Plasma Membrane (PM) Repair
Cells are equipped with multiple PM repair mechanisms to rapidly reseal the PM, thereby restoring its barrier function and maintaining cell survival. Those covering ion imbalance lead to cytoskeletal remodeling, reduction of PM tension, repair factor recruitment and vesicle transport to collectively promote PM repair. Although several different repair pathways have been identified above, our understanding of how these mechanisms cooperate to facilitate resealing remains unclear. Furthermore, excessive damage to the PM and its repair defects have been associated with pathological conditions such as infection, myotonic dystrophy, neurodegenerative diseases, etc. Therefore, in order to PM the various steps and molecular factors involved in the repair process, it is necessary to use sensitive and rapid assays in order to accurately measure the efficiency of repair.
Fig. 1. Plasma membrane damage and repair in the liver, pancreas, nervous system, kidney, and muscle. (Ammendolia D A, et al., 2021)
Services
Measuring the extent of PM repair is an important indicator of the lysosomal cytosolic action. CD BioSciences is committed to developing high-throughput, microtiter plate-based methods to monitor the efficiency of PM resealing. Our measurement service for the extent of PM repair is widely used to identify the cellular components that control plasma membrane resealing and the drugs that improve resealing.
Here, we provide the following methods that rely on the use of membrane impermeable dyes for quantification of PM integrity. Based on the advantages and disadvantages of both methods, we will develop the best solution for your project.
Propidium iodide (PI) Influx
PI produces quantifiable fluorescence upon binding to nucleic acids in the cell. We use PI to label the PM of cells and measure the uptake of the fluorescent dye by damaged cells by quantitative fluorescence microscopy and flow cytometry to quantify the efficiency of PM.
- PI is not fixable and microscopic imaging is performed as soon as possible after the assay is completed.
- Flow cytometry allows rapid measurement of large cell populations and is well suited for suspension cells.
- Quantitative fluorescence microscopy is suitable for apposed mammalian cells and is not suitable for high throughput analysis.
- A Fast, sensitive measurement tool.
- High-throughput analysis.
- Uses a temperature-controlled multi-mode zymography.
- Can be used to compare resealing efficiency of primary cells and cell lines from different sources.
Membrane Selective Lipophilic FM Dyes
FM4-64 and FM1-43 can be reversibly partitioned to the outer leaflet of the cell membrane without permeabilization. We use FM4-64 and FM1-43 to label intracellular membranes to determine the kinetics of cell damage and reseal in real time by combining microscopic real-time imaging with image analysis.
- Real-time imaging of membrane repair is achieved.
- Can help identify differences between important players and regulators of membrane repair.
- FM1-43 dye can also be added at the time of imaging.
- Unable to determine the kinetics of PM repair.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
References
- Ammendolia D A, Bement W M, Brumell J H. (2021) Plasma membrane integrity: implications for health and disease[J]. BMC biology. 19(1): 1-29.
- Blazek AD, Paleo BJ, Weisleder N. (2015) Plasma Membrane Repair: A Central Process for Maintaining Cellular Homeostasis. Physiology (Bethesda). 30(6):438-48.