scholarly journals CHROMOSOME COUNT ON YOUNG ANTHER OF BANANA MALE BUD USING EZYMATIC MACERATION AND DAPI STAINING IN SLIDE PREPARATION

2020 ◽  
Vol 19 (2) ◽  
Author(s):  
Fajarudin Ahmad ◽  
Yuyu Suryasari Poerba
Keyword(s):  
Genetic Study ◽  
Cell Spreading ◽  
Enzymatic Digestion ◽  
Chromosome Count ◽  
Root Tip ◽  
Chromosome Counting ◽  
Fluorescent Staining ◽  
Dapi Staining ◽  
Slide Preparation ◽  
Staining Methods

Chromosome counting is the basis in describing the chromosomes number of organism that might useful for genetic study and classification. In banana studies, the root tip with a combination of non-fluorescent staining methods such as carmine or orcein and squash is the most common material for chromosome counting. In this study, we presented the usefulness of young anther of banana male bud with enzymatic maceration method for cell spreading and 4,6-diamino-2-phenyl-indole (DAPI) for staining agent to get a satisfying chromosomes image at metaphase for mitotic study of diploid and tetraploid bananas. The principle of this study is fixation using ethanol:acetic acid (3:1), enzymatic digestion, maceration and staining using DAPI. Our result showed that this method can provide well spread cells with intensely contrast of chromosomes images that satisfying for chromosome counting. 

scholarly journals AN IMPROVED FIXING AND STAINING PROCEDURE FOR PLOIDY DETERMINATIONS FROM ROOT TIP SQUASHES

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 646e-646
Author(s):  
Henry R. Owen ◽  
A. Raymond Miller
Keyword(s):  
Hydrochloric Acid ◽  
Root Tip ◽  
Staining Procedure ◽  
Chromosome Counting ◽  
Fragaria Ananassa ◽  
Polyploid Species ◽  
Treatment Combination ◽  
Root Tips ◽  
Carbol Fuchsin ◽  
Staining Methods

A comparison of pretreatment, fixing, and staining methods for root tips of Fragaria × ananassa (2n=8x=56), a polyploid species with small chromosomes, was made to facilitate chromosome counting. Three pretreatments (8-hydroxyquinoline, α-bromonaphthalene, and p-dichlorobenzene), three fixatives (Farmer's, Carnoy's, and Newcomer's), and five stains (acetocarmine, lacto-propionic orcein, leucobasic fuchsin, altered carbol fuchsin, and alcoholic hydrochloric-acid carmine) were examined in a factorial design to determine which treatment combination produced the best chromosome preparation. Field propagated runners were grown in sand under greenhouse conditions with supplemental lighting to produce root tips for late morning collection. The treatment combinations of α-bromonaphthalene or 8-hydroxyquinoline, Farmer's fixative, and altered carbol fuchsin, or the combination of α-bromonaphthalene, Farmer's fixative, and alcoholic hydrochloric-acid carmine produced the most intensely-stained and well-defined preparations.

scholarly journals Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Thaís Cristina Ribeiro Silva ◽  
Isabella Santiago Abreu ◽  
Carlos Roberto Carvalho
Keyword(s):  
Flow Cytometry ◽  
Root Meristem ◽  
Chromosome Analysis ◽  
Enzymatic Digestion ◽  
Root Tip ◽  
Alternative Source ◽  
Single Root ◽  
Nuclei Isolation ◽  
Flow Cytometric ◽  
Mechanical Isolation

Root meristems have increasingly been target of cell cycle studies by flow cytometric DNA content quantification. Moreover, roots can be an alternative source of nuclear suspension when leaves become unfeasible and for chromosome analysis and sorting. In the present paper, a protocol for intact nuclei isolation from a single root meristem was developed. This proceeding was based on excision of the meristematic region using a prototypical slide, followed by short enzymatic digestion and mechanical isolation of nuclei during homogenization with a hand mixer. Such parameters were optimized for reaching better results. Satisfactory nuclei amounts were extracted and analyzed by flow cytometry, producing histograms with reduced background noise and CVs between 3.2 and 4.1%. This improved and reproducible technique was shown to be rapid, inexpensive, and simple for nuclear extraction from a single root tip, and can be adapted for other plants and purposes.

Detection and Analysis of Intracellular Polymers Stored by Polyphosphate-Accumulating Microorganisms

2013 ◽  
Vol 777 ◽  
pp. 314-317
Author(s):  
Yan Hui Ge ◽  
Lin Zhao ◽  
Yan Zhou
Keyword(s):  
Quantitative Analysis ◽  
Qualitative And Quantitative Analysis ◽  
Dapi Staining ◽  
Qualitative And Quantitative ◽  
P Content ◽  
Staining Methods ◽  
Polyphosphate Accumulating Organisms

Both qualitative and quantitative analysis of intracellular polymers in polyphosphate accumulating organisms (PAOs) was discussed in this paper. Results were indicated that black-blue particles of poly-β-hydroxyalkanoate (PHBs) were observed clearly by Sudan black staining. DAPI staining and the Neisser new staining for polyphosphate (poly-P) were more proper than other staining methods. PHB content reached 198.3 mg/g MLSS detected by GC. After digested by HClO4, the poly-P content of sludge was determined to be 138.4 mg/g MLSS.

Conservation of feline semen: Part II: Cold-induced damages on spermatozoal fertilizing ability

2003 ◽  
Vol 5 (5) ◽  
pp. 257-263 ◽  
Author(s):  
G.C Luvoni ◽  
E Kalchschmidt ◽  
G Marinoni
Keyword(s):  
Special Reference ◽  
Functional Status ◽  
Limiting Factor ◽  
Fluorescent Staining ◽  
Epididymal Sperm ◽  
Freeze Thaw ◽  
Functional Integrity ◽  
Sperm Characteristics ◽  
Fertilizing Ability ◽  
Staining Methods

During the conservation of feline semen, the freeze–thaw procedure in particular is responsible for inducing severe spermatozoal damage, which diminishes fertilizing ability. Therefore, cold-induced damage represents a limiting factor for the conservation of semen, particularly semen from felids, which are often affected by teratospermia. In this article, feline sperm characteristics are reported, with special reference to motility and morphology, which are more likely to be affected by conservation protocols; and moreover, the causes of cold-induced damages are described. Attention has been focused on methods to evaluate functional integrity of spermatozoa, and those applied to cat semen are reviewed. Among these, a rather recently developed technique involves fluorescent staining methods, and in particular chlortetracycline. The chlortetracycline assay applied to cryopreserved cat epididymal sperm shows that it is suitable to evaluate the functional status of cat sperm.

scholarly journals Chromosome count on banana root tip squashes

Fruits ◽  
2008 ◽  
Vol 63 (3) ◽  
pp. 179-181 ◽  
Author(s):  
Frédéric Bakry ◽  
Kenneth Shepherd
Keyword(s):  
Chromosome Count ◽  
Root Tip

scholarly journals Diagnosis of Primary Cutaneous Amyloidosis by Rapid 4,6-Diamidino-2-Phenylindole Staining

Dermatology ◽  
2021 ◽  
pp. 1-9
Author(s):  
Junchen Chen ◽  
Huan Yang ◽  
Zhijun Xu ◽  
Ping Lu ◽  
Liyan Yuan ◽  
...  
Keyword(s):  
Congo Red ◽  
Positive Staining ◽  
Dapi Staining ◽  
Prurigo Nodularis ◽  
Tissue Sections ◽  
Double Stranded Dna ◽  
Experience Levels ◽  
Standard Procedures ◽  
Staining Methods ◽  
Congo Red Staining

<b><i>Background:</i></b> Quick and accurate diagnosis of primary cutaneous amyloidosis (PCA) may be difficult because its symptoms are often subtle and nonspecific. <b><i>Objective:</i></b> We sought to review the literature on the roles of various staining methods in the diagnosis of amyloidosis and demonstrate added benefits of using rapid 4,6-diamidino-2-phenylindole (DAPI) staining in the diagnosis of PCA. <b><i>Methods:</i></b> Three groups of cases, namely, PCA, neurodermatitis, and prurigo nodularis, were retrieved from a computerized pathology database for study, and their paraffin-embedded tissue blocks were cut following standard procedures. The tissue sections were stained with three stains: hematoxylin-eosin (HE), Congo red, and DAPI stains, and examined under the microscope to compare the staining patterns of these three methods. We also performed amyloid keratin and apolipoprotein E (APOE) staining on the sections of PCA in order to further support our conclusion. The PCA sections were read by junior and senior dermatopathologists for comparison. <b><i>Results:</i></b> The sensitivity of DAPI staining for PCA was significantly higher than that of Congo red staining and HE staining (<i>p</i> &#x3c; 0.001). This statement holds true whether the experiment was grouped in one sample or was divided into groups of junior and senior dermatopathologists (<i>p</i> &#x3c; 0.001). The DAPI-positive staining areas, except for the nuclei, were consistent with the amyloid deposition areas. In this study, DAPI staining had a sensitivity of 98.6% and a specificity of 100%. <b><i>Conclusion:</i></b> DAPI staining could serve as a useful technique to establish the diagnosis of PCA, and its high efficacy in diagnosing PCA makes it less dependent on the experience levels of the evaluators. Additionally, the binding of DAPI to the A-T-rich sequence of double-stranded DNA suggests that amyloid may contain DNA or a similarly structured nucleic acid.

scholarly journals The genetic effect of N-methyl-N-nitro-N-nitrosoguanidine on callus induced from mature embryos of beans Phaseolus vulgaris

2014 ◽  
Vol 8 (4) ◽  
pp. 79-84
Author(s):  
Sattar A. Shlahi ◽  
Zahra N. Al- Hattab
Keyword(s):  
Indole Acetic Acid ◽  
Casein Hydrolysate ◽  
Genetic Effect ◽  
Chromosome Count ◽  
Root Tips ◽  
Benzyl Adenine ◽  
Mature Embryos ◽  
Normal Number ◽  
Wide Range ◽  
Staining Methods

This research was conducted to study the effect of the chemical mutagen N-methyl-N-nitro-N-nitrosoguanidine (NTG)on the chromosomes of callus induced from mature bean embryos, Harvester cultivar. Seeds were treated with 0.2 or o.4 mM of the mutagen that mixed with different percentages of ethanol for 24 hrs. Calli were induced on MS medium in the presence of 0.5 mg/L of Benzyl adenine (BA), 1 mg/L Indole acetic acid (IAA) and 100 mg/L from each of Casein hydrolysate, Glycine, Asparagine, Tyrosine, and Myo-Inositol. Samples were pretreated with 1, 2- benzene dichloride, Para –dichlorobenzene, or Colchicine. Two different staining methods were used to stain the chromosomes from root tips and calli.The results showed that Para –dichlorobenzene is the best pretreatment for both root tips and callicells. However, the stain acetoorcein was the best for the root tips while Feulgen stain was the best for calli cells. Chromosome count showed that there were 22 chromosomes in all the cells of bean root tips (control). While a wide range of chromosome numbers were obtained from calli cells with or without mutagen treatment. Ninety six percent of the non treatedcalli gave the normal number of chromosomes while only 60% of calli treated with (0.4 mM+4% ethanol) gave the normal number of chromosomes. Calli cells from all the treatments showed chromosome multiplication except in the presence of ethanol.

scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTRY; II. THE DEMONSTRATION OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS USING FLUORESCEIN-LABELLED DEACETYLATED CHITIN

1962 ◽  
Vol 10 (1) ◽  
pp. 14-18 ◽  
Author(s):  
SIDNEY P. KENT ◽  
E. EDWARD EVANS
Keyword(s):  
Connective Tissue ◽  
Acridine Orange ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Fluorescent Staining ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Acidic Polysaccharides ◽  
Staining Methods ◽  
Formalin Fixed

A method for demonstrating acidic polysaccharides in formalin fixed parafin embedded tissue sections using fluorescein labelled deacetylated chitin is described. Epithelial and connective tissue acidic polysaccharides have been studied in a number of organs. The distribution of acidic polysaccharides as seen with fluorescein-labelled deacetylated chitin is similar to results obtained with alcian blue-periodic acid Schiff and fluorescein-labelled Aspergillus polysaccharide. Previously reported fluorescent staining methods for acidic polysaccharides such as iron-hematoxylin-acridine orange and atabrine as well as fluorescein-labelled deacetylated chitin may prove to be useful adjuncts to the light microscopic methods of demonstrating mucins. Other possible uses of deacetylated chitin, a colorless macro-cation, in histochemistry and cytochemistry are noted.

scholarly journals COMPARISON OF ZIEHL-NEELSEN, KINYOUN’S AND FLUORESCENT STAINING FOR DETECTION OF MYCOBACTERIUM TUBERCULOSIS IN SPUTUM SAMPLES BEFORE AND AFTER PETROFF’S CONCENTRATION TECHNIQUE

2018 ◽  
Vol 11 (4) ◽  
pp. 110
Author(s):  
Neelu Sree P ◽  
Terin J ◽  
Himadri Dutta ◽  
Kalyani M
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sputum Sample ◽  
Diagnostic Value ◽  
Fluorescent Staining ◽  
Concentration Technique ◽  
Before And After ◽  
Auramine O ◽  
Staining Methods ◽  
Staining Techniques ◽  
Microscopy Examination

 Objectives: The objectives of the study were to find out the presence of Mycobacterium tuberculosis in sputum samples using Ziehl-Neelsen (ZN), Kinyoun’s, and (auramine) fluorescent staining and to compare the three staining techniques with and without Petroff’s concentration and to find out the most preferable staining of M. tuberculosis. Methods: Sputum sample was collected and concentrated by 4% NaOH (Petroff’s concentration). Microscopic examination of the sample was done before concentration and after concentration by ZN staining, Kinyoun’s staining, and fluorescent staining (Auramine). Grading of acid-fast bacilli (AFB) by three staining was done before and after concentration according to RNTCP guidelines.Results: Total of 452 sputum samples were collected and subjected to microscopy examination by ZN, Kinyoun’s, and fluorescent staining methods to compare the presence or absence of AFB with or without concentration. Among the 452 sputum samples, total of 67 (15.6%) sputum samples were positive for the presence of AFB. Majority 40.3% (n=21) of TB positive patients were observed in the age group 51–60 years. The results of Auramine-O staining showed positive diagnoses in 15.9% of the samples; sensitivity was 100% and specificity 95.6%.Conclusion: The use of fluorescent staining significantly increases the diagnostic value of the smear, particularly where there are low-density bacilli which may escape detection on ZN stained smears.

Highly effective cell synchronization in plant roots by hydroxyurea and amiprophos-methyl or colchicine

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 387-390 ◽  
Author(s):  
W. H. Pan ◽  
A. Houben ◽  
R. Schlegel
Keyword(s):  
Dna Hybridization ◽  
Enzymatic Digestion ◽  
Root Tip ◽  
Chromosomal Dna ◽  
Cell Synchronization ◽  
Ice Water ◽  
Minimum Distortion ◽  
Barley Cell ◽  
Chromosome Spreading

Effective somatic cell synchronization in root-tip meristems and improved chromosome spreading were achieved in white campion, wheat, rye, and barley by application of hydroxyurea and amiprophos-methyl or colchicine, combined with a pretreatment of ice water and modified fixative, as well as enzymatic digestion of the meristems. The protocol provides metaphase indices of approximately 50%. The chromosomes and chromosomal DNA were with minimum distortion, providing useful material for chromosome banding studies, in situ DNA–DNA hybridization, microdissection, and microcloning.Key words: Melandrium album, rye, wheat, barley, cell cycle, root meristem, synchronization, metaphase index, chromosome preparation.

Sign in / Sign up

Export Citation Format

Share Document