Bacterial Lysis Assay
Bacterial Lysis Assay - Related Videos
03:12
A Procedure for Bacterial Harvesting and Mechanical Lysis
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2025
Source: Mercier, N., et al. MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria. J. Vis. Exp. (2021)This video demonstrates a step-by-step procedure for harvesting bacterial cells and mechanically lysing them to release intracellular contents.
04:02
An Assay to Measure the Accumulation of a Fluorescent Antibiotic Probe in Bacterial Cells
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2025
This video demonstrates an assay to measure the accumulation of a fluorescent antibiotic probe in bacterial cells. Exposure to an efflux pump inhibitor disrupts the transmembrane proton gradient in bacterial cells, upon which a fluorophore-conjugated antibiotic is added. Inhibition of efflux pumps due to the disrupted proton gradient causes accumulation of the antibiotic inside the bacteria. Bacterial cells are lysed, and the fluorescence intensity is measured to confirm antibiotic accumulation.
05:28
A Bacterial Infection Assay for Studying Anaerobic Bacteria-Host Cell Interactions
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2025
This video demonstrates a technique for assessing the ability of Porphyromonas gingivalis, an anaerobic pathogenic bacterium, to adhere to and invade human umbilical vein endothelial cells (HUVECs). After infecting the host cells with the bacteria, the host cells are lysed to release both adhered and internalized bacteria. When the lysate is plated in an appropriate medium, the presence of colonies indicates successful bacteria-host cell interactions.
02:46
Swarming Assay for Visualizing Bacterial Surface Motility
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2025
Source: Morales-Soto, N. et. al., Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays. J. Vis. Exp. (2015)This video demonstrates a swarming assay using bacteria that express green fluorescent protein to observe surface motility on soft agar. Time-lapse imaging captures coordinated expansion and tendril formation, highlighting bacterial surface motility.
07:35
Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays
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Cited by 68 •
2015
Swarming motility is influenced by physical and environmental factors. We describe a two-phase protocol and guidelines to circumvent the challenges commonly associated with swarm assay preparation and data collection. A macroscopic imaging technique is employed to obtain detailed information on swarm behavior that is not provided by current analysis techniques.
FAQs
Cell-envelope disruption compromises the boundary that retains bacterial contents. As structural damage progresses, intracellular components escape into the surrounding medium, altering the optical properties of the culture. This connection allows the assay to translate a physical event, loss of cellular integrity, into a measurable change in turbidity or absorbance.
Turbidity and absorbance provide indirect measures of changes occurring throughout a bacterial culture. When treatment-associated lysis changes the amount or distribution of cellular material, the culture can show a measurable optical shift, often a decrease. These readouts therefore help researchers compare the extent of disruption produced under different treatment conditions.
Different treatments can be examined according to how strongly they disrupt bacterial integrity and how the culture responds optically. Antibiotics, antimicrobial compounds, bacteriophages, and lytic enzymes may therefore be compared within the same experimental framework. Differences in the measured response help characterize antibacterial action without treating all reductions in bacterial survival as mechanistically identical.
A typical workflow exposes a bacterial culture to a treatment or condition of interest and then monitors the culture for evidence of cell disruption. The outcome is assessed through turbidity or absorbance changes associated with loss of integrity and release of intracellular material. Comparing the measured responses provides an estimate of relative treatment effectiveness.
The assay supports evaluation of several antibacterial or lytic agents, including antibiotics, antimicrobial compounds, bacteriophages, and lytic enzymes. It can also examine other conditions that affect bacterial survival. This breadth makes the method useful when the goal is to compare treatments, identify strong disruptive effects, or investigate how a condition influences bacterial integrity.
In biology, results can support studies of antibacterial mechanisms, treatment effectiveness, and host-microbe interactions. The measurements are also relevant to microbiology, biotechnology, and infectious disease research, where researchers need evidence of altered bacterial integrity or survival. By linking treatment exposure with an observable culture response, the assay contributes to broader analyses of bacterial behavior.