Morphometry to identify subtypes of leukocytes

Tozetti, P.B.; Lima, Ewelyne Miranda de; Nascimento, Andrews Marques do; Endringer, Denise Coutinho; Pinto, Fernanda E.; Andrade, Tadeu Uggere de; Mittag, Anja; Tárnok, Attila; Lenz, Dominik

doi:10.1016/j.hemonc.2013.11.005

Cited by 11 publications

(3 citation statements)

References 18 publications

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“…CP allows automated cellular identification and the analysis of hundreds of parameters to gain a plethora of information about intensity, morphology, and texture [13]. Furthermore, the software offers simultaneous analysis of different images (Carpenter et al, 2006) and a reproducible analysis [14, 15]. The CPA software has a machine learning-based classifier that can be used, e.g., to identify and count different cell types or cells in different phases of the cell cycle [13].…”

Section: Introductionmentioning

confidence: 99%

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

Macedo

Buzin

Araujo

et al. 2019

Journal of Immunology Research

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Background. Manual analysis of tissue sections, such as for pathological diagnosis, requires an analyst with substantial knowledge and experience. Reproducible image analysis of biological samples is steadily gaining scientific importance. The aim of the present study was to employ image analysis followed by machine learning to identify vascular endothelial growth factor (VEGF) in kidney tissue that had been subjected to hypoxia. Methods. Light microscopy images of renal tissue sections stained for VEGF were analyzed. Subsequently, machine learning classified the cells as VEGF+ and VEGF- cells. Results. VEGF was detected and cells were counted with high sensitivity and specificity. Conclusion. With great clinical, diagnostic, and research potential, automatic image analysis offers a new quantitative capability, thereby adding numerical information to a mostly qualitative diagnostic approach.

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Section: Introductionmentioning

confidence: 99%

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

Macedo

Buzin

Araujo

et al. 2019

Journal of Immunology Research

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“…[ 10 ] It also performs biological phenotypes measurements automatically and quantitatively from a number of images. [ 11 ] It is easy to be used by researchers who do not have great computer skills, good for implementation of methodology based on image analysis. [ 12 ] The program uses an algorithm to create a sequence of configurations to process the image according to the researcher's criteria.…”

Section: Introductionmentioning

confidence: 99%

In vitro cell viability by CellProfiler^®software as equivalent to MTT assay

et al. 2017

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Objective:This study evaluated in vitro cell viability by the colorimetric MTT stands for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay compared to image analysis by CellProfiler® software.Materials and Methods:Hepatoma (Hepa-1c1c7) and fibroblast (L929) cells were exposed to isolated substances, camptothecin, lycorine, tazettine, albomaculine, 3-epimacronine, trispheridine, galanthine and Padina gymnospora, Sargassum sp. methanolic extract, and Habranthus itaobinus Ravenna ethyl acetate in different concentrations. After MTT assay, cells were stained with Panotic dye kit. Cell images were obtained with an inverted microscope equipped with a digital camera. The images were analyzed by CellProfiler®.Results:No cytotoxicity at the highest concentration analyzed for 3-epimacronine, albomaculine, galanthine, trispheridine, P. gymnospora extract and Sargassum sp. extract where detected. Tazettine offered cytotoxicity only against the Hepa1c1c7 cell line. Lycorine, camptothecin, and H. itaobinus extract exhibited cytotoxic effects in both cell lines. The viability methods tested were correlated demonstrated by Bland–Atman test with normal distribution with mean difference between the two methods close to zero, bias value 3.0263. The error was within the limits of the confidence intervals and these values had a narrow difference. The correlation between the two methods was demonstrated by the linear regression plotted as R2.Conclusion:CellProfiler® image analysis presented similar results to the MTT assay in the identification of viable cells, and image analysis may assist part of biological analysis procedures. The presented methodology is inexpensive and reproducible.SUMMARYIn vitro cell viability assessment with MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay may be replaced by image analysis by CellProfiler®.The viability methods tested were correlated demonstrated by Bland-Atman test with normal distribution with mean difference between the two methods close to zero, bias value 3.0263. The correlation between the two methods was demonstrated by the linear regression plotted as R2.Abbreviations: HPLC: High pressure liquid chromatography MTT: (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide)

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“…Nevertheless, there are few studies available on Neotropical amphibian hematology (CABAGNA-ZENKLUSEN et al, 2005;CABAGNA-ZENKLUSEN et al, 2011;LAJMANOVÍCH et al, 2012), and scarce studies on anuran white blood cell morphology (ARIKAN; ÇIÇEK, 2010;. Few morphometric studies of red blood cells (BARNI et al, 1992;ATATÜR et al, 1999;WOJTASZEK;ADAMOWICZ, 2003;GRENAT et al, 2009) and white blood cells (FOXON 1964;NANO et al, 1991;TOZETTI et al, 2014) are available. To the authors' knowledge, this is the first study on white blood cell morphology in Neotropical anurans from South America.…”

Section: Introductionmentioning

confidence: 99%

Comparative leukocyte morphometric analysis between endemic Anurans from Brazil and the invasive species Lithobates catesbeianus

Silva

Ewbank

Strefezzi

et al. 2017

Braz. J. Vet. Res. Anim. Sci.

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Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry. Keywords: Anura. Morphometry. South America. White blood cells. ResumoAnfíbios são indicadores ambientais potencialmente confiáveis e eficientes. Estudos referentes à morfologia de leucócitos de anuros são limitados, com poucos estudos morfométricos disponíveis em literatura. O presente estudo empregou técnicas morfométricas para caracterizar leucócitos de anuros Neotropicais brasileiros selecionados e compará-los com a espécie exótica rã-touro (Lithobates catesbeianus), família Ranidae. Esfregaços sanguíneos de 28 espécimes pertencentes a seis gêneros diferentes (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus e Proceratophrys) foram comparados com amostras de esfregaços de L. catesbeianus. A média do diâmetro dos leucócitos foi calculada por um software de análise de imagens. One-way e teste de Bonferroni foram utilizados para avaliação estatística. Linfócitos, neutrófilos, eosinófilos e basófilos mostraram-se significativamente menores que os valores de referência reportados em outros gêneros de anfíbios, incluindo Lithobathes; por outro lado, a média do diâmetro dos monócitos não demonstrou variação significativa entre os gêneros. Esse é o primeiro estudo de avaliação morfométrica de leucócitos em espécies de anuros brasileiros. Nossos resultados sugerem que a separação geográfica possivelmente influencia a morfometria leucocitária. Palavras-chave: Anura. Morfometria. América do Sul. Leucócitos. | 160Braz.

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Morphometry to identify subtypes of leukocytes

Cited by 11 publications

References 18 publications

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

In vitro cell viability by CellProfiler^®software as equivalent to MTT assay

Comparative leukocyte morphometric analysis between endemic Anurans from Brazil and the invasive species Lithobates catesbeianus

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Morphometry to identify subtypes of leukocytes

Cited by 11 publications

References 18 publications

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

Automated and Reproducible Detection of Vascular Endothelial Growth Factor (VEGF) in Renal Tissue Sections

In vitro cell viability by CellProfiler®software as equivalent to MTT assay

Comparative leukocyte morphometric analysis between endemic Anurans from Brazil and the invasive species Lithobates catesbeianus

Contact Info

Product

Resources

About

In vitro cell viability by CellProfiler^®software as equivalent to MTT assay