of a maze until

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    Time-off-pick Assay to Measure Caenorhabditis elegans Motility
    Authors:  Alyssa C. Walker, Rohan Bhargava, Amanda S. Brust, Ali A. Owji and Daniel M. Czyż, date: 06/20/2022, view: 1172, Q&A: 0
    [Abstract]

    Caenorhabditis elegans is a simple metazoan that is often used as a model organism to study various human ailments with impaired motility phenotypes, including protein conformational diseases. Numerous motility assays that measure neuro-muscular function have been employed using C. elegans. Here, we describe “time-off-pick" (TOP), a novel

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    Protocol for High Throughput Screening of Antibody Phage Libraries
    [Abstract]

    Phage display is a proven and widely used technology for selecting specific antibodies against desired targets. However, an immense amount of effort is required to identify and screen the desired positive clones from large and diverse combinatorial libraries. On the other hand, the selection of positive binding clones

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    Electroporation of Small Interfering RNAs into Tibialis Anterior Muscles of Mice
    Authors:  Anna Stephan, Flavia A. Graca, Liam C. Hunt and Fabio Demontis, date: 06/05/2022, view: 963, Q&A: 0
    [Abstract]

    Aging and wasting of skeletal muscle reduce organismal fitness. Regrettably, only limited interventions are currently available to address this unmet medical need. Many methods have been developed to study this condition, including the intramuscular electroporation of DNA plasmids. However, this technique requires surgery and high electrical

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    Protocol to Isolate Germinal Centers by Laser Microdissection
    Authors:  Farbod Bahreini, Markus Niebuhr, Julia Belde, Jürgen Westermann and Kathrin Kalies, date: 06/05/2022, view: 655, Q&A: 0
    [Abstract]

    During adaptive immune responses, germinal centers (GC) appear as transient microstructures, in which antigen-specific B and T cells interact with each other. Because only the antigen-activated B and T cells, such as Plasmablasts or follicular T helper (Tfh) cells, are present in GC, the in depth-analysis of GC is of great interest. To identify

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    A Modified Fluctuation Assay with a CAN1 Reporter in Yeast
    Authors:  Pengyao Jiang, Anja R. Ollodart and Maitreya J. Dunham, date: 06/05/2022, view: 697, Q&A: 0
    [Abstract]

    Understanding the generation of mutations is fundamental to understanding evolution and genetic disease; however, the rarity of such events makes experimentally identifying them difficult. Mutation accumulation (MA) methods have been widely used. MA lines require serial bottlenecks to fix de novo mutations, followed by whole-genome sequencing.

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    Efficient Superovulation and Egg Collection from Mice
    Authors:  Miyuki Shindo, Kenji Miyado, Woojin Kang, Maki Fukami and Mami Miyado, date: 06/05/2022, view: 736, Q&A: 0
    [Abstract]

    Superovulation is a method used to reduce the number of mice used per experiment by increasing the egg number. Conventionally, superovulation for obtaining mouse eggs involves the use of equine chorionic gonadotropin (eCG) for stimulation and human CG for induction. Female mice of the C57BL/6 inbred strain spontaneously ovulate approximately 10

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