Screening for Anticancer Activity: Dual Staining Method

Author(s):  
Ramachandran Chelliah ◽  
Deog-Hwan Oh
2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Hai Trieu Ly ◽  
Trieu Minh Truong ◽  
Thi Thu Huong Nguyen ◽  
Hoang Dung Nguyen ◽  
Yuxia Zhao ◽  
...  

Abstract Background Cancer is one of the most considerable concerns because of increasing the death rate all over the world. Recent studies have disclosed that plant extracts exhibit anticancer activity through various mechanisms. Xanthium strumarium has been used by Vietnamese in herbal medicines to support the medication of infirmities. This study is to consider the secondary metabolites, antioxidant and anticancer capacities of extract from the aerial parts (stems and leaves) of X. strumarium (AP-XS). Methods AP-XS was analyzed for the presence of phytochemicals via qualitative chemical tests and determined total polyphenol and flavonoid contents. DPPH (1,1-diphenyl-2-picrylhydrazyl) quenching assay and sulforhodamine B (SRB) assay were selected to investigate antioxidant capacity and anti-proliferative activity, respectively. Besides, acridine orange-ethidium bromide (AO-EB) dual staining was applied to evaluate the ability to induce apoptosis on HepG2 cancer cells. Results Results of present study indicated that AP-XS contains the main phytochemicals such as flavonoids, tannins, saponins, alkaloids, and triterpenes. Ethanol extract had highest content of polyphenol (84.86 mg gallic acid equivalent/g dry mass), and exhibited the great total antioxidant property (IC50 = 184.13 μg/mL) and anti-proliferative activity on HepG2 cancer cells (IC50 = 81.69 μg/mL). Furthermore, the characteristics of apoptosis including shrinkage of the cell and apoptotic bodies were found following 60 h of AP-XS extract treatment through AO-EB dual staining. Conclusion The data suggest that AP-XS extract had antioxidant potential and anti-proliferative effect. The anti-proliferative property was considered to have an association with a rising of apoptosis. These results were reliable for further research on X. strumarium as a source of phytochemicals with anticancer activity potential for cancer therapeutics.


1991 ◽  
Vol 23 (1) ◽  
pp. 22-28 ◽  
Author(s):  
H. Ota ◽  
T. Katsuyama ◽  
K. Ishii ◽  
J. Nakayama ◽  
T. Shiozawa ◽  
...  

1975 ◽  
Vol 2 (2) ◽  
pp. 149-150
Author(s):  
N. M. Burdash ◽  
M. E. West ◽  
E. R. Bannister ◽  
C. Dyar ◽  
R. C. Duncan

A dual-staining procedure for acid-fast bacilli was found to have poor correlation with the Ziehl-Neelsen and auramine-rhodamine staining techniques.


2000 ◽  
Vol 20 (4) ◽  
pp. 151-154 ◽  
Author(s):  
Lukas Günther ◽  
Peter Hufnagl

A new staining method for dual demonstration of Estrogen receptors (ER) and argyrophilc Nucleolus‐Organizer Regions (AgNORs) was developed. To rule out possible reciprocal effects, serial slides of 10 invasive ductale breast cancers were stained with either the single staining method or the simultaneous ER/AgNOR‐staining method and investigated comparatively. By measuring the slides with the image analysis system AMBA, reciprocal effects could be excluded. It was proven that dual staining of both markers results in a reproducible and specific staining result. We concluded that it is justified to measure AgNORs in immunohistochemically stained cells.


2007 ◽  
Vol 128 (3) ◽  
pp. 275-283 ◽  
Author(s):  
Douglas J. Taatjes ◽  
Marilyn P. Wadsworth ◽  
A. K. M. Tarikuz Zaman ◽  
David J. Schneider ◽  
Burton E. Sobel

Author(s):  
E. N. Albert

Silver tetraphenylporphine sulfonate (Ag-TPPS) was synthesized in this laboratory and used as an electron dense stain for elastic tissue (Fig 1). The procedures for the synthesis of tetraphenylporphine sulfonate and the staining method for mature elastic tissue have been described previously.The fine structure of developing elastic tissue was observed in fetal and new born rat aorta using tetraphenylporphine sulfonate, phosphotungstic acid, uranyl acetate and lead citrate. The newly forming elastica consisted of two morphologically distinct components. These were a central amorphous and a peripheral fibrous. The ratio of the central amorphous and the peripheral fibrillar portion changed in favor of the former with increasing age.It was also observed that the staining properties of the two components were entirely different. The peripheral fibrous component stained with uranyl acetate and/or lead citrate while the central amorphous portion demonstrated no affinity for these stains. On the other hand, the central amorphous portion of developing elastic fibers stained vigorously with silver tetraphenylporphine sulfonate, while the fibrillar part did not (compare figs 2, 3, 4). Based upon the above observations it is proposed that developing elastica consists of two components that are morphologically and chemically different.


Author(s):  
K. Chien ◽  
I.P. Shintaku ◽  
A.F. Sassoon ◽  
R.L. Van de Velde ◽  
R. Heusser

Identification of cellular phenotype by cell surface antigens in conjunction with ultrastructural analysis of cellular morphology can be a useful tool in the study of biologic processes as well as in diagnostic histopathology. In this abstract, we describe a simple pre-embedding, protein A-gold staining method which is designed for cell suspensions combining the handling convenience of slide-mounted cell monolayers and the ability to evaluate specimen staining specificity prior to EM embedding.


Author(s):  
Anne M. Klinkner ◽  
Crystal R. Waites ◽  
Peter J. Bugelski ◽  
William D. Kerns

A primary effort in the understanding of the progression of atherosclerotic disease has been methods development for visualization of the atherosclerotic plaque. We introduce a new method for the qualitative analysis of lipids in atherosclerotic fatty streaks which also retains those lipids for biochemical evaluation. An original aspect of the process is the ability to view an entire fatty streak en face, selectively stained for specific lipid classes within the lesion.New Zealand white rabbits were fed a high cholesterol diet(0.15%-0.3% for 14 wks). The aorta was removed and fixed in Carson's phosphate buffered formaldehyde followed by dual staining in the fluorescent dyes Nile red and filipin. Stock solutions of nile red(0.5mg/ml acetone) and filipin(2.5mg/ml dimethyl formamide) were prepared and kept at -20°C; all subsequent steps were at RT. 0.5cm × 1.0cm pieces of aorta were trimmed and adventitia removed. The pieces were then washed 3×15 min in PBS w/o CaMg, soaked in Nile red(NR)/filipin(Fl) stain(100(il NR stock + 200μl Fl stock in 10 ml PBS for 30 min, washed in PBS 3×30 min, rinsed with distilled water, mounted(Crystal Mount, Biomedia) and coverslipped and viewed by fluorescence microscopy.


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