Identification and Cytogenetic Analysis of an Abnormal Pig Chromosome for Flow Cytometry and Sorting

1993 ◽  
Vol 48 (7-8) ◽  
pp. 645-653 ◽  
Author(s):  
Michael Hausmann ◽  
C. Paul Popescu ◽  
Jeannine Boscher ◽  
Dominique Kerboœf ◽  
Jürgen Dölle ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Marker Chromosome ◽  
Flow Analysis ◽  
Chromosome 1 ◽  
Cell Type ◽  
Metaphase Chromosomes ◽  
Flow Sorting ◽  
Dna Libraries ◽  
Sus Scrofa Domestica

Abstract For cytogenetics of pig (Sus scrofa domestica) and the influence of chromosome aberrations on pig production, high interest exists in flow sorted chromosomes for gene mapping, to estab­lish DNA-libraries, or to produce DNA-probes. Flow karyotyping and sorting as well as slit scan flow analysis of metaphase chromosomes of an abnormal cell type carrying a translocation marker chromosome 6/15 are described. Flow sorting of the largest chromosomes of these cells was performed. After sorting the chromosomes still had a well preserved morphology and were identified microscopically by G-banding. The quality of the band pattern of the sorted chromosomes was compatible to that of isolated chromosomes not subjected to flow cytometry. The sorted fraction showed an enrichment of chromosom e 6/15 and chromosome 1 which have quantitatively about the same integrated fluorescence intensity. Slit scan flow analysis was performed to discriminate these two chromosomes. Metacentric and submetacentric chromosom es were analyzed according to their bimodal slit scan profiles. Profiles of the largest chromosomes were distinguished by their different centromeric indices. Two groups were interpreted as the normal chromosome 1 and the translocation chromosom e 6/15.

scholarly journals Flow Cytometry and Flow Sorting of Metaphase Chromosomes from the Dasyurid Marsupial Dasyurus viverrinus

1985 ◽  
Vol 38 (4) ◽  
pp. 377 ◽  
Author(s):  
Brandon Wainwright ◽  
Rory Hope
Keyword(s):  
Flow Cytometry ◽  
Chromosomal Dna ◽  
Metaphase Chromosomes ◽  
Homogeneous Groups ◽  
Flow Sorting ◽  
Dna Contents ◽  
Conventional Methods

Metaphase chromosomes (2n = 14) from D. viverrinus were analysed by flow cytometry and flow sorted into six homogeneous groups. Relative chromosomal DNA contents and distribution frequencies of the groups corresponded closely with values for the karyotype obtained by conventional methods.

scholarly journals Reevaluating Patient Eligibility for Inotuzumab Ozogamicin Based on CD22 Expression: Is Dim Expression Sufficient?

2020 ◽  
Vol 28 (1) ◽  
pp. 252-259
Author(s):  
Warren Fingrut ◽  
Wendy Davis ◽  
Eric McGinnis ◽  
Karen Dallas ◽  
Khaled Ramadan ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Acute Lymphoblastic Leukemia ◽  
Survival Benefit ◽  
Lymphoblastic Leukemia ◽  
Surface Expression ◽  
Inotuzumab Ozogamicin ◽  
Cell Acute Lymphoblastic Leukemia

Salvage options for patients with relapsed B-cell acute lymphoblastic leukemia (B-ALL) include inotuzumab ozogamicin (InO), a recombinant, humanized anti-CD22 monoclonal antibody conjugated to the cytotoxic antibiotic calicheamicin. However, the benefit of InO in patients with dim CD22 expression remains unclear. We present a case of a patient with B-ALL who responded to InO despite only dim surface expression of CD22 by flow cytometry, achieving a survival benefit concordant with that reported in the literature and maintaining a good quality of life as a transfusion-independent outpatient. Our observation has broad relevance to clinicians who manage patients with B-ALL who are candidates for InO.

Cloning of DNA libraries from mouse Y chromosomes purified by flow cytometry

1986 ◽  
Vol 12 (3) ◽  
pp. 289-295 ◽  
Author(s):  
Bruno Baron ◽  
Philippe Métézeau ◽  
Didier Hatat ◽  
Christa Roberts ◽  
Michel E. Goldberg ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Y Chromosomes ◽  
Dna Libraries

scholarly journals Simultaneous localization and quantification of relative G and F actin content: optimization of fluorescence labeling methods.

1992 ◽  
Vol 40 (10) ◽  
pp. 1605-1612 ◽  
Author(s):  
G C Knowles ◽  
C A McCulloch
Keyword(s):  
Flow Cytometry ◽  
Confocal Microscopy ◽  
Actin Polymerization ◽  
Direct Method ◽  
Dnase I ◽  
Fluorescence Probes ◽  
Fluorescence Labeling ◽  
Cell Type ◽  
Fluorescence Spectrophotometry ◽  
Actin Rearrangement

Previous studies of fluorescence probes for labeling the monomeric actin pool have demonstrated lack of specificity. We have used quantitative analytical methods to assess the sensitivity and specificity of rhodamine DNAse I as a probe for monomeric (G) actin. The G-actin pool of attached or suspended fibroblasts was stabilized by ice-cold glycerol and MgCl2. Formaldehyde fixation was used to clamp the filamentous (F) actin pool. G- and F-actins were stained by rhodamine DNAse I and FITC-phalloidin, respectively. Confocal microscopy indicated that the G- and F-actins were spatially separate in substrate-attached cells. Flow cytometry and fluorescence spectrophotometry demonstrated low co-labeling of the separate actin pools, although measureable background binding of rhodamine DNAse I was detectable. Estimates of the extent of actin polymerization after trypsinization demonstrated reciprocal changes of monomeric and filamentous actins, consistent with the formation of a perinuclear array of F-actin. The labeling and quantitation methods were also sufficiently sensitive to detect cell type-dependent variations in actin content. Dual labeling of cells with rhodamine DNAse I and FITC-phalloidin may provide a simple and direct method to image and quantify actin rearrangement in individual cells.

scholarly journals A template method for decomposing flow cytometry histograms of human chromosomes.

1979 ◽  
Vol 27 (1) ◽  
pp. 305-310 ◽  
Author(s):  
D H Moore
Keyword(s):  
Flow Cytometry ◽  
Trisomy 21 ◽  
Chromosomal Abnormalities ◽  
Simulated Data ◽  
Normal Karyotype ◽  
Template Method ◽  
Normal Person ◽  
Human Chromosomes ◽  
Metaphase Chromosomes ◽  
Flow Measurements

A new method for decomposing flow cytometry histograms of isolated human metaphase chromosomes is described and tested. The method is based on fitting a template, composed of the means of all chromosomes of a normal karyotype to the flow histogram. The utility of the method is demonstrated by application to flow measurements of chromosomes from a normal person and comparing the results with those obtained by conventional cytophotometry. The power of the method for detecting gross chromosomal abnormalities, such as trisomy 21, as well as more subtle variations such as a single translocation, is determined for simulated data.

Development of a multiomics and functional drug sensitivity platform to investigate cell type specific drug effects in AML.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e19013-e19013
Author(s):  
Marianne T. Santaguida ◽  
Ryosuke Kita ◽  
Steven A. Schaffert ◽  
Erica K. Anderson ◽  
Kamran A Ali ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Drug Response ◽  
Drug Sensitivity ◽  
Ex Vivo ◽  
Cell Types ◽  
Specific Cell ◽  
Rna Seq ◽  
Cell Type ◽  
Drug Sensitivity Assay ◽  
Cell Type Specific

e19013 Background: Understanding the heterogeneity of AML is necessary for developing targeted drugs and diagnostics. A key measure of heterogeneity is the variance in response to treatments. Previously, we developed an ex vivo flow cytometry drug sensitivity assay (DSA) that predicted response to treatments in myelodysplastic syndrome. Unlike bulk cell viability measures of other drug sensitivity assays, our flow cytometry assay provides single cell resolution. The assay measures a drug’s effect on the viability or functional state of specific cell types. Here we present the development of this technology for AML, with additional measurements of DNA-Seq and RNA-Seq. Using the data from this assay, we aim to characterize the heterogeneity in AML drug sensitivity and the molecular mechanisms that drive it. Methods: As an initial feasibility analysis, we assayed 1 bone marrow and 3 peripheral blood AML patient samples. For the DSA, the samples were cultured with six AML standard of care (SOC) compounds across seven doses, in addition to two combinations. The cells were stained to detect multiple cell types including tumor blasts, and drug response was measured by flow cytometry. For the multi-omics, the cells were magnetically sorted to enrich for blasts and then assayed using a targeted 400 gene DNA-Seq panel and whole bulk transcriptome RNA-Seq. For comparison with BeatAML, Pearson correlations between gene expression and venetoclax sensitivity were investigated. Results: In our drug sensitivity assay, we measured dose response curves for the six SOC compounds, for each different cell type across each sample. The dose responses had cell type specific effects, including differences in drug response between CD11b+ blasts, CD11b- blasts, and other non-blast populations. Integrating with the DNA-Seq and RNA-Seq data, known associations between ex vivo drug response and gene expression were identified with additional cell type specificity. For example, BCL2A1 expression was negatively correlated with venetoclax sensitivity in CD11b- blasts but not in CD11b+ blasts. To further corroborate, among the top 1000 genes associated with venetoclax sensitivity in BeatAML, 93.7% had concordant directionality in effect. Conclusions: Here we describe the development of an integrated ex vivo drug sensitivity assay and multi-omics dataset. The data demonstrated that ex vivo responses to compounds differ between cell types, highlighting the importance of measuring drug response in specific cell types. In addition, we demonstrated that integrating these data will provide unique insights on molecular mechanisms that affect cell type specific drug response. As we continue to expand the number of patient samples evaluated with our multi-dimensional platform, this dataset will provide insights for novel drug target discovery, biomarker development, and, in the future, informing treatment decisions.

scholarly journals Morphological changes in colchicine--treated Pelargonium × hortorum L.H. Bailey greenhouse plants

2010 ◽  
Vol 37 (No. 1) ◽  
pp. 27-33 ◽  
Author(s):  
P. Jadrná ◽  
O. Plavcová ◽  
F. Kobza
Keyword(s):  
Flow Cytometry ◽  
Morphological Changes ◽  
Water Solutions ◽  
Breeding Material ◽  
Leaf Coloration

Polyploids were effectively pre-selected in colchicine-treated plants of the desirable brown-leaved cultivar Black Velvet Scarlet F1 of the species <I>Pelargonium × hortorum</I> L.H. Bailey to obtain the basic breeding material for creating new brown-leaved tetraploid cultivars. The green-leaved cultivar Gizela F1 was used for comparison of quantity and quality of response to colchicine treatments. Water solutions of colchicine in the range from 0.1% to 2.5% induced polyploidy in seedlings with treatments repeated each day for 2, 3, 5 or 7 days. Polyploid plants were pre-selected according to their morphological changes and stomata length and density and verified using flow cytometry. Some morphological changes (leaf coloration, flower shapes) in colchiploids differed between the genotypes, others were the same in both cultivars (loss of coloration in mixoploids, failure of blooming).

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