Comparison of different staining methods for determination of viability on Mesocestoides vogae tetrathyridia

2018 ◽  
Vol 118 (2) ◽  
pp. 687-692 ◽  
Author(s):  
Julia Fabbri ◽  
María Celina Elissondo
Keyword(s):  
Staining Methods ◽  
Mesocestoides Vogae

scholarly journals Преимущества и возможности флуоресцентных методов для визуализации апоптоза и аутофагии в опухолевых клетках человека in vitro-=SUP=-*-=/SUP=-

2019 ◽  
Vol 126 (6) ◽  
pp. 771
Author(s):  
Н.А. Наволокин ◽  
Н.В. Полуконова ◽  
Д.А. Мудрак ◽  
А.М. Мыльников ◽  
М.А. Барышникова ◽  
...  
Keyword(s):  
Acridine Orange ◽  
Tumor Cells ◽  
Human Tumor ◽  
Double Staining ◽  
Chemotherapy Drugs ◽  
Autophagy Induction ◽  
Methods Of Evaluation ◽  
Staining Methods

The possibilities of using fluorescent research methods and their advantages for visualization and determination of the type of programmed cell death of human tumor cells under the action of flavonoids in experiments in vitro were investigated. The object of the study were the tumor cells of cervical cancer HeLa and A498 kidney carcinoma, the flavonoid containing extract of hedge hyssop (Gratiola officinalis L.) was used for testing in the experiments. The following fluorescent methods were used: the "living and dead" test with double staining - - iodide propidium and acridine orange, the method of double staining with annexin and iodide propidium. The confirmation of autophagy induction was performed using Muse cell analyzer with fluorescent reagents MuseAutophagy LC3-Antibody Based Kit. The application of fluorescent staining methods using double staining with acridine orange and iodide propidium in the "living and dead" test compared to phase microscopy allows to visualize the formation of apoptotic cells and autophagosomes in cells and, therefore, can serve as one of the methods of evaluation screening of the effectiveness of various chemotherapy drugs.

scholarly journals Determination of Coxiella burnetii in bovine foetuses using PCR and immunohistochemistry

2017 ◽  
Vol 61 (No. 8) ◽  
pp. 421-427 ◽  
Author(s):  
M. Ozkaraca ◽  
S. Ceribasi ◽  
AO Ceribasi ◽  
A. Kilic ◽  
S. Altun ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Coxiella Burnetii ◽  
Chain Reaction ◽  
Eastern Turkey ◽  
Staining Methods ◽  
Polymerase Chain ◽  
First Time

This study was aimed at detection of Coxiella burnetii in bovine foetuses using polymerase chain reaction (PCR) and immunohistochemistry (IHC) and at an estimation of its frequency in Eastern Turkey. Stamp, Giemsa, and Gimenez stains were used in addition to PCR and IHC to determine the presence of C. burnetii in samples from 70 bovine foetuses. While the staining methods did not detect the agent by direct visualisation of C. burnetii on smears, PCR and IHC identified its presence in two of the foetuses. The distribution of antigens in these two foetuses was, in decreasing order of concentration, in the spleen, the thymus, the lungs, the liver, and the kidneys. We conclude that C. burnetii diagnosis in bovine foetuses can be reliably performed using PCR and IHC. In addition, the frequency of 1.42% of C. burnetii positivity in bovine foetuses reported here was the first time that the presence of this agent was determined in Eastern Turkey.

scholarly journals Biofilm production by Listeria monocytogenes strains: detection with colorimetric analysis

2020 ◽  
Vol 30 (Supplement_5) ◽  
Author(s):  
P Centorame ◽  
L Iacone ◽  
R Salini ◽  
A Ciarulli ◽  
F Guidi ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Colorimetric Analysis ◽  
Solution Versus ◽  
Crystal Violet Solution ◽  
Biofilm Production ◽  
Staining Methods

Abstract Background In literature, there are no standardized laboratory methods to detect formed biomass by colorimetric analysis. The purpose of this study was to compare three staining methods and two different wavelengths for determination of biofilm formation of Listeria monocytogenes (Lm) strains. Methods Three strains of Lm isolated from different origin were tested using 96 well polistirene plates at 12 °C and 30 °C, after incubation the wells were subjected to washing, detaching and staining with crystal violet (CV) at 0.2% and 2% (Panreac EU) in 95% ethanol and with Gram's crystal violet solution (Merck KGaA, Germany). The absorbance at 492nm and 540nm wavelengths was read using a spectrophotometer (SIRIO S, Seac, Firenze, Italia). Results The strains incubated at 12 °C displayed production of biofilm when stained with CV 2% and with Gram's crystal violet solution, both at 492 and 540 nm (with better evidence at 540 nm). If CV 0.2% was used to stain and reading at both optical densities there was evidence of weak or no biofilm production. At 30 °C, the biofilm production was displayed at both temperature and with all the stains. For all the strains and for all the conditions tested, the absorbance was greater but not proportional using the Gram's crystal violet solution, versus the CV 0,2% and CV 2%, and absorbance was higher at 540nm versus at 492nm. Conclusions Results confirmed the lack of reproducibility of each of the method used to detect and quantify the biomass produced during a biofilm formation test in vitro and the absence of ratio between the different results obtained using different CV concentration and wavelengths for reading. Key messages Biofilm production at 12 °C could not be adequately detected staining the wells with CV 0,2%. Absorbance could be influenced by the solvent in the stain used (ethanol, methanol or phenol or mixtures). To obtain data for assessment of biomass formation, being the method characterized by poor reproducibility, the laboratory should use at least the same stain and wavelength.

scholarly journals Pewarnaan Kromosom dan Pemanfaatannya dalam Penentuan Tingkat Ploidi Eksplan Hasil Kultur Anter Anthurium

2016 ◽  
Vol 21 (2) ◽  
pp. 113 ◽  
Author(s):  
Budi Winarto
Keyword(s):  
Anther Culture ◽  
Ploidy Level ◽  
Staining Method ◽  
Root Tip ◽  
Root Tips ◽  
Staining Methods ◽  
Ornamental Crops ◽  
High Benefit ◽  
Chromosome Staining

Metode pewarnaan Kromosom yang optimal merupakan prasarat penting dalam penentuan level ploidi tanaman hasil kultur anter, termasuk variasi eksplan hasil kultur anter Anthurium. Aplikasi dan modifikasi metode pewarnaan kromosom pada berbagai eksplan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Februari sampai dengan Agustus 2009 untuk mengetahui keragaman dan tingkat ploidi regeneran hasil kultur anter Anthurium. Penelitian bertujuan mendapatkan metode pewarnaan kromosom dan modifikasinya, jenis eksplan dan akar yang sesuai untuk mempelajari tingkat ploidi regeneran hasil kultur anter Anthurium. Bahan yang digunakan ialah kalus, pucuk tunas, dan ujung akar udara. Penelitian terdiri atas tiga kegiatan, yaitu (1) modifikasi metode pewarnaan kromosom, (2) seleksi eksplan yang sesuai untuk pewarnaan kromosom, dan (3) optimasi metode pewarnaan kromosom terseleksi. Hasil penelitian menunjukkan bahwa ujung akar dan akar yang ditumbuhkan pada medium yang mengandung 1% arang aktif merupakan jenis eksplan dan akar yang sesuai untuk mendapatkan hasil pewarnaan kromosom yang baik. Modifikasi metode pewarnaan kromosom dengan pemanasan ujung akar pada 1N HCl : asam asetat glasial 45% (3:1, v/v) selama 10 menit pada suhu 60oC dan perlakuan aseto-orcein selama 15 menit merupakan metode pewarnaan kromosom yang lebih baik dalam menghasilkan obyek kromosom yang mudah dihitung. Penerapan metode pewarnaan kromosom pada kultur anter Anthurium dapat memisahkan tingkat ploidi regeneran. Pada penelitian ini rasio ploidi regeneran kultur anter ialah 33,5% haploid, 62,7% diploid, dan 5,7% triploid. Metode pewarnaan kromosom yang berhasil dikembangkan dalam penelitian ini sangat bermanfaat dalam pengembangan teknologi haploid pada jenis Araceae yang lain.<br /><br />Optimal chromosome staining method is important pre-requisite in determination of plant ploidy level derived from anther culture, involving varied explants regenerated from Anthurium anther culture. Application and modification of chromosome staining methods on different explants were conducted at the Tissue Culture Laboratory of  Indonesian Ornamental Crops Research Institute from February to August 2009 for determination of the ploidy level of regenerants derived from anther culture of Anthurium. The aim of this research was to determine the chromosome staining method and its modifications, type of explant and root suitable to study the ploidy level of explants derived from anther culture of Anthurium. Callus, shoot tips, and root tips were utilized in the experiment. The research was consisted of three experiments, i.e. (1) modification of chromosome staining methods (2) selection of explants suitable for chromosome staining, and (3) improvement of the selected chromosome staining method. Results of the study indicated that root tips and roots cultured on medium containing 1% active carchoal were the most appropriate explants and the root type in obtaining better chromosome staining results. The modification method with root tip boiled in 1N HCl : 45% of acetic acid glacial (3:1, v/v) for 10 minutes in 60ºC and aceto-orcein treatment for 15 minutes gave appropriate chromosome staining results exhibited clearer chromosome pictures and was easy to be counted. The  application of chromosome staining on anther culture of Anthurium was able to distinguish the ploidy level of regenerants. Ploidy ratio of regenerants derived from anther culture was 33.5% of haploid, 62.7% of diploid, and 5.7% of triploid. Chromosome staining method resulted from the study give high benefit in developing haploid technologies on other Araceae plants.<br /><br />

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