Use of fluorescent staining and flow cytometry for monitoring physiological changes in solventogenic clostridia

Anaerobe ◽  
2014 ◽  
Vol 29 ◽  
pp. 113-117 ◽  
Author(s):  
Petra Patakova ◽  
Michaela Linhova ◽  
Pavla Vykydalova ◽  
Barbora Branska ◽  
Mojmir Rychtera ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Physiological Changes ◽  
Fluorescent Staining ◽  
Solventogenic Clostridia

scholarly journals Nilotinib, a Discoidin domain receptor 1 (DDR1) inhibitor, induces apoptosis and inhibits migration in breast cancer

2019 ◽  
Author(s):  
Shufang Yu ◽  
Shuang Wang ◽  
Yanping Xie ◽  
Aina Bao ◽  
Jing Li ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Cell Death ◽  
Apoptotic Cell ◽  
Cellular Migration ◽  
Fluorescent Staining ◽  
Discoidin Domain Receptor ◽  
Apoptotic Activity ◽  
Discoidin Domain Receptor 1 ◽  
Mcf 7

Abstract Background: Overexpression of Discoidin domain receptor 1(DDR1) is known to enhance the malignancy of breast cancer considerably. This study reports the identification of a potent DDR1 inhibitor, Nilotinib for the treatment of breast cancer. Methods: MTT assay was used to evaluate the inhibitory activity of Nilotinib and meantime we used flow cytometry to evaluate the pro-apoptotic activity of Nilotinib against MCF-7 and MDA-MB-231. Expression of DDR1 was manipulated in MCF-7 cell lines with low level DDR1 expression by transfecting with plasmids containing by shRNA. The effect of DDR1 or treatment with Nilotinib on cell migration was assayed. The expression of p-DDR1, DDR1, p-ERK1/2, ERK1/2 and E-cadherin，Vimentin, Snail1 were detected by western blot and immuno-fluorescent staining . Results: Nilotinib against MCF-7 (IC 50 =0.403 μM) and MDA-MB-231 (IC 50 =0.819 μM) and also indicated induced apoptotic cell death after co-cuturing with Nilotinib (500 nM), apoptosis rate is 29.60 % ± 2.19% and 18.75 % ± 2.30% respective ly. Moreover, Nilotinib effectually blocked cellular migration of MCF-7. Interestingly, Knock-down DDR1 could significantly block the migration of breast cancer, meantime the sensitivity of Nilotinib to MCF-7 was reduced. Conclusion: Targeting DDR1 therapeutically could potentially affect survival and influence metabolism in breast cancer, and Nilotinib could as a candidate for the treatment of breast cancer.

Post-thaw evaluation of dog spermatozoa using new triple fluorescent staining and flow cytometry

1999 ◽  
Vol 52 (6) ◽  
pp. 965-980 ◽  
Author(s):  
A. Peña ◽  
A. Johannisson ◽  
Linde-Forsberg
Keyword(s):  
Flow Cytometry ◽  
Fluorescent Staining

scholarly journals Viability Assessment of Turkey Sperm Using Fluorescent Staining and Flow Cytometry

1995 ◽  
Vol 74 (7) ◽  
pp. 1191-1200 ◽  
Author(s):  
ANN M. DONOGHUE ◽  
DUANE L. GARNER ◽  
DAN J. DONOGHUE ◽  
LAWRENCE A. JOHNSON
Keyword(s):  
Flow Cytometry ◽  
Fluorescent Staining ◽  
Viability Assessment

scholarly journals Rapid assessment of antibiotic effects on Escherichia coli by bis-(1,3-dibutylbarbituric acid) trimethine oxonol and flow cytometry.

1997 ◽  
Vol 41 (9) ◽  
pp. 2001-2005 ◽  
Author(s):  
R I Jepras ◽  
F E Paul ◽  
S C Pearson ◽  
M J Wilkinson
Keyword(s):  
Escherichia Coli ◽  
Flow Cytometry ◽  
Membrane Potential ◽  
Rapid Assessment ◽  
Physiological Changes ◽  
Analytical Technique ◽  
E Coli ◽  
Flow Cytometric ◽  
Bacterial Cell Viability ◽  
Sensitive Analytical Technique

The effects of selected antibiotics on Escherichia coli were studied by flow cytometry with the fluorescent anionic membrane potential probe bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)]. The actions of azithromycin, cefuroxime, and ciprofloxacin at five times the MIC on E. coli were compared by the traditional CFU assay and flow cytometry. Changes in viable counts of bacteria determined with DiBAC4(3) and by flow cytometry following treatment with the antibiotics showed trends similar to those found by the CFU assays. However, viable counts determined by flow cytometry following antibiotic treatment were 1 to 2 logs higher than those determined by the corresponding CFU assays. All the results obtained by flow cytometry were provided within 10 min after sampling, whereas the conventional CFU assay results took at least 18 h. The results indicated that flow cytometry is a sensitive analytical technique that can rapidly monitor the physiological changes of individual microorganisms following antibiotic action and can provide information on the mode of action of a drug. The membrane potential probe DiBAC4(3) provides a robust flow cytometric indicator for bacterial cell viability.

scholarly journals Nilotinib, a Discoidin domain receptor 1 (DDR1) inhibitor, induces apoptosis and inhibits migration in breast cancer

2020 ◽  
Author(s):  
Shufang Yu ◽  
Shuang Wang ◽  
Yanping Xie ◽  
Aina Bao ◽  
Jing Li ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Cell Death ◽  
Apoptotic Cell ◽  
Cellular Migration ◽  
Fluorescent Staining ◽  
Discoidin Domain Receptor ◽  
Apoptotic Activity ◽  
Discoidin Domain Receptor 1 ◽  
Mcf 7

Abstract Background: Overexpression of Discoidin domain receptor 1(DDR1) is known to enhance the malignancy of breast cancer considerably. This study reports the identification of a potent DDR1 inhibitor, Nilotinib for the treatment of breast cancer. Methods: MTT assay was used to evaluate the inhibitory activity of Nilotinib and meantime we used flow cytometry to evaluate the pro-apoptotic activity of Nilotinib against MCF-7 and MDA-MB-231. Expression of DDR1 was manipulated in MCF-7 cell lines with low level DDR1 expression by transfecting with plasmids containing by shRNA. The effect of DDR1 or treatment with Nilotinib on cell migration was assayed. The expression of p-DDR1, DDR1, p-ERK1/2, ERK1/2 and E-cadherin，Vimentin, Snail1 were detected by western blot and immuno-fluorescent staining . Results: Nilotinib against MCF-7 (IC 50 =0.403 μM) and MDA-MB-231 (IC 50 =0.819 μM) and also indicated induced apoptotic cell death after co-cuturing with Nilotinib (500 nM), apoptosis rate is 29.60 % ± 2.19% and 18.75 % ± 2.30% respective ly. Moreover, Nilotinib effectually blocked cellular migration of MCF-7. Interestingly, Knock-down DDR1 could significantly block the migration of breast cancer, meantime the sensitivity of Nilotinib to MCF-7 was reduced. Conclusion: Targeting DDR1 therapeutically could potentially affect survival and influence metabolism in breast cancer, and Nilotinib could as a candidate for the treatment of breast cancer.

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