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Press Needle: A Traditional Chinese Medicine Therapy for Myopia Patients with Dry Eye
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Cited by 2 •
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Here, we present a protocol to standardize the operation of acupuncture in the treatment of myopia with dry eye.
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Detection of Disease-associated α-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated α-synuclein
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An ELISA offering a novel quantitative approach is described. It specifically detects disease-associated α-synuclein (αSD) in a transgenic mouse model (M83) of synucleinopathy using several antibodies against either the Ser129 phosphorylated αS form or the C-terminal part of the protein.
08:43
Isolation and Characterization of Patient-derived Pancreatic Ductal Adenocarcinoma Organoid Models
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Patient-derived organoid cultures of pancreatic ductal adenocarcinoma are a rapidly established 3-dimensional model that represent epithelial tumor cell compartments with high fidelity, enabling translational research into this lethal malignancy. Here, we provide detailed methods to establish and propagate organoids as well as to perform relevant biological assays using these models.
09:04
Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
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This protocol provides a method to facilitate the generation of defined heterozygous or homozygous nucleotide changes using CRISPR-CAS9 in human pluripotent stem cells.
07:05
Operant Protocols for Assessing the Cost-benefit Analysis During Reinforced Decision Making by Rodents
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Cited by 3 •
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A cost-benefit analysis is a weighing-scale approach that the brain performs during the course of decision making. Here, we propose a protocol to train rats on an operant-based decision-making paradigm where rats choose higher rewards at the expense of waiting for 15 s to receive them.
FAQs
The concentrated suspension passes through a narrow valve or orifice under high pressure, then experiences a rapid pressure drop as it exits. This transition generates intense shear stress that disrupts cell envelopes and releases intracellular material. The mechanical action is central to obtaining lysates without relying on chemical detergents.
Mechanical disruption can produce heat, which may damage temperature-sensitive cellular components or reduce biochemical activity. Temperature control helps limit this heat-related damage while the suspension undergoes pressurization and rapid release. Maintaining suitable conditions is therefore important when the lysate will be used for enzyme assays, protein analysis, or other activity-dependent workflows.
French Press lysis uses mechanical forces rather than chemical detergents to break open cells. Avoiding detergents can help preserve biochemical activity and simplify downstream purification, because the lysate is not complicated by those chemical additives. This distinction is especially relevant when researchers plan to purify proteins or examine enzyme function after disruption.
Several conditions affect the resulting lysate: the cell suspension must be concentrated, the material must pass through a narrow valve or orifice, and pressure release must generate sufficient shear stress for envelope disruption. Temperature control also matters because excessive heat can damage released components. Together, these conditions influence the usefulness of the preparation for later analysis.
A typical workflow begins with a concentrated suspension of biological cells, followed by passage through the disruptor’s narrow valve or orifice under pressure. The released material is then commonly subjected to centrifugation to separate components before chromatography, enzyme assays, or molecular biology procedures. Temperature control is maintained during disruption to limit heat-related damage.
Researchers may choose this approach when they need lysates from bacteria, yeast, or other microorganisms and want to recover intracellular proteins, nucleic acids, or organelles. It is useful before centrifugation, chromatography, enzyme assays, and molecular biology workflows. In biology, the method connects physical cell disruption with characterization or purification of intracellular material.