Flow cytometric characterization of microspore development in Brassica napus

1992 ◽  
Vol 70 (4) ◽  
pp. 802-809 ◽  
Author(s):  
K. Fuchs ◽  
K. P. Pauls
Keyword(s):  
Flow Cytometry ◽  
Brassica Napus ◽  
Light Scatter ◽  
Microspore Development ◽  
Size Category ◽  
Flow Cytometric ◽  
Key Words Flow Cytometry ◽  
Pollen Ontogeny ◽  
Bud Size

Flow cytometry was found to be sensitive to change in the physical characteristics of developing Brassica napus microspores. By measuring forward angle (10° – 19°) light scatter (FALS) and log 90° light scatter (L90LS) several stages of microspore development were characterized in small (1.5 mm to 2.5 mm), medium (2.5 mm to 3.5 mm), and large (3.5 mm to 4.5 mm) buds. Cell sorting was used to identify the types of cells that made up the subpopulations in two parameter plots of FALS versus L90LS including tetrad, translucent, trilobate, round, and oval microspores. Microspore development was found to be synchronous in the variety that was studied (‘Topas’) since only a few stages were found in each bud-size category. The study defined the normal process of pollen ontogeny in terms of the changes in light scatter that the cells underwent and demonstrated that flow cytometry is a useful method for analyzing this process. Key words: flow cytometry, Brassica napus, microspores, pollen, light scatter.

Diagnostic Utility of Flow Cytometry in Myelodysplastic Syndrome: A Retrospective Validation From a Single Centre.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4846-4846
Author(s):  
Pervin Topcuoglu ◽  
Klara Dalva ◽  
Sule Mine Bakanay ◽  
Sinem Civriz Bozdag ◽  
Onder Arslan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Myelodysplastic Syndrome ◽  
Antigen Expression ◽  
Pathological Examination ◽  
Light Scatter ◽  
Myeloid Lineage ◽  
Hematopoietic Stem ◽  
Altered Expression ◽  
Flow Cytometric

Abstract Abstract 4846 Myelodysplastic syndrome (MDS) is heterogeneous clonal hematopoietic stem cell disorder characterized by cytopenia(s) and dysplasia in one or more cell lineage. Though flow cytometry (FCM) is an important diagnostic tool in hematopoietic cell disorders, a prominent immunophenotyping feature in MDS may not be determined. In this study, we retrospectively evaluated flow cytometric features of bone marrow samples diagnosed as MDS with clinical and hematological findings. Patients-Method Between Feb 2004 and March 2009, flow cytometric parameters of 73 patients (M/F: 50/23) with MDS were re-analyzed. Median age was 59 years (17-89 ys). Our general principles are to evaluate quality of bone marrow samples, to determine proportion of the cells and features of their light scatter, and to give percentage of the blast. When detected a finding of dysplasia in the first analysis, the second step includes the determination of the maturation of the cells and the presence of the aberrant antigen expression. Results The samples were interpreted as MDS in % 76.7, MDS-RAEB-1 or RAEB-2 in %16.4, myeloproliferative disorder in %1.4 and non-diagnostic in %6.8 of the cases by flow cytometric examination. We detected variable degrees of hypogranulation in myeloid lineage in %82.2 of the samples by the light scatter features of the cells: 85% of severe and 15% of moderate or mild hypogranulation. The ratio of myeloid and lymphoid was changing from 0.3 to 17.5 (median 2). The decreasing of this ratio (<1) was observed in 19.4% of the samples. We detected altered expression of mature granulocyte. These included decreasing or lack of expression in CD15 45/73 (61.4%), CD13 38/70 (54.3%), CD16 53/67 (79.1%), CD11b 51/71 (71.8%), CD24 44/69 (65.2%), CD10 23/72 (31.9%) and MPO 14/72 (19.4%). Besides, bright expression of CD33 in 53.5% of the samples was observed. CD36 and CD56 in myeloid lineage were co-expressed in about 50 % of the samples. In 80.8 % of the samples dysplasia in erythroid compartment could be evaluated: Expression of CD71 according to glycophorin A (ratio <1) was decreased in 23.7 %. When we made similar analysis in the samples without RAEB-1 and -2 as pathological examination of bone marrow, 13.4 % of the samples could not be evaluated in favor of dysplasia. Of the samples with dysplasia hypogranulation, aberrant antigen expression of myeloid lineage and eryhtroid dysplasia were observed in 92.1%, 34.1% and 31.5%, respectively. In conclusion, FCM events may help to the differantial diagnosis of MDS especially when combining with clinical events. Improving of the analysis by focusing on the blast characteristics may be a standard approach to evaluate for low risk MDS. Disclosures No relevant conflicts of interest to declare.

A New Algorithm and Panel Construction for Pediatric Leukemia Immunophenotyping Using 10-Color Flow Cytometry

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4799-4799
Author(s):  
Bettina Keller ◽  
Markus P Radsak ◽  
Joerg Faber ◽  
Alexandra Russo
Keyword(s):  
Flow Cytometry ◽  
Childhood Leukemia ◽  
Conflicts Of Interest ◽  
High Reliability ◽  
Maturation Stage ◽  
Specific Information ◽  
Color Flow ◽  
Pediatric Leukemia ◽  
Flow Cytometric

Abstract Abstract 4799 Background: Rapid identification and quantification of abnormal cell populations in minimal specimen are crucial for diagnosis and longitudinal minimal residual disease (MRD) testing of childhood leukemia. So far, most standard immunophenotypic analyses are performed using antibody panels with up to five-colors and require high cell numbers. For infant and pediatric specimen, high-level multicolor analyses is highly desirable to gather sufficient data for initial diagnostic and follow up monitoring of pathologic populations. Objective: In this study, we aimed to establish a newly defined pediatric multicolor flow cytometric panel algorithm with high reliability yet minimal specimen requirement. Results: We defined a 10-color flow cytometric panel using the new violet laser dye “KromeOrange (KO)”. Applying CD45-KO/Side Scatter gating, combined with 2 additional backbone markers the panel is designed in two consecutive steps. In the first step, a single standardized 10-color-“screening tube” (FITC-HLA-DR, PE-CD15/CD56, ECD-CD5, PC5.5-CD33, PC7-CD13, APC-CD117, APC A700-CD34, APC A750-CD19, PB-CD3, KrO-CD45) is applied for initial orientation of specific lineage assignment. Based on results obtained with the screening tube, a specific multi-tube “classification panel” is used to complete detailed characterization of lineage specific malignancy and maturation stage. Suitable specimens include fresh blood, bone marrow and all body fluids. All samples are stained directly with monoclonal antibodies, followed by the lyses of erythrocytes and a short wash. Compared to standard five color panel previously used the application of greater numbers of informative antibodies in the screening tube and in the 2ndstep muti-tube classification panel is cost and time efficient and results in a more precise characterization of any single event. Conclusion: Our panel construction and algorithm definition for infant and pediatric leukemia immunophenotyping is one of the first 10-color flow cytometry panels described for this application. Advantages are the possibility to obtain highly specific information from minimal specimens with significantly improved laboratory efficiency. The overall performance is currently tested in a routine clinical setting. Disclosures: No relevant conflicts of interest to declare.

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

Abstract The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

Characterization of a pollen-specific gene family from Brassica napus which is activated during early microspore development

1990 ◽  
Vol 15 (4) ◽  
pp. 605-622 ◽  
Author(s):  
Diego Albani ◽  
Laurian S. Robert ◽  
Pauline A. Donaldson ◽  
Illimar Altosaar ◽  
Paul G. Arnison ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Family ◽  
Specific Gene ◽  
Microspore Development

scholarly journals Identification and Genetic Diversity Analysis of Edible and Medicinal Malva Species Using Flow Cytometry and ISSR Molecular Markers

Agronomy ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 650
Author(s):  
Iwona Jedrzejczyk ◽  
Monika Rewers
Keyword(s):  
Flow Cytometry ◽  
Molecular Markers ◽  
Genome Size ◽  
Inter Simple Sequence Repeat ◽  
Issr Markers ◽  
Size Estimation ◽  
Flow Cytometric ◽  
First Choice ◽  
Issr Molecular Markers

The Malva genus contains species that reveal therapeutic properties and are mostly important in medicine and the functional food industry. Its breeding, cultivation, and utilization are based on proper germplasm/plant identification, which is difficult using morphological features. For this reason, we applied flow cytometry and inter simple sequence repeat polymerase chain reaction (ISSR-PCR) for fast and accurate species identification. Genome size estimation by flow cytometry was proposed as the first-choice method for quick accession screening. Out of the 12 tested accessions, it was possible to identify six genotypes based on genome size estimation, whereas all species and varieties were identified using ISSR markers. Flow cytometric analyses revealed that Malva species possessed very small (1.45–2.77 pg/2C), small (2.81–3.80 pg/2C), and intermediate (11.06 pg/2C) genomes, but the majority of accessions possessed very small genomes. Additionally, this is the first report on genome size assessment for eight of the accessions. The relationships between the investigated accessions showed the presence of two clusters representing malvoid and lavateroid group of species. Flow cytometry and ISSR molecular markers can be effectively used in the identification and genetic characterization of Malva species.

scholarly journals The Value of Flow Cytometry Clonality in Large Granular Lymphocyte Leukemia

Cancers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 4513
Author(s):  
Valentina Giudice ◽  
Matteo D’Addona ◽  
Nunzia Montuori ◽  
Carmine Selleri
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Cell Receptor ◽  
Large Granular Lymphocyte ◽  
Effector Memory ◽  
Memory Cd8 T Cells ◽  
Flow Cytometric ◽  
Autoimmune Conditions

Large granular lymphocyte (LGL) leukemia is a lymphoproliferative disorder of mature T or NK cells frequently associated with autoimmune disorders and other hematological conditions, such as myelodysplastic syndromes. Immunophenotype of LGL cells is similar to that of effector memory CD8+ T cells with T-cell receptor (TCR) clonality defined by molecular and/or flow cytometric analysis. Vβ usage by flow cytometry can identify clonal TCR rearrangements at the protein level, and is fast, sensitive, and almost always available in every Hematology Center. Moreover, Vβ usage can be associated with immunophenotypic characterization of LGL clone in a multiparametric staining, and clonal kinetics can be easily monitored during treatment and follow-up. Finally, Vβ usage by flow cytometry might identify LGL clones silently underlying other hematological conditions, and routine characterization of Vβ skewing might identify recurrent TCR rearrangements that might trigger aberrant immune responses during hematological or autoimmune conditions.

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

scholarly journals Rugosity in Grimontia hollisae

2006 ◽  
Vol 73 (4) ◽  
pp. 1215-1224 ◽  
Author(s):  
S. K. Curtis ◽  
M. H. Kothary ◽  
R. J. Blodgett ◽  
R. B. Raybourne ◽  
G. C. Ziobro ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Wheat Germ ◽  
Cellular Differentiation ◽  
Lectin Binding ◽  
Single Cells ◽  
Light Scatter ◽  
Agar Surface ◽  
Galanthus Nivalis ◽  
Vibrio Hollisae

ABSTRACT Grimontia hollisae, formerly Vibrio hollisae, produces both smooth and rugose colonial variants. The rugose colony phenotype is characterized by wrinkled colonies producing copious amounts of exopolysaccharide. Cells from a rugose colony grown at 30�C form rugose colonies, while the same cells grown at 37�C form smooth colonies, which are characterized by a nonwrinkled, uncrannied appearance. Stress response studies revealed that after exposure to bleach for 30 min, rugose survivors outnumbered smooth survivors. Light scatter information obtained by flow cytometry indicated that rugose cells clumped into clusters of three or more cells (average, five cells) and formed two major clusters, while smooth cells formed only one cluster of single cells or doublets. Fluorescent lectin-binding flow cytometry studies revealed that the percentages of rugose cells that bound either wheat germ agglutinin (WGA) or Galanthus nivalis lectin (GNL) were greater than the percentages of smooth cells that bound the same lectins (WGA, 35% versus 3.5%; GNL, 67% versus 0.21%). These results indicate that the rugose exopolysaccharide consists partially of N-acetylglucosamine and mannose. Rugose colonies produced significantly more biofilm material than did smooth colonies, and rugose colonies grown at 30�C produced more biofilm material than rugose colonies grown at 37�C. Ultrastructurally, rugose colonies show regional cellular differentiation, with apical and lateral colonial regions containing cells embedded in a matrix stained by Alcian Blue. The cells touching the agar surface are packed tightly together in a palisade-like manner. The central region of the colony contains irregularly arranged, fluid-filled spaces and loosely packed chains or arrays of coccoid and vibrioid cells. Smooth colonies, in contrast, are flattened, composed of vibrioid cells, and lack distinct regional cellular differences. Results from suckling mouse studies showed that both orally fed rugose and smooth variants elicited significant, but similar, amounts of fluid accumulated in the stomach and intestines. These observations comprise the first report of expression and characterization of rugosity by G. hollisae and raise the possibility that expression of rugose exopolysaccharide in this organism is regulated at least in part by growth temperature.

scholarly journals Immunophenotypic Characterization of Human Bone Marrow Mast Cells. A Flow Cytometric Study of Normal and Pathological Bone Marrow Samples

1998 ◽  
Vol 16 (3) ◽  
pp. 151-159 ◽  
Author(s):  
Luis Escribano ◽  
Alberto Orfao ◽  
Jesús Villarrubia ◽  
Beatriz Díaz-Agustín ◽  
Carlos Cerveró ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Mast Cells ◽  
Antigen Expression ◽  
Hematologic Malignancies ◽  
Healthy Controls ◽  
Flow Cytometric ◽  
Hla Dr ◽  
Flow Cytometric Study

The goal of the present paper was to define the immunophenotype of bone marrow mast cells (BMMC) from healthy controls and patients with hematologic malignancies (HM) based on the use of multiple stainings with monoclonal antibodies analyzed by flow cytometry. Our results show that BMMC from both groups of individuals display a similar but heterogenous immunophenotype. The overall numbers of BMMC are higher in the HM group of individuals (p= 0.08). Three patterns of antigen expression were detected: (1) markers constantly positive in all cases analyzed (CD9, CD29, CD33, CD43, CD44, CD49d, CD49e, CD51, CD71, CD117, and FcεRI), (2) antigens that were constantly negative (CD1a, CD2, CD3, CD5, CD6, CD11a, CD14, CD15, CD16, CD19, CD20, CD21, CD23, CD25, CD30, CD34, CD38, CD41a, CD42b, CD65, CD66b, HLA-DR, and CD138), and (3) markers that were positive in a variable proportion of cases – CD11b (50%), CD11c (77%), CD13 (40%), CD18 (20%), CD22 (68%), CD35 (27%), CD40 (67%), CD54 (88%) and CD61 (40%). In addition, BMMC from all cases explored were CD45+, and this antigen was expressed at an intensity similar to that of mature granulocytes.In summary, our results show that BMMC from both healthy controls and HM patients display a relatively heterogenous immunophenotype. Interestingly, we have observed clear differences between the immunophenotype of BMMC and MC from other tissues. This could be due either to the heterogeneity of human MC according to their tissue localization or to the sensitivity of the method used for antigen detection.

Sign in / Sign up

Export Citation Format

Share Document