Application of the BrdU/thymidine method to flow cytogenetics: Differential quenching/enhancement of Hoechst 33258 fluorescence of late-replicating chromosomes

1982 ◽  
Vol 8 (3) ◽  
pp. 319-327 ◽  
Author(s):  
C. Cremer ◽  
J. W. Gray
Keyword(s):  
Hoechst 33258 ◽  
Differential Quenching

Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability

Reproduction ◽  
2000 ◽  
pp. 145-152 ◽  
Author(s):  
B Pintado ◽  
J de la Fuente ◽  
ER Roldan
Keyword(s):  
Nucleic Acid ◽  
Propidium Iodide ◽  
Hoechst 33258 ◽  
Experimental Conditions ◽  
Functional Studies ◽  
Viable Cells ◽  
Direct Microscopic Examination ◽  
Acrosomal Exocytosis ◽  
Viable Sperm ◽  
Boar Spermatozoa

This study was designed to assess whether nucleic acid stains such as propidium iodide and Hoechst 33258 and the cytosolic stain eosin identified equivalent proportions of non-viable cells. Sub-samples of boar spermatozoa stored for up to 72 h, and frozen bull spermatozoa stored in straws and thawed before staining, were exposed to either propidium iodide or Hoechst 33258 alone or in combination. Additional sub-samples were stained with eosin-nigrosin and subsequently with Giemsa. The proportion of non-viable cells identified by propidium iodide alone was equivalent to that observed when it was used in combination with the other fluorescent probe. Similar results were observed for Hoechst 33258. However, direct microscopic examination of sub-samples exposed to both stains revealed that a proportion of spermatozoa stained with propidium iodide did not incorporate Hoechst 33258. This was found consistently in boar and bull spermatozoa under the different experimental conditions used. Quantification showed that the proportion of propidium iodide-positive cells was significantly higher than Hoechst 33258-positive cells. Furthermore, the proportion of propidium iodide-positive cells was higher than cells stained with eosin, but no differences were found between the number of cells stained with Hoechst 33258 or eosin. The proportion of cells stained with propidium iodide was positively correlated with the proportion stained with either Hoechst 33258 or eosin, despite the observation that more cells incorporated propidium iodide. Taken together, these results indicate that there are differences in the ability of fluorescent probes to identify non-viable sperm cells and that this should be considered when staining protocols are used to analyse sperm viability, or when viability is used as a discriminating factor in functional studies, such as those related to acrosomal exocytosis.

DNA electrochemical biosensor for detection of Alicyclobacillus acidoterrestris utilizing Hoechst 33258 as indicator

2021 ◽  
pp. 107801
Author(s):  
José M. R. Flauzino ◽  
Rafaela C. S. Peres ◽  
Lívia M. Alves ◽  
Jussara G. Vieira ◽  
Júlia G. dos Santos ◽  
...  
Keyword(s):  
Electrochemical Biosensor ◽  
Hoechst 33258 ◽  
Alicyclobacillus Acidoterrestris ◽  
Dna Electrochemical Biosensor

Binding of Hoechst 33258 to the minor groove of B-DNA

1987 ◽  
Vol 197 (2) ◽  
pp. 257-271 ◽  
Author(s):  
Philip E. Pjura ◽  
Kazimierz Grzeskowiak ◽  
Richard E. Dickerson
Keyword(s):  
Minor Groove ◽  
Hoechst 33258

STUDY OF ABSORPTION SPECTRA OF THE COMPLEXES OF BOVINE SERUM ALBUMIN WITH HOECHST 33258 AND METHYLENE BLUE

2021 ◽  
Vol 55 (2 (255)) ◽  
pp. 158-164
Author(s):  
Nara H. Petrosyan
Keyword(s):  
Methylene Blue ◽  
Serum Albumin ◽  
Absorption Spectra ◽  
Hoechst 33258 ◽  
Differential Absorption ◽  
Partial Loss ◽  
Groove Binding ◽  
Compact Structure ◽  
Binding Material ◽  
Molecular Compounds

The study on the interaction of DNA-specific low-molecular compounds – groove binding material Hoechst 33258 and intercalating ligand methylene blue (MB) with serum albumin has been carried out. The absorption and differential absorption spectra of complexes of the mentioned ligands with protein were obtained. Changes of the absorption and differential absorption spectra indicate the binding of two ligands with albumin. The obtained results indicate that at the interaction with both ligands, the conformational state of the protein alters, though these changes are not similar, since in the case of MB a compactization of the protein folding occurs, while in the case of Hoechst 33258, most apparently, an unfolding of the compact structure takes place as a result of partial loss of helicity of $\alpha$-structures.

Condensation of the chromatin at the membrane of an apoptotic nucleus is not associated with activation of an endonuclease

1993 ◽  
Vol 104 (2) ◽  
pp. 317-326 ◽  
Author(s):  
F. Oberhammer ◽  
G. Fritsch ◽  
M. Schmied ◽  
M. Pavelka ◽  
D. Printz ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Early Event ◽  
Tgf Beta ◽  
Hoechst 33258 ◽  
Nucleolar Structure ◽  
Nuclear Changes ◽  
Rigorous Test ◽  
A Current ◽  
Mouse Lymphoma

A current hypothesis holds that chromatin fragmentation into oligonucleosomal patterns is an early event during apoptosis. In contrast, induction of apoptosis in cultured hepatocytes by TGF-beta 1 was not associated with DNA fragmentation into oligonucleosomes in hepatocyte monolayers and apoptotic fragments. For a more rigorous test of the hypothesis we performed a number of experiments. We compared nuclear changes resulting from TGF-beta 1 with those induced by Ca2+, a known activator of endonuclease. The morphology of apoptotic and Ca(2+)-treated nuclei was different as judged by DNA staining with Hoechst 33258. Likewise, electron microscopy of apoptotic nuclei showed characteristic condensation of the chromatin as well as dissolution of the nucleolar structure and nuclear fragmentation, changes not seen after Ca2+ treatment, after three hours of incubation. Analysis of DNA fluorescence of nuclei by FACS revealed that treatment with Ca2+ reduced the signal by 20%. In contrast, nuclei from TGF-beta 1-treated hepatocytes did not exhibit a reduced signal and after sorting by FACS, apoptotic nuclei remained in the 2N and 4N fractions. The absence of detectable DNA fragmentation in apoptotic nuclei was further verified by in situ nick translation, not only in hepatocytes but also in a mouse lymphoma cell line. From these findings we conclude that activation of an endonuclease is not an early event on the pathway to morphologically recognizable apoptosis.

Exploring the Interaction of Ethidium Bromide and HOECHST 33258 with DNA by Means of Electrochemical Approach

2017 ◽  
Vol 12 (03) ◽  
pp. 151-161
Author(s):  
Levon A. Tavadyan ◽  
Seyran H. Minasyan ◽  
Gaspar H. Kocharyan ◽  
Ara P. Antonyan ◽  
Viktoria G. Sahakyan ◽  
...  
Keyword(s):  
Ethidium Bromide ◽  
Square Wave Voltammetry ◽  
Differential Pulse ◽  
Binding Modes ◽  
Hoechst 33258 ◽  
Wave Voltammetry ◽  
Calf Thymus ◽  
Electrochemical Approach ◽  
Pulse Voltammetry ◽  
Binding Type

The binding of both ethidium bromide (EtBr) and Hoechst 33258 (H33258) to calf thymus DNA differing by the mechanism at different ionic strengths of NaCl water solutions (2, 20 and 150[Formula: see text]mM) has been quantitatively studied by square wave voltammetry (SWV). It was revealed that EtBr binds to DNA by more than one mode: at the solution ionic strengths 2 and 150[Formula: see text]mM, two modes were revealed, at 20[Formula: see text]mM — three modes. Values of EtBr binding constant ([Formula: see text]) to DNA and the number of nucleotides per one binding site ([Formula: see text]) for respective binding modes were determined. For H33258 under the same conditions, two binding modes were found and for both these modes values of [Formula: see text] and [Formula: see text] were determined. Dependence of the binding types and parameters on the solution ionic strength by SWV method was revealed, which differs by the binding type of EtBr and H33258. The obtained data are in good accordance with the ones determined by spectroscopic (absorption and fluorescence) and differential pulse voltammetry (DPV) methods. The obtained data indicate the sensibility and high precision of SWV method which may be applied to studies on interaction of different ligands with DNA, which reveal some peculiarities of their binding to DNA that are hidden and are not found by other methods.

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