Detection of Apoptotic Cells Using Propidium Iodide Staining

2014 ◽  
Vol 2014 (11) ◽  
pp. pdb.prot082545 ◽  
Author(s):  
Andrea Newbold ◽  
Ben P. Martin ◽  
Carleen Cullinane ◽  
Michael Bots
Keyword(s):  
Propidium Iodide ◽  
Apoptotic Cells ◽  
Propidium Iodide Staining

scholarly journals Corrigendum: Detection of Apoptotic Cells Using Propidium Iodide Staining

2018 ◽  
Vol 2018 (6) ◽  
pp. Corr104943
Author(s):  
Andrea Newbold ◽  
Ben P. Martin ◽  
Carleen Cullinane ◽  
Michael Bots
Keyword(s):  
Propidium Iodide ◽  
Apoptotic Cells ◽  
Propidium Iodide Staining

scholarly journals Visual Recognition and Efficient Isolation of Apoptotic Cells with Fluorescent-Magnetic-Biotargeting Multifunctional Nanospheres

2007 ◽  
Vol 53 (12) ◽  
pp. 2177-2185 ◽  
Author(s):  
Er-Qun Song ◽  
Guo-Ping Wang ◽  
Hai-Yan Xie ◽  
Zhi-Ling Zhang ◽  
Jun Hu ◽  
...  
Keyword(s):  
Ultraviolet Light ◽  
Clinical Diagnosis ◽  
Propidium Iodide ◽  
Biomedical Applications ◽  
Visual Recognition ◽  
Nuclear Dna ◽  
Annexin V ◽  
Living Cells ◽  
Apoptotic Cells ◽  
Propidium Iodide Staining

Abstract Background: Fluorescent-magnetic-biotargeting multifunctional nanospheres are likely to find important applications in bioanalysis, biomedicine, and clinical diagnosis. We have been developing such multifunctional nanospheres for biomedical applications. Methods: We covalently coupled avidin onto the surfaces of fluorescent-magnetic bifunctional nanospheres to construct fluorescent-magnetic-biotargeting trifunctional nanospheres and analyzed the functionality and specificity of these trifunctional nanospheres for their ability to recognize and isolate apoptotic cells labeled with biotinylated annexin V, which recognizes phosphatidylserine exposed on the surfaces of apoptotic cells. Results: The multifunctional nanospheres can be used in combination with propidium iodide staining of nuclear DNA to identify cells at different phases of the apoptotic process. Furthermore, we demonstrate that apoptotic cells induced by exposure to ultraviolet light can be isolated simply with a magnet from living cells at an efficiency of at least 80%; these cells can then be easily visualized with a fluorescence microscope. Conclusions: Our results show that fluorescent-magnetic-biotargeting trifunctional nanospheres can be a powerful tool for rapidly recognizing, magnetically enriching and sorting, and simultaneously identifying different kinds of cells.

High‐Throughput Cytotoxicity Screening by Propidium Iodide Staining

2007 ◽  
Vol 41 (1) ◽  
Author(s):  
Bruce S. Edwards ◽  
Irena Ivnitski‐Steele ◽  
Susan M. Young ◽  
Virginia M. Salas ◽  
Larry A. Sklar
Keyword(s):  
High Throughput ◽  
Propidium Iodide ◽  
Propidium Iodide Staining

Annexin V staining during reperfusion detects cardiomyocytes with unique properties

2001 ◽  
Vol 281 (5) ◽  
pp. H1931-H1937 ◽  
Author(s):  
Prakash Narayan ◽  
Robert M. Mentzer ◽  
Robert D. Lasley
Keyword(s):  
Propidium Iodide ◽  
Ventricular Myocytes ◽  
Contractile Function ◽  
Ischemia Reperfusion ◽  
Annexin V ◽  
Cell Width ◽  
Simulated Ischemia ◽  
Propidium Iodide Staining ◽  
Membrane Blebs ◽  
Apoptosis And Necrosis

With the use of markers of sarcolemmal membrane permeability, cardiomyocyte models of ischemic injury have primarily addressed necrotic death during ischemia. In the present study, we used annexin V-propidium iodide staining to examine apoptosis and necrosis after simulated ischemia and simulated reperfusion in rat ventricular myocytes. Annexin V binds phosphatidylserine, a phosphoaminolipid thought to be externalized during apoptosis or programmed cell death. Propidium iodide is a marker of cell necrosis. Under baseline conditions, <1% of cardiomyocytes stained positive for annexin V. After 20 or 60 min of simulated ischemia, there was no increase in annexin V staining, although 60-min simulated ischemia resulted in significant propidium iodide staining. Twenty minutes of simulated ischemia, followed by 20 or 60 min of simulated reperfusion, resulted in 8–10% of myocytes staining positive for annexin V. Annexin V-positive cells retained both rod-shaped morphology and contractile function but exhibited the decreased cell width indicative of cell shrinkage. Baseline mitochondrial free Ca2+(111 ± 14 nM) was elevated in reperfused annexin V-negative cells (214 ± 22 nM), and further elevated in annexin V-positive myocytes (382 ± 9 nM). After 60 min of simulated reperfusion, caspase-3-like activity was observed in ∼3% of myocytes, which had a rounded appearance and membrane blebs. These results suggest that the use of annexin V after simulated ischemia-reperfusion uncovers a population of cardiomyocytes whose characteristics appear to be consistent with cells undergoing apoptosis.

A comparison of fluorescein diacetate and propidium iodide staining andin vitroexcystation for determiningGiardia intestinaliscyst viability

Parasitology ◽  
1989 ◽  
Vol 99 (3) ◽  
pp. 329-331 ◽  
Author(s):  
A. L. Smith ◽  
H. V. Smith
Keyword(s):  
Lower Limit ◽  
Propidium Iodide ◽  
Giardia Intestinalis ◽  
Fluorescein Diacetate ◽  
Fluorescent Indicators ◽  
Vital Dyes ◽  
Propidium Iodide Staining ◽  
Cyst Viability

SummaryThe viability of 4 human isolates ofGiardia intestinaliscysts using either the fluorogenic vital dyes fluorescein diacetate (FDA) and propidium iodide (PI) orin vitroexcystation was assessed. Whereas viable cysts, as defined byin vitroexcystation were present in each of the 4 isolates, cysts from only 3 of the 4 isolates took up the vital dyes. FDA consistently over-estimated cyst viability whilst PI under-estimated non-viable cysts when compared within vitroexcystation. Followingin vitroexcystation, both FDA and PI stained a proportion of unexcysted cysts indicating that FDA stained cysts which were incapable of excystation, whereas PI did not stain all cysts which were incapable of excystation. One human cyst isolate, which underwentin vitroexcystation, could not be stained with either FDA or PI. In the absence of currently more specific fluorescent indicators of viability, PI alone could be used to determine the lower limit of nonviability in positive water-related samples, where small numbers of cysts are to be expected.

scholarly journals Microampere electric currents caused bacterial membrane damage and two-way leakage in short time

2020 ◽  
Author(s):  
V R Krishnamurthi ◽  
A Rogers ◽  
J Peifer ◽  
I Niyonshuti ◽  
J Chen ◽  
...  
Keyword(s):  
Small Molecules ◽  
Single Molecule ◽  
Propidium Iodide ◽  
Membrane Damage ◽  
Bacterial Membrane ◽  
Electric Currents ◽  
Localization Microscopy ◽  
Propidium Iodide Staining ◽  
Short Time ◽  
Single Molecule Localization Microscopy

AbstractPhysical agents such as low electric voltages and currents have recently gained attention for antimicrobial treatment due to their bactericidal capability. Although microampere electric currents were shown to suppress the growth of bacteria, it remains unclear to what extent the microampere currents damage bacterial membrane. Here, we investigated the membrane damage and two-way leakage caused by microampere electric currents (≤ 100 μA) in a short time (30 min). Based on MitoTracker staining, propidium iodide staining, filtration assays, and quantitative single-molecule localization microscopy, we found that microampere electric currents caused significant membrane damages and allowed two-way leakages of ions, small molecules and proteins. This study paves the way to new development and antibiotic applications of ultra-low electric voltages and currents.Statement of SignificancePrevious studies showed that treating bacteria with milliampere electric currents for 72 hours led to significant damages of the bacterial membrane. However, it remains unclear to what extent membrane damages and two-way (i.e. inward and outward) leakages are caused by lower electric currents in a shorter time. In this work, we set out to answer this question. We carried out several assays on the bacteria treated by microampere electric currents of ≤ 100 μA for 30 min, including MitoTracker staining, propidium iodide staining, filtration assays, and quantitative single-molecule localization microscopy. We found and quantified that the membrane damages were caused by microampere electric currents in half an hour and allowed two-way leakages of ions, small molecules, and proteins.

scholarly journals The Role of Laundry Detergents in the Elements of Skin Pathogen

2021 ◽  
Author(s):  
Erdong Wei ◽  
Markus Reinholz ◽  
Lars E. French ◽  
Benjamin M. Clanner-Engelshofen
Keyword(s):  
Propidium Iodide ◽  
Laundry Detergent ◽  
Antimicrobial Effect ◽  
Antimicrobial Efficacy ◽  
Liquid Form ◽  
Laundry Detergents ◽  
Propidium Iodide Staining ◽  
Bleach Activator ◽  
Antibacterial And Antifungal ◽  
Skin Pathogen

Abstract Background: The Corona pandemic fuelled up skin pathogen challenges in young and adults, the antimicrobial efficacy of laundry detergents could be considered particularly. However, no available data focusing on the form of laundry detergent, additives and conditions affect the antimicrobial efficacy. This study simulated washing procedures to investigate the antibacterial and antifungal activity of laundry detergents.Methods and Results: Mimic laundry procedures were performed to treat Gram-positive bacteria, Gram-negative bacteria and fungus, colony counting and propidium iodide staining were used to assess the antimicrobial activity. Powder detergent A, NaBO3*4H2O with the tetraacetylethylenediamine, 2Na2co3.3H2O with tetraacetylethylenediamine could achieve a > 5-log10 reduction of three microbial colony generation. Anionic surfactant sodium dodecylbenzene sulphonate (SDBS) group had the strongest fluorescence intensity in three microbial propidium iodide staining.Conclusions: Powder form laundry detergents are superior to liquid form, peroxide-based bleaches and bleach activator in solid form, the solid surfactants with matched PH and alkyl chain length showed a considerable antimicrobial effect.

scholarly journals Effect and Mechanism of Survivin on Hypoxia-Induced Multidrug Resistance of Human Laryngeal Carcinoma Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-6
Author(s):  
Dan Xu ◽  
Da Wei Li ◽  
Jin Xie ◽  
Xin Wei Chen
Keyword(s):  
Multidrug Resistance ◽  
Rna Interference ◽  
Cancer Cells ◽  
Propidium Iodide ◽  
Laryngeal Carcinoma ◽  
Survivin Expression ◽  
Annexin V ◽  
Carcinoma Cells ◽  
Propidium Iodide Staining ◽  
Human Laryngeal Carcinoma

This study aimed at clarifying the mechanism and role of survivin in hypoxia-induced multidrug resistance (MDR) of laryngeal carcinoma cells. Human laryngeal cancer cells were incubated under hypoxia or normoxia. The expression of survivin was silenced by performing RNA interference. Additionally, by Western blot and real-time quantitative RT-PCR, survivin expression was detected. The sensitivity of human laryngeal carcinoma cells to multiple drugs was measured by CCK-8 assay. Meanwhile, the apoptosis of cells induced by cisplatin or paclitaxel was assessed by Annexin-V/propidium iodide staining analysis. Under hypoxic conditions, the upregulation of survivin was abolished by RNA interference. Then, CCK-8 analysis demonstrated that the sensitivity to multiple agents of laryngeal carcinoma cells could be increased by inhibiting survivin expression (P<0.05). Moreover, Annexin-V/propidium iodide staining analysis revealed that decreased expression of survivin could evidently increase the apoptosis rate of laryngeal carcinoma cells that were induced by cisplatin or paclitaxel evidently (P<0.05). Our data suggests that hypoxia-elicited survivin may exert a pivotal role in regulating hypoxia-induced MDR of laryngeal cancer cells by preventing the apoptosis of cells induced by chemotherapeutic drug. Thus, blocking survivin expression in human laryngeal carcinoma cells may provide an avenue for gene therapy.

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