scholarly journals Detection of silver nanoparticles in cells by flow cytometry using light scatter and far-red fluorescence

2013 ◽  
pp. n/a-n/a ◽  
Author(s):  
R. M. Zucker ◽  
K. M. Daniel ◽  
E. J. Massaro ◽  
S. J. Karafas ◽  
L. L. Degn ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Silver Nanoparticles ◽  
Light Scatter ◽  
Red Fluorescence ◽  
In Cells

Detection of intracellular silver nanoparticles using flow cytometry

2018 ◽  
pp. 223-226
Author(s):  
С.И. Каба ◽  
А.А. Соколовская
Keyword(s):  
Flow Cytometry ◽  
Silver Nanoparticles ◽  
Endothelial Cells ◽  
Silver Nanoparticle ◽  
Cell Uptake ◽  
Control Samples

Продемонстрировано обнаружение наночастиц серебра во внутриклеточном пространстве с помощью проточной цитофлуориметрии. В эндотелиальных клетках линии EA.hy926, инкубированных в растворе, содержащем 2 мкг/мл наносеребра, измеряли боковое светорассеяние. По сравнению с контрольными образцами этот параметр возрастал, в то время как прочие значимые характеристики не изменялись. Это подтверждает чувствительность метода к изменившемуся состоянию клеток и указывает на поглощение наночастиц серебра клетками при концентрации ниже токсической. The study demonstrated a possibility for detection of intracellular silver nanoparticles using flow cytometry. The parameter used in this work, side scattering, was measured in EA.hy926 endothelial cells incubated in a 2 mg/ml silver nanoparticle solution. This parameter was increased compared to control samples. Therefore, this technique was sensitive to changes in the cell status and suggested the cell uptake of the particles under the subtoxic conditions.

Annona muricata silver nanoparticles exhibit strong anticancer activities against cervical and prostate adenocarcinomas through regulation of CASP9 and the CXCL1/CXCR2 genes axis

Tumor Biology ◽  
2021 ◽  
Vol 43 (1) ◽  
pp. 37-55
Author(s):  
Yahaya Gavamukulya ◽  
Esther N. Maina ◽  
Hany A. El-Shemy ◽  
Amos M. Meroka ◽  
Geoffrey K. Kangogo ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Anticancer Drugs ◽  
Hela Cells ◽  
Selectivity Index ◽  
Annona Muricata ◽  
Anticancer Activities ◽  
Apoptosis Pathway ◽  
Migration Assay ◽  
Intrinsic Apoptosis Pathway ◽  
In Cells

BACKGROUND: Green synthesized nanoparticles have been earmarked for use in nanomedicine including for the development of better anticancer drugs. OBJECTIVE: The aim of this study was to undertake biochemical evaluation of anticancer activities of green synthesized silver nanoparticles (AgNPs) from ethanolic extracts of fruits (AgNPs-F) and leaves (AgNPs-L) of Annona muricata. METHODS: Previously synthesized silver nanoparticles were used for the study. The effects of the AgNPs and 5-Fluorouracil were studied on PC3, HeLa and PNT1A cells. The resazurin, migration and colonogenic assays as well as qRT-PCR were employed. RESULTS: The AgNPs-F displayed significant antiproliferative effects against HeLa cells with an IC50 of 38.58μg/ml and PC3 cells with an IC50 of 48.17μg/ml but selectively spared normal PNT1A cells (selectivity index of 7.8), in comparison with first line drug 5FU and AgNPs-L whose selectivity index were 3.56 and 2.26 respectively. The migration assay revealed potential inhibition of the metastatic activity of the cells by the AgNPs-F while the colonogenic assay indicated the permanent effect of the AgNPs-F on the cancer cells yet being reversible on the normal cells in contrast with 5FU and AgNPs-L. CASP9 was significantly over expressed in all HeLa cells treated with the AgNPs-F (1.53-fold), AgNPs-L (1.52-fold) and 5FU (4.30-fold). CXCL1 was under expressed in HeLa cells treated with AgNPs-F (0.69-fold) and AgNPs-L (0.58-fold) and over expressed in cells treated with 5FU (4.95-fold), but the difference was not statistically significant. CXCR2 was significantly over expressed in HeLa cells treated with 5FU (8.66-fold) and AgNPs-F (1.12-fold) but under expressed in cells treated with AgNPs-L (0.76-fold). CONCLUSIONS: Here we show that biosynthesized AgNPs especially AgNPs-F can be used in the development of novel and better anticancer drugs. The mechanism of action of the AgNPs involves activation of the intrinsic apoptosis pathway through upregulation of CASP9 and concerted down regulation of the CXCL1/ CXCR2 gene axis.

scholarly journals Tracking protein aggregation and mislocalization in cells with flow cytometry

2012 ◽  
Vol 9 (5) ◽  
pp. 467-470 ◽  
Author(s):  
Yasmin M Ramdzan ◽  
Saskia Polling ◽  
Cheryl P Z Chia ◽  
Ivan H W Ng ◽  
Angelique R Ormsby ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Protein Aggregation ◽  
In Cells

scholarly journals A Multiplexed Assay That Monitors Effects of Multiple Compound Treatment Times Reveals Candidate Immune-Enhancing Compounds

2018 ◽  
Vol 23 (7) ◽  
pp. 646-655 ◽  
Author(s):  
Ziyan Zhao ◽  
Liza Henowitz ◽  
Adam Zweifach
Keyword(s):  
Flow Cytometry ◽  
Cytotoxic T Lymphocytes ◽  
High Throughput ◽  
Treatment Time ◽  
Single Sample ◽  
Light Scatter ◽  
Flow Cytometry Assay ◽  
Multiple Treatment ◽  
Lytic Granule ◽  
Activation Status

We previously developed a flow cytometry assay that monitored lytic granule exocytosis in cytotoxic T lymphocytes stimulated by contacting beads coated with activating anti-CD3 antibodies. That assay was multiplexed in that responses of cells that did or did not receive the activating stimulus were distinguished via changes in light scatter accompanying binding of cells to beads, allowing us to discriminate compounds that activate responses on their own from compounds that enhance responses in cells that received the activating stimulus, all within a single sample. Here we add a second dimension of multiplexing by developing means to assess in a single sample the effects of treating cells with test compounds for different times. Bar-coding cells before adding them to test wells lets us determine compound treatment time while also monitoring activation status and response amplitude at the point of interrogation. This multiplexed assay is suitable for screening 96-well plates. We used it to screen compounds from the National Cancer Institute, identifying several compounds that enhance anti-LAMP1 responses. Multiple-treatment-time (MTT) screening enabled by bar-coding and read via high-throughput flow cytometry may be a generally useful method for facilitating the discovery of compounds of interest.

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