We present a simple and robust method to evaluate the transfection efficiency of commercially available transfection reagents intended to be established for use in nonmammalian cell lines. To illustrate the method, we compare the ability of four different reagents to transfect the embryonic zebrafish cell line Z3. Z3 cells were seeded in a 96-well plate and simultaneously transfected in several...

A small stem-like subpopulation was isolated from human hepatocellular carcinoma (HCC) MHCC97 cells, characterized by their high efflux ability of the Hoechst 33342 dye. These side population (SP) cells were able to generate a heterogeneous mixture of SP and main population (MP) cells, while the MP cells rarely generated SP cells. Cell cycle analysis also revealed that more SP cells were in the...

Nowadays, chemotherapy is still the main effective treatment for cancer. Herb prescriptions containing Pogostemon cablin Benth (also known as "Guang-Huo-Xiang") have been widely used in Chinese medicine today. In our research, we found that patchouli alcohol, a compound isolated from the oil of Pogostemon cablin Benth, exerted antitumor ability against human lung cancer A549 cells ability both ...

Material and Methods Materials. Neurobasal medium, B27, SUPERSCRIPT II RNase H reverse transcriptase, RNAaseOUT, and Taq polymerase were from Invitrogen (Paisley, Scotland); papain was from Worthington Biochemical Corp. (Lakewood, NJ); NaAsO2, cysteine, soybean trypsin inhibitor (STI), propidium iodide (PI), Hoechst 33342, and the bicinchoninic acid protein assay kit were from Sigma (Poole, Dor...

After a rapid overview of the approaches used to study cell cycle, a fluorescent digital imaging microscopy method is proposed. This method is improved by a factorial analysis relying on the evaluation of several parameters recorded on each living cell. Single lympho-blastoid living cells are labeled with three fluorescent markers: Hoechst 33342 for nuclear DNA, Rhodamine 123 for mitochondria a...

The relationship between the concentration of cell-free DNA (cfDNA) and the number of proliferating/apoptotic lymphocytes in peripheral blood of premature newborns of different gestation age and full-term newborns was determined. The experiments were performed using fluorescent spectrophotometry (with Hoechst 33342), flow cytometry, and microscopy (Feulgen staining of lymphocytes). It was deter...

The combination of multiple dye-DNA interactions, a fluorescence digital imaging system with a scientific CCD camera, and multivariate image analysis allows the rapid karyotyping of fluorescent human metaphase chromosome spreads. Chromosomes are stained with the bisbenzimidazole dye Hoechst 33342 and chromomycin A3, a dye pair used frequently in bivariate flow analysis and sorting of metaphase ...

The MTT assay, as a screening method, has been widely used to measure the viability and proliferation of cells. However, it should be noted that MTT assay may not accurately reflect the effect of Cistanche tubulosa ethanolic extract on EA.hy926 cells viability. To investigate and identity the components responsible for the contradictory observations of the MTT assay, echinacoside and acteoside,...

STK295900, a small synthetic molecule belonging to a class of symmetric bibenzimidazoles, exhibits antiproliferative activity against various human cancer cell lines from different origins. Examining the effect of STK295900 in HeLa cells indicates that it induces G(2) phase arrest without invoking DNA damage. Further analysis shows that STK295900 inhibits DNA relaxation that is mediated by topo...

Due to the presence and activity of mammary stem cells (MaSC), growth and remodelling of mammary gland during puberty and lactation cycles is possible. In this study the number of putative mammary stem/progenitor cells was examined in 20 months old non-pregnant Holstein-Friesian heifers. Cells were double-stained with fluorescent dye-conjugated antibodies against stem cell antigen-1 (Sca-1) and...