scholarly journals Evaluation of PicoGreen variants for use in microscopy and flow cytometry

2019 ◽  
Author(s):  
Muznah Khatoo ◽  
Jongtae Yang ◽  
Kyle R. Gee ◽  
Stephan Gasser
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Fluorescent Probe ◽  
Fluorescent Microscopy ◽  
Dna Quantitation ◽  
Unique Probe ◽  
Limited Application ◽  
Low Levels

AbstractPicoGreen is a fluorescent probe that binds dsDNA and forms a highly luminescent complex when compared to the free dye in solution. This unique probe is widely used in DNA quantitation assays but has limited application in flow cytometry and microscopy. Here we have investigated various PicoGreen variants for the ability to stain low amounts of cytosolic DNA present in many tumor cells. Analysis of stained cells by flow cytometry and fluorescent microscopy showed that certain variants improved the ability to stain low levels of cytosolic DNA when compared to the commercially available PicoGreen molecule.

scholarly journals Quantifying circulating Th17 cells by qPCR: potential as diagnostic biomarker for rheumatoid arthritis

Rheumatology ◽  
2019 ◽  
Vol 58 (11) ◽  
pp. 2015-2024 ◽  
Author(s):  
Agata N Burska ◽  
Aye Thu ◽  
Rekha Parmar ◽  
Izabella Bzoma ◽  
Bjoern Samans ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Th17 Cells ◽  
Regression Models ◽  
Qpcr Assay ◽  
T Cell Subsets ◽  
Correct Prediction ◽  
Diagnostic Biomarker ◽  
Negative Group ◽  
Disease Site ◽  
Low Levels

Abstract Objective The diagnosis of RA patients remains a challenge, especially in ACPA-negative disease. Novel T-cell subsets, particularly Th17 may be useful, although data on Th17 frequency using flow cytometry in RA are conflicting. We investigated whether a novel epigenetic qPCR assay for the quantification of Th17 could differentiate patients with RA from those with symptoms evolving towards an alternative diagnosis. Methods We used a qPCR assay measuring the extent of the methylation at a key position in the IL-17 and CD4 genes. Assays were performed on whole blood from 49 healthy controls (HC) and 165 early arthritis clinic patients. Flow cytometry was further used to detect the expression of CXCR4 on Th17 cells. Results In 75 inflammatory arthritis patients who progressed to RA, the qPCR assays showed significantly fewer Th17 cells compared with 90 patients who did not (P<0.0001). Regression models demonstrated a high predictive value for RA development (75.8% correct prediction), and particularly for the ACPA-negative group (n = 125) where Th17 and swollen joint count (SJC) were the only predictors (73% correct prediction). The chemokine receptor CXCR4 had significantly higher expression on Th17 from early RA patients (n = 11) compared with HC (n = 15). Conclusion The results of the epigenetic qPCR assay showed that low levels of Th17 cells were predictive of developing RA, particularly in the ACPA-negative patients. This could have value for insights into pathogenesis and management. The results suggest the recruitment of Th17 to the inflammatory disease site, consistent with high CXCR4 expression.

scholarly journals Interleukin 2 gene transfer into tumor cells abrogates tumorigenicity and induces protective immunity.

1990 ◽  
Vol 172 (4) ◽  
pp. 1217-1224 ◽  
Author(s):  
B Gansbacher ◽  
K Zier ◽  
B Daniels ◽  
K Cronin ◽  
R Bannerji ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Tumor Cells ◽  
Interleukin 2 ◽  
Retroviral Vectors ◽  
Cytolytic Activity ◽  
Tumor Formation ◽  
Tumor Bearing ◽  
Low Levels ◽  
Parental Tumor ◽  
Observation Period

To study the effects of localized secretion of cytokines on tumor progression, the gene for human interleukin 2 (IL-2) was introduced via retroviral vectors into CMS-5 cells, a weakly immunogenic mouse fibrosarcoma cell line of BALB/c origin. Secretion of low levels of IL-2 from the tumor cells abrogated their tumorigenicity and induced a long-lasting protective immune response against a challenge with a tumorigenic dose of parental CMS-5 cells. Co-injection of IL-2-producing CMS-5 cells with unmodified tumor cells inhibited tumor formation even when highly tumorigenic doses of CMS-5 cells were used. Cytolytic activity in mice injected with parental CMS-5 cells was transient and was greatly diminished 3 wk after injection, as commonly observed in tumor-bearing animals. However, in mice injected with IL-2-producing cells, tumor-specific cytolytic activity persisted at high levels for the duration of the observation period (at least 75 d). High levels of tumor-specific cytolytic activity could also be detected in parental CMS-5 tumor-bearing animals 18 d after inoculation with tumor cells, if IL-2-producing CMS-5 cells but not unmodified parental tumor cells were used as targets. These studies highlight the potential advantages of localized secretion of cytokines mediated via gene transfer to induce potent anti-tumor immune responses.

A cyanine-derived “turn-on” fluorescent probe for imaging nitroreductase in hypoxic tumor cells

2015 ◽  
Vol 7 (24) ◽  
pp. 10125-10128 ◽  
Author(s):  
Cong Xue ◽  
Yingjie Lei ◽  
Sichun Zhang ◽  
Yaowu Sha
Keyword(s):  
Tumor Cells ◽  
Fluorescent Probe ◽  
Phenol Group ◽  
Turn On ◽  
Hypoxic Tumor

A new “turn-on” fluorescent probe, composed of a protected phenol group with ap-nitrobenzyl moiety that functions as a latent donor and conjugated with two benzo[f]indolinium acceptors, was developed and applied for imaging nitroreductase (NTR) in hypoxic tumor cells.

scholarly journals Postoperative metastasis prediction based on portal vein circulating tumor cells detected by flow cytometry in periampullary or pancreatic cancer

2019 ◽  
Vol Volume 11 ◽  
pp. 7405-7425 ◽  
Author(s):  
Lianyuan Tao ◽  
Li Su ◽  
Chunhui Yuan ◽  
Zhaolai Ma ◽  
Lingfu Zhang ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Flow Cytometry ◽  
Portal Vein ◽  
Tumor Cells ◽  
Circulating Tumor Cells

scholarly journals In vivo quantitation of rare circulating tumor cells by multiphoton intravital flow cytometry

2007 ◽  
Vol 104 (28) ◽  
pp. 11760-11765 ◽  
Author(s):  
W. He ◽  
H. Wang ◽  
L. C. Hartmann ◽  
J.-X. Cheng ◽  
P. S. Low
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Circulating Tumor Cells

scholarly journals Determination of Circulating Tumor Cells by Flow Cytometry in the Bladder Cancer Patients

2018 ◽  
Vol 2 (1) ◽  
pp. 25-30 ◽  
Author(s):  
Elif Ercan ◽  
◽  
Ender Sımsek ◽  
Ozen Ozensoy Guler ◽  
Abdullah Erdem Canda ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bladder Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells

A melatonin-based targetable fluorescent probe activated by hydrogen peroxide for tumor cells screening

2021 ◽  
pp. 131051
Author(s):  
Xiwei Li ◽  
Na Gao ◽  
Caiyun Liu ◽  
Miaohui Yu ◽  
Xiaodi Rong ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Tumor Cells ◽  
Fluorescent Probe

The Effect of Sirtuin 2 (Sirt2) Overexpressing Bone Marrow Mesenchymal Stem Cells on the Growth of Human Epidermal Growth Factor Receptor 2 (Her-2) Breast Cancer Cells and Its Mechanism

2021 ◽  
Vol 11 (4) ◽  
pp. 778-785
Author(s):  
Xiaolin Chen ◽  
Yan Wang ◽  
Sunlu Jiang
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Nk Cells ◽  
Tumor Cells ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Caspase 3 ◽  
Tumor Bearing ◽  
Her 2 ◽  
Ifn Γ

Our study investigates the effect of high expression of Sirt2 in MSCs (MSCs-Sirt2) on Her-2 breast cancer cell proliferation. A mouse subcutaneous xenograft tumor model was established and MSCssirt2 analysis was performed on nude mice. TUNEL staining, flow cytometry, western-blot, real-time PCR and immunohistochemistry were used to detect cancer cell apoptosis. The number of NK cells infiltrated by flow cytometry detected the tumor tissue of tumor-bearing mice, and its killing activity on tumor-bearing mice was detected by isotope labeling and release method. The levels of TNF-α, IFN-γ, IL-8, IL-6 and IL-10 were detected by ELISA. Caspase-3 level was decreased in the MSCs group (P <0.01) while increased in the MSCs-sirt2 group (P <0.001). However, PCNA expression showed an opposite profile in the Her-2 group and MSCs-sirt2 group compared to Caspase-3 level (P <0.01). The tumor volume and weight in the MSCs-sirt2 group was significantly reduced (P < 0.01), while increased in the MSCs group significantly (P < 0.05). The number of Ki-67-positive tumor cells in MSCs-sirt2 group was significantly reduced (P <0.01) and increased in MSCs group (P < 0.001) with oppositive number of TUNEL-positive tumor cells in the MSCs-sirt2 group and MSCs group (P <0.01). IFN-γ level showed an upward trend (P <0.001). The NK cell toxicity of MSCs-Sirt2 group was significantly higher (P <0.001). MSCs-Sirt2 has an inhibitory effect on Her-2 breast cancer cell growth by enhancing the local inflammatory response of NK cells.

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