Automated determination of DNA using the fluorochrome Hoechst 33258

Sterzel, W.; Bedford, Philip; Eisenbrand, Gerhard

doi:10.1016/0003-2697(85)90299-4

Cited by 77 publications

(31 citation statements)

References 9 publications

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“…Hoechst dyes are commonly used to stain genomic DNA in applications such as fluorescence microscopy and immunohistochemistry, often together with other fluorophores (Johnson & Michelle, 2011), flow cytometry to count or sort out cells, detection of DNA in the presence of RNA in agarose gels (Mocharla et al, 1987), automated DNA determination and chromosome sorting (Sterzel et al, 1985).…”

Section: Hoechst 33342 Stainingmentioning

confidence: 99%

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Atale

Gupta

Yadav³

et al. 2014

Journal of Microscopy

278

171

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Summary Apoptosis, a genetically programmed cellular event leads to biochemical and morphological changes in cells. Alterations in DNA caused by several factors affect nucleus and ultimately the entire cell leading to compromised function of the organ and organism. DNA, a master regulator of the cellular events, is an important biomolecule with regards to cell growth, cell death, cell migration and cell differentiation. It is therefore imperative to develop the staining techniques that may lead to visualize the changes in nucleus where DNA is housed, to comprehend the cellular pathophysiology. Over the years a number of nuclear staining techniques such as propidium iodide, Hoechst‐33342, 4’, 6‐diamidino‐2‐phenylindole (DAPI), Acridine orange–Ethidium bromide staining, among others have been developed to assess the changes in DNA. Some nonnuclear staining techniques such as Annexin‐V staining, which although does not stain DNA, but helps to identify the events that result from DNA alteration and leads to initiation of apoptotic cell death. In this review, we have briefly discussed some of the most commonly used fluorescent and nonfluorescent staining techniques that identify apoptotic changes in cell, DNA and the nucleus. These techniques help in differentiating several cellular and nuclear phenotypes that result from DNA damage and have been identified as specific to necrosis or early and late apoptosis as well as scores of other nuclear deformities occurring inside the cells.

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Section: Hoechst 33342 Stainingmentioning

confidence: 99%

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Atale

Gupta

Yadav³

et al. 2014

Journal of Microscopy

278

171

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“…It was monitored by the formation of malondialdehyde (Smith et al, 1 9 8 2 ) . DNA content was determined fluorometrically according to Sterzel et al (1985).…”

Section: Other Proceduresmentioning

confidence: 99%

Mechanism of Carbon Tetrachloride-Induced Hepatotoxicity. Hepatocellular Damage by Reactive Carbon Tetrachloride Metabolites

Boll

Weber

Becker

et al. 2001

Zeitschrift Für Naturforschung C

257

140

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Liver Damage, Carbon Tetrachloride, PeroxidationCCl4-induced liver damage was modeled in monolayer cultures of rat primary hepatocytes with a focus on involvement of covalent binding of CC14 metabolites to cell components and/ or peroxidative damage as the cause of injury.(1) Covalent binding of 1 4 C-labeled metabolites was detected in hepatocytes immediately after exposure to CC14. (2) Low oxygen partial pressure increased the reductive metabolism of CC14 and thus covalent binding. (3) [1 4 C]-CC14 was bound to lipids and to proteins throughout subcellular fractions. Binding occurred preferentially to triacylglycerols and phos pholipids, with phosphatidylcholine containing the highest amount of label. (4) The lipid peroxidation potency of CC14 revealed subtle differences compared to other peroxidative substances, viz., A D P-Fe3+ and cumol hydroperoxide, respectively. (5) CC14, but not the other peroxidative substances, decreased the rate of triacylglycerol secretion as very low density lipoproteins. (6 ) The anti-oxidant vitamin E (a-tocopherol) blocked lipid peroxidation, but not covalent binding, and secretion of lipoproteins remained inhibited. (7) The radical scav enger piperonyl butoxide prevented CCl4-induced lipid peroxidation as well as covalent bind ing of CCI4 metabolites to cell components, and also restored lipoprotein metabolism.The results confirm that covalent binding of the CC13* radical to cell components initiates the inhibition of lipoprotein secretion and thus steatosis, whereas reaction with oxygen, to form CCI3 -O O * , initiates lipid peroxidation. The two processes are independent of each other, and the extent to which either process occurs depends on partial oxygen pressure. The former process may result in adduct formation and, ultimately, cancer initiation, whereas the latter results in loss of calcium homeostasis and, ultimately, apoptosis and cell death.

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“…Other methods DNA was determined by a fluorometric method using the Hoechst 33258 dye, as described by Sterzel et al (1985). Protein was measured by the method of Lowry et al (1951).…”

Section: Estimation Of Dna Synthesismentioning

confidence: 99%

Stimulation of hepatocyte DNA synthesis by prostaglandin E₂ and prostaglandin F₂α additivity with the effect of norepinephrine, and synergism with epidermal growth factor

Refsnes

et al. 1994

Journal Cellular Physiology

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Previous data obtained in vivo and in vitro suggest that both prostaglandins (PGs) and catecholamines may have a role in promoting hepatocyte proliferation, and PGE2 and PGF2 alpha have also been implicated as mediators of the mitogenic actions of epidermal growth factor (EGF) (and transforming growth factor alpha [TGF alpha]). We have studied the effects of PGs and norepinephrine on DNA synthesis in serum-free primary cultures of rat hepatocytes, and compared the PG effects with those of norepinephrine. PGE2, PGF2 alpha, PGD2, and the synthetic analog dimethyl-PGE2 markedly enhanced the DNA synthesis. A more quantitative analysis of the effects of PGE2 and PGF2 alpha on the DNA synthesis, in the presence and absence of EGF, indicated that these PGs interacted in an essentially multiplicative manner with the effect of EGF. The effects of PGE2 and PGF2 alpha showed almost complete additivity with the stimulation of DNA synthesis produced by maximally effective concentrations of norepinephrine. The data suggest a) that PGE2 and PGF2 alpha facilitate and synergize with, rather than mediate, the actions of EGF in hepatocytes, and b) that this effect of the PGs occurs by mechanisms that are at least partly distinct from those of norepinephrine.

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Automated determination of DNA using the fluorochrome Hoechst 33258

Cited by 77 publications

References 9 publications

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Mechanism of Carbon Tetrachloride-Induced Hepatotoxicity. Hepatocellular Damage by Reactive Carbon Tetrachloride Metabolites

Stimulation of hepatocyte DNA synthesis by prostaglandin E₂ and prostaglandin F₂α additivity with the effect of norepinephrine, and synergism with epidermal growth factor

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Automated determination of DNA using the fluorochrome Hoechst 33258

Cited by 77 publications

References 9 publications

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Cell‐death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques

Mechanism of Carbon Tetrachloride-Induced Hepatotoxicity. Hepatocellular Damage by Reactive Carbon Tetrachloride Metabolites

Stimulation of hepatocyte DNA synthesis by prostaglandin E2 and prostaglandin F2α additivity with the effect of norepinephrine, and synergism with epidermal growth factor

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Stimulation of hepatocyte DNA synthesis by prostaglandin E₂ and prostaglandin F₂α additivity with the effect of norepinephrine, and synergism with epidermal growth factor