Bullseye Analysis: A Fluorescence Microscopy Technique to Detect Local Changes in Intracellular Reactive Oxygen Species (ROS) Production
نویسندگان
چکیده
Abstract Reactive oxygen species (ROS) are naturally produced compounds that play important roles in cell signaling, gene regulation, and biological defense, including involvement the oxidative burst is central to anti-microbial actions of macrophages. However, these highly reactive, short-lived radical also stimulate cells undergo programmed death at high concentrations, as well causing detrimental effects such oxidation macromolecules more moderate levels. Imaging ROS challenging, with many researchers working on challenge over past 10–15 years without producing a definitive method. We report new fluorescence microscopy-based technique, Bullseye Analysis. This methodology based concepts provided by FRAP (Fluorescence Recovery after Photobleaching) technique refined evidence spatiotemporal production ROS, subsequent consequences, subcellular scale. To exemplify we have used ROS-reporter dye, CellROX, ROS-inducing photosensitizer, LightOx58, potent source compared UV irradiation alone. Further validation was carried out using differing co-stains, notably Mitotracker JC-1.
منابع مشابه
Reactive Oxygen Species (Ros-Induced) Ros Release
We sought to understand the relationship between reactive oxygen species (ROS) and the mitochondrial permeability transition (MPT) in cardiac myocytes based on the observation of increased ROS production at sites of spontaneously deenergized mitochondria. We devised a new model enabling incremental ROS accumulation in individual mitochondria in isolated cardiac myocytes via photoactivation of t...
متن کاملProduction and detection of reactive oxygen species (ROS) in cancers.
Reactive oxygen species include a number of molecules that damage DNA and RNA and oxidize proteins and lipids (lipid peroxydation). These reactive molecules contain an oxygen and include H(2;)O(2;) (hydrogen peroxide), NO (nitric oxide), O(2;)(-) (oxide anion), peroxynitrite (ONOO(-)), hydrochlorous acid (HOCl), and hydroxyl radical (OH(-)). Oxidative species are produced not only under patholo...
متن کاملP-122: Sperm Intracellular Reactive Oxygen Species Incidence in Ram Fed A Diet with Fish Oil
Background: Basal level of reactive oxygen species (ROS) is essential for sperm acrosomal reaction, hyperactivation, motility, capacitation and fertilization. Ram spermatozoa contain large quantities of polyunsaturated fatty acids in their plasma membranes so are extremely susceptible to ROS induced damage. Materials and Methods: Eight rams (36 ± 5 month of age and with a weight of 64 ± 7 kg) w...
متن کاملMitochondrial reactive oxygen species (ROS) and ROS-induced ROS release.
Byproducts of normal mitochondrial metabolism and homeostasis include the buildup of potentially damaging levels of reactive oxygen species (ROS), Ca(2+), etc., which must be normalized. Evidence suggests that brief mitochondrial permeability transition pore (mPTP) openings play an important physiological role maintaining healthy mitochondria homeostasis. Adaptive and maladaptive responses to r...
متن کاملReactive Oxygen Species (Ros) Generation in Sepsis
Sepsis and septic shock remain as leading cause of death in adult intensive care units. It is widely accepted that sepsis and septic shock are caused predominantly by gram-negative bacteria and their endotoxins. Endotoxin or Lipopolysaccharide (LPS) have important roles as host responses and trigger the inflammatory processes, caused by gram-negative bacterial infection. Production of oxygen ra...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Microscopy and Microanalysis
سال: 2022
ISSN: ['1435-8115', '1431-9276']
DOI: https://doi.org/10.1093/micmic/ozac040