scholarly journals STUDIES ON NUCLEIC ACID METACHROMASY

1965 ◽  
Vol 27 (2) ◽  
pp. 327-336 ◽  
Author(s):  
Ned Feder ◽  
Merrill K. Wolf
Keyword(s):  
Nucleic Acid ◽  
Ribosomal Rna ◽  
Toluidine Blue ◽  
Color Contrast ◽  
Tissue Sections ◽  
Dna And Rna ◽  
Excellent Preservation ◽  
Structural Preservation ◽  
Intense Color

Acrolein-fixed, polyester wax-embedded tissue sections showed excellent preservation of light microscopic architecture and, when stained with toluidine blue, intense color contrast between DNA, which stained orthochromatically, and RNA, which stained metachromatically. This method has practical value for differentiating DNA from RNA in the same section. The color contrast was impaired by substituting formaldehyde for acrolein or paraffin for polyester wax, and was negligible in tissues fixed in formaldehyde or Carnoy's fluid and embedded in paraffin. Quality of structural preservation paralleled degree of color contrast. Metachromatic staining can be analysed, by the quantitative parameters of Bradley and colleagues, to provide inferences regarding the conformation of biopolymers in tissue sections. Comparison of the nucleic acid color contrasts in toluidine blue-stained sections with titrations of fixative-treated nucleic acids against toluidine blue in solution indicated a greater difference in conformation between DNA- and RNA-protein in acrolein-polyester sections than between acrolein-treated free DNA and RNA in solution. This is supported by recent evidence that the conformation of ribosomal RNA is quite different in whole ribosomes from that assumed by the same RNA free in solution. The acrolein-polyester method may enhance color contrast by providing superior preservation of ordered nucleoprotein conformations.

scholarly journals STUDIES ON NUCLEIC ACID METACHROMASY

1965 ◽  
Vol 27 (2) ◽  
pp. 313-326 ◽  
Author(s):  
Michael E. Lamm ◽  
Lillian Childers ◽  
Merrill K. Wolf
Keyword(s):  
Nucleic Acid ◽  
Toluidine Blue ◽  
Binding Sites ◽  
Heat Denaturation ◽  
Color Contrast ◽  
Fluid Absorption ◽  
Tissue Sections ◽  
Dna And Rna ◽  
Nucleic Acid Conformation ◽  
Intense Color

The stacking coefficients (K's) of nucleic acids have been thought to influence the color contrast between DNA and RNA in tissue sections stained with metachromatic dyes. This idea was tested by titrating toluidine blue (TB) and acridine orange (AO) in solution against DNA and RNA, native or treated with formaldehyde, acrolein, or Carnoy's fluid. Absorption spectra at varying polymer-dye ratios were used to compute K values by the methods of Bradley and colleagues. Results with both dyes fit Bradley's stacking equations. Fixatives did not block dye-binding sites but markedly altered K values. K of DNA was low, unaffected by aldehyde fixative, increased by Carnoy's fluid or heat denaturation. K of RNA was higher than that of DNA and was increased greatly by formaldehyde, almost as much by acrolein, considerably less by Carnoy's fluid. Aldehyde effects were partially reversed upon removal of aldehyde by dialysis. These observations accord with known effects of aldehydes and denaturation upon nucleic acid conformation. Differences between K's of DNA and RNA were greater after aldehyde treatment than after Carnoy's, and were greater with AO than with TB. This is generally consistent with the magnitude of the color contrasts observed in tissues. Additional factors must contribute to the intense color contrast observed in acrolein-fixed tissues stained with TB.

Morphological quality and nucleic acid preservation in cytopathology

2008 ◽  
Vol 62 (5) ◽  
pp. 429-434 ◽  
Author(s):  
A Gazziero ◽  
V Guzzardo ◽  
E Aldighieri ◽  
A Fassina
Keyword(s):  
Nucleic Acid ◽  
Molecular Mass ◽  
Molecular Analysis ◽  
Molecular Techniques ◽  
Fine Needle ◽  
Rna Integrity ◽  
Fine Needle Aspirates ◽  
Dna And Rna ◽  
Excellent Preservation ◽  
Formalin Fixed

Background:Fixation is a chemical or physical procedure to prevent the degradation of proteins and tissue morphology. To optimise molecular analysis of archival tissues, it is essential that fixation preserves morphology along with protein epitopes and DNA/RNA integrity.Methods:A new formalin-free alcoholic-based fixative, FineFIX, was used to fix 15 serous effusions and 38 fine-needle aspirates, and cellular morphology and nucleic acid quality were evaluated.Results:The cytomorphology of the effusions and fine-needle aspirates obtained with FineFIX fixation was similar to that obtained with formalin-fixed counterparts. Immunocytochemistry showed comparable results with the traditional fixative, but FineFIX preserved higher-molecular-mass DNA and RNA, as demonstrated by successful PCR of large amplification products of >2000 bp.Conclusions:The formalin-free fixative produced not only satisfactory results for immunocytochemistry on cytological smears and cell blocks, but also excellent preservation of DNA and RNA, which can also be efficiently used for sophisticated molecular techniques.

Analysis of the effect of various decalcification agents on the quantity and quality of nucleic acid (DNA and RNA) recovered from bone biopsies

2013 ◽  
Vol 17 (4) ◽  
pp. 322-326 ◽  
Author(s):  
Veena M. Singh ◽  
Ranelle C. Salunga ◽  
Vivian J. Huang ◽  
Yen Tran ◽  
Mark Erlander ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Dna And Rna ◽  
Bone Biopsies

scholarly journals Epitachophoresis is a novel versatile total nucleic acid extraction method

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Vladimira Datinska ◽  
Pantea Gheibi ◽  
Keynttisha Jefferson ◽  
Jaeyoung Yang ◽  
Sri Paladugu ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Nucleic Acid ◽  
Extraction Method ◽  
Acid Extraction ◽  
Total Nucleic Acid ◽  
Next Generation ◽  
Nucleic Acid Extraction ◽  
Dna And Rna ◽  
Generation Sequencing

AbstractEpitachophoresis is a novel next generation extraction system capable of isolating DNA and RNA simultaneously from clinically relevant samples. Here we build on the versatility of Epitachophoresis by extracting diverse nucleic acids ranging in lengths (20 nt–290 Kbp). The quality of extracted miRNA, mRNA and gDNA was assessed by downstream Next-Generation Sequencing.

High-resolution nucleic acid sequence mapping via in situ hybridization at the Electron Microscope level

1995 ◽  
Vol 53 ◽  
pp. 752-753
Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch
Keyword(s):  
Nucleic Acid ◽  
Interphase Nucleus ◽  
Genome Mapping ◽  
Precise Location ◽  
Electron Microscope Level ◽  
Rna Sequences ◽  
Fundamental Interest ◽  
Whole Cells ◽  
Dna And Rna

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.

scholarly journals Sulfur Modification in Natural RNA and Therapeutic Oligonucleotides

2021 ◽  
Author(s):  
Ya Ying Zheng ◽  
Ying Wu ◽  
Thomas Begley ◽  
Jia Sheng
Keyword(s):  
Nucleic Acid ◽  
Dna And Rna ◽  
Sulfur Modification ◽  
Oxygen Atoms

Sulfur modifications have been discovered on both DNA and RNA. Sulfur substitution of oxygen atoms at nucleobase or backbone locations in the nucleic acid framework led to a wide variety...

scholarly journals Deep learning-based transformation of H&E stained tissues into special stains

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kevin de Haan ◽  
Yijie Zhang ◽  
Jonathan E. Zuckerman ◽  
Tairan Liu ◽  
Anthony E. Sisk ◽  
...  
Keyword(s):  
Visual Inspection ◽  
Kidney Diseases ◽  
Periodic Acid ◽  
Needle Core Biopsy ◽  
Silver Stain ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Biopsy Tissue ◽  
Hematoxylin And Eosin

AbstractPathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson’s Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.

scholarly journals 2P123 Bacterial ribosomal RNA as a target for sequence-specific inhibition(05A. Nucleic acid: Structure & Property,Poster)

2013 ◽  
Vol 53 (supplement1-2) ◽  
pp. S179
Author(s):  
Joanna Tyylska ◽  
Sapna G. Thoduka ◽  
Zofia Dabrowska ◽  
Anna Gorska ◽  
Maciej Jasinski ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Ribosomal Rna ◽  
Specific Inhibition ◽  
Structure Property ◽  
Nucleic Acid Structure ◽  
Acid Structure
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