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When monocytes and platelets compete: The effect of platelet count on the flow cytometric measurement of monocyte CD36Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Flow cytometric measurement of monocyte surface CD36 is relevant to several conditions including diabetes, cardiovascular disease, lipid disorders, platelet isoimmunization, and susceptibility to P falciparum malaria. CD36 is also strongly expressed on platelets where it is also known as platelet glycoprotein IV.Whole blood samples, containing identical monocyte concentrations, were adjusted to contain platelets ranging from 20,000/uL to 600,000/uL, were stained with fluorescent-labeled anti-CD36, and analyzed by flow cytometry.CD36 median fluorescent intensity (MFI) observed on monocytes decreased as the platelet concentr...
Source: Cytometry Part B: Clinical Cytometry - November 10, 2009 Category: Molecular Biology Authors: W.H. Dzik, C.M. Cserti-Gazdewich, I. Ssewanyana, M. DeLelys, F.I. Preffer Source Type: journals

Flow cytometric measurements of TB-specific T cells comparing with QuantiFERON-TB goldEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Interferon-[gamma] (IFN-[gamma]) release assays and the detection of IFN-[gamma] synthesis in the cytoplasm of activated CD4+ T cells by flow cytometry have recently been used for tuberculosis (TB) diagnosis. The aim of this study was to compare the performance of IFN-[gamma] assay between ELISA (QuantiFERON-TB Gold, QFT) and intracellular cytokine flow cytometry (ICCFC), and to investigate the significance of an optimal gating strategy in ICCFC.The CD4+ T cell response to TB antigens was measured using the intracellular cytokine staining technique and four color FC (CD3, CD4, IFN-[gamma], and tumor necrosis factor-[alpha]...
Source: Cytometry Part B: Clinical Cytometry - November 9, 2009 Category: Molecular Biology Authors: Dong Il Won, Jung Ran Park Source Type: journals

Evaluation of a dry format reagent alternative for CD4 T-cell enumeration for the FACSCount system: A report on a Moroccan-Canadian studyEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Efforts to improve alternative CD4 T-cell counting methods are critical to accelerate the implementation of HIV antiretroviral therapy in resources limited regions. Substituting liquid format reagents to eliminate cold-chain transportation and refrigerated storage with dry format reagents contributes to higher efficiency supply management solution especially for laboratories at remote locations. ReaMetrix has developed dry format reagent kits compatible with the FACSCount system, a dedicated flow cytometer for T-cell subset enumeration widely used in resource limited settings. A dual site collaborative study was designed t...
Source: Cytometry Part B: Clinical Cytometry - October 21, 2009 Category: Molecular Biology Authors: Michèle Bergeron, Tao Ding, Elmir Elharti, Hicham Oumzil, Nadia Soucy, Hicham Harmouche, Saad Chaouch, Rajae EL Aouad, Christian Chabot, Francis Mandy Source Type: journals

CD146+ T lymphocytes are increased in both the peripheral circulation and in the synovial effusions of patients with various musculoskeletal diseases and display pro-inflammatory gene profilesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Twenty-eight synovial effusions (SE) were obtained from 24 patients, paired samples of peripheral blood (PB) from 10 of these patients, and PB from 36 healthy individuals for analysis of CD146 on T-lymphocytes by flow cytometry. CD146+ or CD146- T-lymphocytes were sorted from three SE to study gene expression profiles and selected genes revalidated using QPCR assays. We found more CD3+CD146+ and CD4+CD146+ T-lymphocytes in PB from patients compared with PB of healthy individuals (4.71% ± 2.48% vs. 2.53% ± 1.08%, P = 0.028) and (6.29% ± 2.74% vs. 2.41% ± 0.96%, P = 0.0017), respectively, whereas CD8+CD146+ T-lymphocytes...
Source: Cytometry Part B: Clinical Cytometry - October 15, 2009 Category: Molecular Biology Authors: Pradeep Kumar Dagur, Gulnaz Tatlici, Mark Gourley, Leigh Samsel, Nalini Raghavachari, Poching Liu, Delong Liu, J. Philip McCoy Jr. Source Type: journals

Cytometric assessment of cytostatic and cytotoxic effects of topical glaucoma medications on human epithelial corneal line cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We examined the effect of glaucoma topical medications (bimatoprost, travoprost, latanoprost, timolol, betaxolol, dorzolamide, brinzolamide, brimonidine) on growth of cells of three human epithelial corneal lines.The cells were cultured in 8-chamber slides, treated with different concentrations of the medications, and fixed at 24, 48, and 72 h. Cell number on slides to estimate viability and growth curves, frequency of apoptosis (FLICA and caspase-3 activation probes), and proliferation (BrdU incorporation assay) were measured by laser scanning cytometry (LSC).Depending on concentration all examined medications induced cel...
Source: Cytometry Part B: Clinical Cytometry - October 14, 2009 Category: Molecular Biology Authors: Dorota Pozarowska, Piotr Pozarowski, Zbigniew Darzynkiewicz Source Type: journals

Variability of CD3 membrane expression and T cell activation capacityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In conclusion, T cells have highly heterogeneous CD3 expression, possibly predetermined and with clear functional significance. © 2009 Clinical Cytometry Society (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - October 14, 2009 Category: Molecular Biology Authors: Fatima-Zahra El Hentati, Frederic Gruy, Cristina Iobagiu, Claude Lambert Source Type: journals

Comparison of interlaboratory variation in absolute T-cell counts by single-platform and optimized dual-platform methodsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Previous studies have reported that the adoption of a single-platform flow cytometry cell counting method resulted in lower interlaboratory variation in absolute T cell counts as compared to predicate dual-platform flow cytometry methods which incorporate independent automated lymphocyte counts (Schnizlein-Bick et al., Clin Diagn Lab Immunol 2000;7:336-343; Reimann et al., Clin Diagn Lab Immunol 2000;7:344-351). In the present study, we asked whether use of a single-platform method could reduce variation in absolute cell counts across the laboratories in the Multicenter AIDS Cohort Study (MACS) (n = 4), as suggested by the...
Source: Cytometry Part B: Clinical Cytometry - October 7, 2009 Category: Molecular Biology Authors: Lance E. Hultin, Marianne Chow, Beth D. Jamieson, Maurice R. G. O'Gorman, Frederick A. Menendez, Luann Borowski, Thomas N. Denny, Joseph B. Margolick Source Type: journals

Utility of lyophilized PMA and ionomycin to stimulate lymphocytes in whole blood for immunological assaysEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The need to implement robust biomarkers in clinical trials has never been greater, and such efforts can be easily compromised by reagent instability or simple human error during assay set-up. Many biotechnology and pharmaceutical companies are introducing efforts to conduct biomarker studies under more rigorous settings, and the use of plates or tubes pre-loaded with stimulation or staining reagents could be of value for studies that involve flow cytometry.Five reagents lyophilized from ethanol or CHAPS buffer stock solution of phorbol 12-myristate 13-acetate (PMA) and ionomycin were benchmarked against standard DMSO liqui...
Source: Cytometry Part B: Clinical Cytometry - September 23, 2009 Category: Molecular Biology Authors: Shelley Sims Belouski, Julie Wilkinson, John Thomas, Keith Kelley, Shen-Wu Wang, Sid Suggs, John Ferbas Source Type: journals

Use of flow cytometry as a quality control device for liquid-based cervical cytology specimensEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The objective of the study is the evaluation of flow cytometry as a rapid tool for quality control of the liquid specimen adequacy for the purpose of precise cytological diagnosis in detecting cervical abnormalities.A total of 1,854 women undergoing routine cervical cytology joined this cohort study. A monolayer smear was prepared and 1 ml aliquot was removed and prepared for flow cytometric analysis. Cells were fixed and run on a Partec CyFlow SL, with front scatter (FSS) and side scatter (SSC) set on logarithmic scale. The forward scatter versus side scatter dot-plot was used for the distinction of ectocervical, endocerv...
Source: Cytometry Part B: Clinical Cytometry - September 10, 2009 Category: Molecular Biology Authors: Christine Kottaridi, John Georgoulakis, Dimitrios Kassanos, Asimakis Pappas, Aris Spathis, Niki Margari, Dionissios Aninos, Petros Karakitsos Source Type: journals

Frequency of circulating iNKT cells among Iranian healthy adultsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Invariant natural killer T cells (iNKT) are a small subset of T lymphocytes which are involved in a wide variety of immune responses. Human studies have reported a wide variation in the number of circulating iNKT cell among healthy donors as well as in some immunologic disorders while there is no or little data about the factors which affect the pool of circulating iNKT cells in human.Given the importance of iNKT cells in immunologic responses and variation in the number and phenotype of iNKT cells, the aim of this study was to evaluate the frequency of circulating iNKT cells in a cohort of healthy Iranian adults.The numbe...
Source: Cytometry Part B: Clinical Cytometry - August 27, 2009 Category: Molecular Biology Authors: Mohammad Fereidouni, Reza Farid Hosseini, Farahzad Jabbari Azad, Jason Schenkel, Abdolreza Varasteh, Mahmoud Mahmoudi Source Type: journals

Flow cytometry antibody screening using pooled red cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
For red cell alloantibody screening, the column agglutination technique (CAT) is used extensively, and flow cytometry (FC) screening has recently been demonstrated to be accurate, rapid, and cost effective. We attempted to determine whether the high sensitivity of FC allows pooling of screening red cells, which is generally not an acceptable technique in CAT.For FC screening, a commercial two-cell screening panel was utilized for the preparation of individual cells (CSi), as well as pooled cells diluted 1 in 2 (CSp), and 1 in 3 (CS1/3). Another panel was pooled from 120 randomly selected group O donors (RSp).Comparing the ...
Source: Cytometry Part B: Clinical Cytometry - August 26, 2009 Category: Molecular Biology Authors: Dong Il Won, Ok-Ju Jung, Yun Soo Lee, Sang Geol Kim, Jang Soo Suh Source Type: journals

Four- and five-color flow cytometry analysis of leukocyte differentiation pathways in normal bone marrow: A reference document based on a systematic approach by the GTLLF and GEILEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The development of multiparameter flow cytometry (FCM) and increasingly sophisticated analysis software has considerably improved the exploration of hematological disorders. These tools have been widely applied in leukaemias, lymphomas, and myelodysplasias, yet with very heterogeneous approaches. Consequently, there is no extensive reference document reporting on the characteristics of normal human bone marrow (BM) in multiparameter FCM. Here, we report a reference analysis procedure using relevant antibody combinations in normal human BM.A first panel of 23 antibodies, constructed after literature review, was tested in fo...
Source: Cytometry Part B: Clinical Cytometry - August 25, 2009 Category: Molecular Biology Authors: Christine Arnoulet, Marie C. Béné, Françoise Durrieu, Jean Feuillard, Chantal Fossat, Bernard Husson, Hélène Jouault, Marc Maynadié, Francis Lacombe Source Type: journals

One-tube HLA-B27/B2708 typing by flow cytometry using two "Anti-HLA-B27" monoclonal antibody reagentsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Conclusions: Our one-tube B27/B2708 assay is simple, robust, uses two "B27" moabs for typing precision and security, does not suffer from interference by HLA-B7 or other cross-reactive antigens and has the obvious advantage of using a single tube per typing. © 2009 Clinical Cytometry Society (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - August 19, 2009 Category: Molecular Biology Authors: Chris Darke, Ernest Coates Source Type: journals

Sample stability and variability of B-cell subsets in blood from healthy subjects and patients with systemic lupus erythematosusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study provides technical perspectives for those conducting immunophenotypic analyses of B-cells in patients with SLE. We envision that our data, which addresses sample stability issues and presents a method to describe the relative reliability of one measure over another, holds value for clinical assessments of B-cells in SLE and the evaluation of investigational agents designed to modify the B-cell compartment. © 2009 Clinical Cytometry Society (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - August 10, 2009 Category: Molecular Biology Authors: Shelley Sims Belouski, Daniel Wallace, Michael Weisman, Mariko Ishimori, Lisa Hendricks, Debra Zack, Mike Vincent, Erik Rasmussen, John Ferbas, James Chung Source Type: journals

QASI, an international quality management system for CD4 T-cell enumeration focused to make a global differenceEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A significant worldwide mobilization effort to treat people with HIV disease began in 2003. Most guidelines for initiating antiretroviral therapy require reliable and reproducible CD4 T-cell counting. Therefore, any effort that improves global availability of quality managed assessment schemes for CD4 T-cell enumeration is a positive achievement for the clinical management of AIDS on a worldwide scale.The Canadian QASI-Quality Management System (QMS) has been in operation for over a decade. More recently, QMS has fine-tuned its strategy to optimize its global impact in the fight against the HIV/AIDS pandemic. Three modific...
Source: Cytometry Part B: Clinical Cytometry - July 13, 2009 Category: Molecular Biology Authors: Michèle Bergeron, Tao Ding, Guy Houle, Linda Arès, Christian Chabot, Nadia Soucy, Peggy Seely, Alice Sherring, Dragica Bogdanovic, Sylvie Faucher, Randy Summers, Ray Somorjai, Paul Sandstrom Source Type: journals

Clinical evaluation of a simple image cytometer for CD4 enumeration on HIV-infected patientsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Affordable, easy-to-use, and reliable CD4+ T lymphocyte enumeration systems are needed in resource-constrained settings to monitor HIV.A simple image cytometer was used to count fluorescently labeled CD4+ T and CD8+ T lymphocytes from CD3 immunomagnetically selected cells on blood specimens of 460 HIV-1-infected patients in Siriraj Hospital, Bangkok, Thailand. Results were compared with flow cytometry (FCM).CD4+ T lymphocyte counts by image cytometer were comparable (R [ge] 0.97) with those by the FACSCount and the FACScan with a bias of 7.3 and 9.1%, respectively. At very low CD4+ T lymphocyte counts ([le]50/[mu]l) some o...
Source: Cytometry Part B: Clinical Cytometry - July 7, 2009 Category: Molecular Biology Authors: Xiao Li, Christian Breukers, Aurel Ymeti, Kovit Pattanapanyasat, Kasama Sukapirom, Leon W. M. M. Terstappen, Jan Greve Source Type: journals

Considerations for the control of background fluorescence in clinical flow cytometryEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This article focuses on three major causes of background (autofluorescence, spectral overlap, and undesirable antibody binding) by reviewing individual aspects of flow cytometric measurements that contribute to these causes. The appropriate use of controls facilitates a thorough understanding of these contributing factors as well as the development of robust cell labeling protocols intended for routine flow cytometric analysis. We present a set of recommendations that enables the user to develop an optimized cell labeling protocol that minimizes background and maximizes the ability to reliably distinguish between a positiv...
Source: Cytometry Part B: Clinical Cytometry - July 4, 2009 Category: Molecular Biology Authors: Ruud Hulspas, Maurice R.G. O'Gorman, Brent L. Wood, Jan W. Gratama, D. Robert Sutherland Source Type: journals

Harmonization of light scatter and fluorescence flow cytometry profiles obtained after staining peripheral blood leucocytes for cell surface-only versus intracellular antigens with the Fix & PermTM reagentEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Staining for intracellular markers with the Fix & PermTM reagent is associated with variations in the scatter properties of leucocytes, limiting automated analysis of flow cytometry (FCM) data. Here, we investigated those variables significantly contributing to changes in the light scatter, autofluorescence, and bcl2 staining characteristics of peripheral blood (PB) leucocytes, after fixation with Fix & PermTM. Our major aim was to evaluate a new mathematical approach for automated harmonization of FCM data from datafiles corresponding to aliquots of a sample treated with cell-surface-only versus Fix & Perm intracellular s...
Source: Cytometry Part B: Clinical Cytometry - July 1, 2009 Category: Molecular Biology Authors: Elaine Sobral da Costa, Rodrigo Tosta Peres, Julia Almeida, Quentin Lécrevisse, María Elena Arroyo, Cristina Teodósio, Carlos Eduardo Pedreira, Jacques J. M. van Dongen, Alberto Orfao, on behalf of the EuroFlow Consortium Source Type: journals

Advances in complex multiparameter flow cytometry technology: Applications in stem cell researchEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Flow cytometry and cell sorting are critical tools in stem cell research. Recent advances in flow cytometric hardware, reagents, and software have synergized to permit the stem cell biologist to more fully identify and isolate rare cells based on their immunofluorescent and light scatter characteristics. Some of these improvements include physically smaller air-cooled lasers, new designs in optics, new fluorescent conjugate-excitation pairs, and improved software to visualize data, all which combine to open up new horizons in the study of stem cells, by enhancing the resolution and specificity of inquiry. In this review, t...
Source: Cytometry Part B: Clinical Cytometry - June 5, 2009 Category: Molecular Biology Authors: Frederic Preffer, David Dombkowski Source Type: journals

Description of flow cytometry examinations related to human cell differentiation molecules in clinical immunologyEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Human cell differentiation molecules, also known as "CD", are cell-surface molecules reacting with different monoclonal antibodies. The fraction of cells belonging to each immunophenotype is measured by flow cytometry. This brief communication proposes the application into clinical immunological laboratories, of the syntax recommended by the International Union of Pure and Applied Chemistry and the International Federation of Clinical Chemistry and Laboratory Medicine to describe biological properties. Following this syntax, all properties related to human cell differentiation molecules examined by flow cytometry, but not ...
Source: Cytometry Part B: Clinical Cytometry - June 2, 2009 Category: Molecular Biology Authors: Xavier Fuentes-Arderiu, Mariona Mestre Source Type: journals

Correlation of flow cytometrically determined expression of ZAP-70 using the SBZAP antibody with IgVH mutation status and cytogenetics in 1,229 patients with chronic lymphocytic leukemiaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
ZAP-70 provides an important prognostic information in chronic lymphocytic leukemia (CLL); however, the most appropriate antibody clone and way of analysis have not yet been defined.We determined ZAP-70 expression in 1,229 patients with CLL using the SBZAP clone by applying three different ways of analysis (% positive B-cells, mean fluorescence intensity (MFI) in B-cells, MFI ratio T-cells:B-cells).ZAP-70 expression was related to somatic hypermutation status of IgVH genes for all three ways of analysis (P < 0.0001 each). The strongest correlation was found for MFI ratio (r = -0.475, P < 0.0001); cases with mutated and unm...
Source: Cytometry Part B: Clinical Cytometry - May 28, 2009 Category: Molecular Biology Authors: Wolfgang Kern, Frank Dicker, Susanne Schnittger, Claudia Haferlach, Torsten Haferlach Source Type: journals

Aberrant T-lymphocytes in refractory coeliac disease are not strictly confined to a small intestinal intraepithelial localizationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Refractory coeliac disease (RCD) is characterized by persisting mucosal pathology in spite of a strict gluten free diet (GFD). In RCD type II, phenotypically aberrant (CD7+CD3-CD4/8-cytoplasmicCD3+) T-lymphocytes are present within the intraepitelial lymphocyte (IEL) population in the small intestine, and 50-60% of these patients develops an enteropathy associated T-cell lymphoma (EATL).To investigate whether aberrant T-lymphocytes in RCD II can be detected in other parts of the small intestinal mucosa besides the intraepithelial compartment. Additionally, the presence of aberrant T-lymphocytes was analyzed in two RCD II p...
Source: Cytometry Part B: Clinical Cytometry - May 15, 2009 Category: Molecular Biology Authors: W.H.M. Verbeek, B.M.E. von Blomberg, V.M.H. Coupe, S. Daum, C.J.J. Mulder, M.W.J. Schreurs Source Type: journals

From research tool to routine test: CD38 monitoring in HIV patientsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
CD38 expression on CD8+ T lymphocytes in HIV-infected patients is monitored by flow cytometry (FCM). There is however no consensus re CD38 protocols, analyses or result reporting within/between laboratories. Internal quality control measures (QC) were established for a standardized CD38 protocol and a system proposed for reporting CD38 fluctuation in longitudinal HIV+ patient monitoring.A single-platform (SP) CD38/CD8 protocol was "piggy-backed" onto the standardized "panleucogating" CD45/CD4+ protocol. A weekly QC was established to monitor instrument stability (FlowSETTM) and absolute cell count accuracy and reproducibil...
Source: Cytometry Part B: Clinical Cytometry - May 7, 2009 Category: Molecular Biology Authors: Lindi M. Coetzee, Szun Szun Tay, Denise Lawrie, George Janossy, Deborah K. Glencross Source Type: journals

Flow cytometric evaluation of CD38 expression assists in distinguishing follicular hyperplasia from follicular lymphomaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study identified decreased CD38 as a common finding in FL in comparison with FH and provides an additional tool to help differentiate FL from FH by flow cytometry. © 2009 Clinical Cytometry Society (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - April 20, 2009 Category: Molecular Biology Authors: Kristin Mantei, Brent L. Wood Source Type: journals

CD22 expression on blastic plasmacytoid dendritic cell neoplasms and reactivity of anti-CD22 antibodies to peripheral blood dendritic cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We identified CD22 expression on a blastic plasmacytoid dendritic cell (pDC) neoplasm presenting as a leukemia in a child. CD22 expression, as determined by the antibody s-HCL-1, was also noted on the neoplastic cells from three additional patients with blastic pDC tumors identified at our institution. Subsequently we determined that peripheral blood pDCs react with the s-HCL-1 antibody demonstrating that normal pDCs express CD22. Evaluation of five additional anti-CD22 antibodies indicated that staining of pDCs with these reagents was poor except for s-HCL-1. Therefore, the detection of CD22 on pDCs is best demonstrated w...
Source: Cytometry Part B: Clinical Cytometry - April 20, 2009 Category: Molecular Biology Authors: Edmunds Z. Reineks, Ebenezer S Osei, Arlene Rosenberg, Jeffrey Auletta, Howard J. Meyerson Source Type: journals

A tag-less method of sorting stem cells from clinical specimens and separating mesenchymal from epithelial progenitor cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We present a tag-less, flow-assisted method to purify, distinguish, and sort pluri/multipotent SCs obtained from clinical specimens, based on differences in the biophysical properties that cells acquire when in suspension under fluidic conditions. A suspension of cells in a transport fluid is injected into a ribbon-like capillary device by continuous flow. In a relatively short time (about 30 min), sorted cells are collected.We obtained baseline separation between MSCs and epithelial cells, which are important contaminants of isolated MSCs. The extent of separation is evaluated by flow cytometry through detection of a spec...
Source: Cytometry Part B: Clinical Cytometry - April 18, 2009 Category: Molecular Biology Authors: Barbara Roda, Pierluigi Reschiglian, Andrea Zattoni, Francesco Alviano, Giacomo Lanzoni, Roberta Costa, Arianna Di Carlo, Cosetta Marchionni, Michele Franchina, Laura Bonsi, Gian Paolo Bagnara Source Type: journals

Predictive value of hematological and phenotypical parameters on postchemotherapy leukocyte recoveryEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Grade IV chemotherapy toxicity is defined as absolute neutrophil count 10; median 27.7), characterized a slow hematological recovery. MPXI levels were directly related to the cycle number and inversely related to the absolute number of LUCs and CD34+/CD45+ cells. A faster hematological recovery was associated with a higher LUC increase per day (0.56% vs. 0.25%), higher blast (median 36.7/[mu]L vs. 19.5/[mu]L) and CD34+/CD45+ cell (median 2.2/[mu]L vs. 0.82/[mu]L) counts.Our study showed that some biological indicators such as MPXI, LUCs, blasts, and CD34+/CD45+ cells may be of clinical relevance in predicting individual he...
Source: Cytometry Part B: Clinical Cytometry - April 8, 2009 Category: Molecular Biology Authors: A. Bononi, F. Lanza, L. Ferrari, M. Gusella, G. Gilli, V. Abbasciano, D. Campioni, A. Russo, D. Menon, F. Albertini, L. Stievano, C. Barile, G. Crepaldi, S. Toso, E. Ferrazzi, F. Pasini Source Type: journals

HLA-DRneg patients without acute promyelocytic leukemia show distinct immunophenotypic, genetic, molecular, and cytomorphologic characteristics compared to acute promyelocytic leukemiaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Loss of HLA-DR and CD34 is a well-known characteristic of malignant promyelocytes in acute promyelocytic leukemia (APL). However, this immunophenotype is not specific for APL. The purpose of this study was to investigate whether further biological characterization of the HLA-DRneg acute myeloid leukemia patients would allow more clearly define criteria to separate APL from non-APL patients.Immunophenotyping, cytogenetics, molecular analyses, and cytomorphology were prospectively performed within routine leukemia diagnostics of 800 patients included in different prospective acute myeloid leukemia multicenter trials.Beside 6...
Source: Cytometry Part B: Clinical Cytometry - March 18, 2009 Category: Molecular Biology Authors: Uta Oelschlaegel, Brigitte Mohr, Markus Schaich, Ulrike Schäkel, Frank Kroschinsky, Thomas Illmer, Gerhard Ehninger, Christian Thiede Source Type: journals

Phagocytosis and postphagocytic reaction of cord blood and adult blood monocyte after infection with green fluorescent protein-labeled Escherichia coli and group B StreptococciEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Neonatal sepsis is characterized by an excessive inflammatory response induced by immune cells (monocytes). We investigated the initial stage of monocyte-pathogen interaction, i.e. bacterial ingestion and degradation at the single-cell level, by comparing a new flow cytometric procedure with culture methods. We also examined the hypothesis that, in terms of phagocytosis-induced cell death (PICD), phenotype, or cytokine production, cord blood monocytes (CBMO) differ from monocytes derived from adults (peripheral blood monocytes, PBMO).Phagocytosis and intracellular degradation were assessed by means of flow cytometry and ba...
Source: Cytometry Part B: Clinical Cytometry - March 16, 2009 Category: Molecular Biology Authors: Ch. Gille, A. Leiber, I. Mundle, B. Spring, H. Abele, B. Spellerberg, H. Hartmann, Ch. F. Poets, Th. W. Orlikowsky Source Type: journals

A rapid flow cytometric screening test for X-linked lymphoproliferative disease due to XIAP deficiencyEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Deficiency of X-linked inhibitor of apoptosis (XIAP), caused by BIRC4 gene mutations, is the second known cause of X-linked lymphoproliferative disease (XLP), a rare primary immunodeficiency that often presents with life-threatening hemophagocytic lymphohistiocytosis (HLH). Rapid diagnosis of the known genetic causes of HLH, including XIAP deficiency, facilitates the initiation of life-saving treatment and preparation for allogeneic hematopoietic cell transplantation (HCT). Until now, a rapid screening test for XIAP deficiency has not been available.To develop a flow cytometric screening test for XIAP deficiency, we first ...
Source: Cytometry Part B: Clinical Cytometry - March 13, 2009 Category: Molecular Biology Authors: Rebecca A. Marsh, Joyce Villanueva, Kejian Zhang, Andrew L. Snow, Helen C. Su, Lisa Madden, Rajen Mody, Brenda Kitchen, Dan Marmer, Michael B. Jordan, Kimberly A. Risma, Alexandra H. Filipovich, Jack J. Bleesing Source Type: journals

Monocytes and T lymphocytes contribute to a predominance of interleukin 6 and interleukin 10 in systemic lupus erythematosusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
To investigate the contribution of T lymphocytes and monocytes to cytokine production in systemic lupus erythematosus (SLE).Forty-five SLE patients and 19 healthy volunteers were included. Serum levels of tumor necrosis factor alpha (TNF[alpha]), interferon gamma (IFN[gamma]), interleukin (IL)-6, and IL10 were quantified by ELISA. The cytokine production capacities of peripheral blood mononuclear cells were assessed by culturing in vitro with PMA+Ionomycin or LPS. The intracellular cytokine expression was measured by flow cytometry in T lymphocytes and monocytes, respectively. The influence of the disease activity (measure...
Source: Cytometry Part B: Clinical Cytometry - February 8, 2009 Category: Molecular Biology Authors: Susana Mellor-Pita, Maria J. Citores, Raquel Castejon, Miguel Yebra-Bango, Pablo Tutor-Ureta, Silvia Rosado, Jose L. Andreu, Juan A. Vargas Source Type: journals

Absolute CD4+ cell count using a plastic microchip and a microscopic cell counterEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We have designed and evaluated the performance of a simple, rapid, and affordable method for counting CD4+ T-cells with the use of plastic microchips. This new system is an adaptation of a "no-lyse, no-wash," volumetric single platform assay, and absolute CD4+ counts are determined with the use of a microscopic scanning cell counter. To assess the CD4+ count test precision and linearity of the system, measured CD4+ counts were compared with two other reference assays (single and dual platform flow cytometry) with the use of 123 clinical samples including samples obtained from 35 HIV-infected patients, and artificially dilu...
Source: Cytometry Part B: Clinical Cytometry - January 8, 2009 Category: Molecular Biology Authors: Sook Young Bae, Hae Chul Park, Jun Seo Oh, Soo-Young Yoon, Dae Won Park, In Keun Choi, Hae Joong Kim, Jong Hyun Oh, Dae Sung Hur, Chanil Chung, Jun Keun Chang, J. Paul Robinson, Chae Seung Lim Source Type: journals

Close association of CD8+/CD38bright with HIV-1 replication and complex relationship with CD4+ T-cell countEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Measuring lymphocyte activation provides information in addition to CD4+ T-cell count for immune monitoring of HIV-1 infected patients. CD38 is a well-established activation marker that is generally analyzed on the whole population of CD8+ T-cells. Focusing specifically on CD38 high expression (CD8+/CD38bright) may be an interesting surrogate gating strategy because CD38bright characterizes principally activated memory cells.CD8+/CD38bright was investigated in 1,353 HIV-1 infected patients over a one-year period to establish relevant cutoff values and clarify the relationships of this marker with HIV-1 RNA viral load (VL) ...
Source: Cytometry Part B: Clinical Cytometry - December 13, 2008 Category: Molecular Biology Authors: Edouard Tuaillon, Yassine Al Tabaa, Vincent Baillat, Michel Segondy, Marie-Christine Picot, Jacques Reynes, Jean-Pierre Vendrell Source Type: journals

Mantle cell lymphoma with flow cytometric evidence of clonal plasmacytic differentiation: A case reportEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Plasmacytic differentiation in mantle cell lymphoma (MCL) occurs rarely. However, no flow cytometric studies that demonstrate plasmacytic (PC) differentiation in MCL have been reported. Herein, we report a case of MCL with PC differentiation identified by flow cytometry.Morphologic review was performed by hematoxylin and eosin (H&E) stained sections from paraffin-embedded lymph node, colon and bone marrow specimens, and Wright-Geimsa stained bone marrow aspirate smears and touch imprints. Immunohistochemical stains using antibodies against CD3, CD5, CD20, and cyclin-D1, and in-situ hybridization for kappa and lambda light ...
Source: Cytometry Part B: Clinical Cytometry - December 6, 2008 Category: Molecular Biology Authors: Hina Naushad, William W. L. Choi, Cynthia J. Page, Warren G. Sanger, Dennis D. Weisenburger, Patricia Aoun Source Type: journals

Proof of principle: An HIV p24 microsphere immunoassay with potential application to HIV clinical diagnosisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The measurement of CD4 counts and viral loads on a single instrument such as an affordable flow cytometer could considerably reduce the cost related to the follow-up of antiretroviral therapy in resource-poor settings. The aim of this study was to assess whether the HIV-1 p24 antigen could be measured using a microsphere-based flow cytometric (FC) assay and the experimental conditions necessary for processing plasma samples. A commercial anti-p24 antibody pair from Biomaric was used to develop a p24 microsphere immunoassay (MIA) using HIV culture supernatant as the source of antigen. The ultrasensitive Perkin Elmer enzyme ...
Source: Cytometry Part B: Clinical Cytometry - November 15, 2008 Category: Molecular Biology Authors: Pascale Ondoa, Chris Vereecken, Eugene Lekeawung Asahchop, Amber Litzroth, Aziz Diallo, Katrien Fransen, Tandakha Dieye, Robin Ryder, Souleymane Mboup, Luc Kestens Source Type: journals

The absolute counting of red cell-derived microparticles with red cell bead by flow rate based assayEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study demonstrates that FCB is suitable and more affordable for RMPs quantitation in the clinical samples. This method is a low cost and interchangeable to latex bead-based method for generating the absolute counts in the resource-limited areas. © 2008 Clinical Cytometry Society (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - November 7, 2008 Category: Molecular Biology Authors: Duangdao Nantakomol, Malika Imwong, Ingfar Soontarawirat, Duangporn Kotjanya, Chulalak Khakhai, Jun Ohashi, Pornlada Nuchnoi Source Type: journals

A decreased positivity for CD90 on human mesenchymal stromal cells (MSCs) is associated with a loss of immunosuppressive activity by MSCsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, we investigated the basis of immunomodulant possible variability using hMSC from different sources (amniotic membrane, chorion, and bone marrow from either healthy subjects or patients with hematological malignancies, HM) and having discordant positivity for several immunological markers. The CD90+ hMSC reduced lymphoproliferative response in phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMC) via sHLA-G and IL-10 up-modulation. On the contrary, hMSC showing a significantly lower expression for CD90 antigen, elicited a lymphoproliferative allogeneic response in PHA/PBMCs without any ...
Source: Cytometry Part B: Clinical Cytometry - November 4, 2008 Category: Molecular Biology Authors: Diana Campioni, Roberta Rizzo, Marina Stignani, Loredana Melchiorri, Luisa Ferrari, Sabrina Moretti, Antonio Russo, Gian Paolo Bagnara, Laura Bonsi, Francesco Alviano, Giacomo Lanzoni, Antonio Cuneo, Olavio R. Baricordi, Francesco Lanza Source Type: journals

ErratumEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
No abstract. (Source: Cytometry Part B: Clinical Cytometry)
Source: Cytometry Part B: Clinical Cytometry - October 31, 2008 Category: Molecular Biology Authors: Francis Mandy Source Type: journals

Overexpression of CD7 in classical Hodgkin lymphoma-infiltrating T lymphocytesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study characterizes the FC immunophenotype of these T lymphocytes to determine whether they can be used to assist in the diagnosis of HL.Cell suspensions from 76 lymph nodes involved by HL and 156 lymph nodes with reactive lymphadenopathy (LAD) were analyzed by flow cytometry to assess the expression of T-cell antigens.The CD4:CD8 ratio and CD7 expression in both CD4(+) and CD8(+) T cells are increased in HL compared with reactive lymph nodes and there are significant differences between these features in different subtypes of HL. However, only the expression of CD7 in CD4(+) T cells distinguishes between HL and react...
Source: Cytometry Part B: Clinical Cytometry - October 29, 2008 Category: Molecular Biology Authors: Adam C. Seegmiller, Nitin J. Karandikar, Steven H. Kroft, Robert W. McKenna, Yin Xu Source Type: journals

Peripheral blood flow cytometry based diagnosis in primary cutaneous aggressive epidermotropic CD8+ T-cell lymphoma: Implications for more extensive useEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Primary cutaneous epidermotropic CD8-positive T-cell lymphoma represents an aggressive form of T-cell cutaneous lymphomas. Diagnosis is based on biopsies obtained from skin lesions. Here, we would like to report a case diagnosed by using flow cytometry performed on peripheral blood mononuclear cells. Moreover, an important finding was the difference in the results on targeting the CD8 antigen by using two different commercially available monoclonal antibodies. Perhaps, more than one antibody should be used in primary cutaneous aggressive epidermotropic CD8-positive T-cell lymphomas, in orderto be more accurate in the diagn...
Source: Cytometry Part B: Clinical Cytometry - October 27, 2008 Category: Molecular Biology Authors: Leonidas Benetatos, Lefkothea Dova, Gerasimos Baxevanos, Nikolaos I. Kolaitis, Konstantinos L. Bourantas, Georgios Vartholomatos Source Type: journals

Flow cytometric differential of leukocyte populations in normal bone marrow: Influence of peripheral blood contamination1Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Availability of immunophenotypic reference values for the various leukocyte populations distributed in bone marrow may be helpful to recognize abnormal bone marrow development and, therefore, useful as first screening of individuals with suspected hematological malignancies or other hematopoietic disorders.A single tube four-color staining panel (CD66abce/CD14/CD45/CD34) together with a predefined gating strategy was utilized to immunologically differentiate the distribution of the major leukocyte populations in bone marrow aspirates of healthy donors. The sample-blood erythrocyte ratio was applied to assess the amount of ...
Source: Cytometry Part B: Clinical Cytometry - October 22, 2008 Category: Molecular Biology Authors: R. A. Brooimans, J. Kraan, W. van Putten, J. J. Cornelissen, B. Löwenberg, J. W. Gratama Source Type: journals

A comparison of the choice of monoclonal antibodies for recovery of fetal cells from maternal blood using FACS for noninvasive prenatal diagnosis of hemoglobinopathiesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Isolation of adequate numbers of fetal cells circulating in the maternal circulation is the major hurdle in developing noninvasive prenatal diagnostic procedures. We used flow cytometry and a combination of different monoclonal antibodies to compare the yield and purity of the fetal nucleated red blood cells at different periods of gestation.Using a Percoll discontinuous gradient, the fetal nucleated erythrocytes were enriched from 7 ml maternal blood. In 100 samples, the enriched cells were stained with CD45, anti-fetal hemoglobin, and glycophorin A antibodies and in 30 samples they were stained with CD45, anti-fetal hemo...
Source: Cytometry Part B: Clinical Cytometry - October 2, 2008 Category: Molecular Biology Authors: Edna D'Souza, Kanjaksha Ghosh, Roshan Colah Source Type: journals

Overexpression of CD49f in precursor B-cell acute lymphoblastic leukemia: Potential usefulness in minimal residual disease detectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The persistence of minimal residual disease (MRD) following therapy is an established prognostic factor in precursor B-cell acute lymphoblastic leukemia (pB-ALL). Detection of MRD in pB-ALL by flow cytometric immunophenotyping requires demonstration of abnormal antigen expression in leukemic B-cell precursors relative to that of normal B-cell precursors. The gene encoding CD49f (integrin [alpha]-6) is one of several whose overexpression in pB-ALL at diagnosis has been associated with the subsequent detection of MRD. However, whether CD49f might be a useful reagent in the immunophenotypic detection of MRD in pB-ALL has not ...
Source: Cytometry Part B: Clinical Cytometry - October 2, 2008 Category: Molecular Biology Authors: Joseph A. DiGiuseppe, Sheila G. Fuller, Michael J. Borowitz Source Type: journals

A comparison of multiplex suspension array large-panel kits for profiling cytokines and chemokines in rheumatoid arthritis patientsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study evaluated four multiplex kits (Bio-Plex, LINCOplex, Fluorokine, and Beadlyte) that contained 27, 29, 20, and 22 analytes each, respectively, for the analysis of immunomodulators in plasma of patients with rheumatoid arthritis (RA) who underwent treatment with antibody against CD20 (rituximab), a B-cell reductive therapy.Multiplex kits were tested on serial plasma samples obtained from six RA patients at baseline and multiple time points (3, 6, and 9 months) post-treatment with rituximab. The RA patients included in this study had previously failed therapy with disease modifying anti-arthritis drugs (DMARD) and t...
Source: Cytometry Part B: Clinical Cytometry - September 30, 2008 Category: Molecular Biology Authors: Imran H. Khan, V. V. Krishnan, Melanie Ziman, Kim Janatpour, Ted Wun, Paul A. Luciw, Joseph Tuscano Source Type: journals

CD4 and CD8 enumeration for HIV monitoring in resource-constrained settingsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We developed a volumetric single platform image cytometer (SP ICM) that is dedicated to count CD4+ and CD8+ T lymphocytes for HIV monitoring in resource-constrained settings. The instrument was designed to be low-cost, yet reliable, easy-to-use, and robust.Whole blood is incubated with CD3-magnetic nanoparticles, CD4-phycoerythrin (PE), and CD8-peridinin-chlorophyll-protein complex (PerCP). The CD3 cells are immunomagnetically attracted to an analysis surface, where fluorescence images of CD4+ and CD8+ T lymphocytes are recorded and analyzed, respectively. We compared CD4, CD8 counts, and CD4/CD8 ratio obtained by the SP I...
Source: Cytometry Part B: Clinical Cytometry - September 29, 2008 Category: Molecular Biology Authors: Xiao Li, Christian Breukers, Aurel Ymeti, Björn Lunter, Leon W. M. M. Terstappen, Jan Greve Source Type: journals

A rapid, fluorescence-based assay for detecting antigenic modulation of the acetylcholine receptor on human cell linesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Myasthenia gravis (MG) is an autoimmune disease affecting [sim]40,000 patients in the United States. One of the major mechanisms of disease pathology in MG is the binding, internalization, and eventual destruction of acetylcholine receptors (AChR) at the neuromuscular junction by cross-linking AChR-specific autoantibodies. This process, known as antigenic modulation, ultimately attenuates the ability of muscle cells to contract in response to signals from neurons, leading to muscle weakness and fatigue. For this reason, antigenic modulation of the AChR on cultured cells has become an important diagnostic tool for assessing...
Source: Cytometry Part B: Clinical Cytometry - September 29, 2008 Category: Molecular Biology Authors: Dennis Keefe, Donna Hess, Jennifer Bosco, Socrates Tzartos, Jan Powell, Justin Lamsa, Serene Josiah Source Type: journals

Assessment of mitochondrial toxicity by analysis of mitochondrial protein expression in mononuclear cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Real-time PCR has quantified decreased mitochondrial DNA levels in association with nucleoside reverse transcriptase inhibitor (NRTI) therapy of HIV-infected populations. However, real-time PCR is best suited to distinguish log differences in an analyte. In an effort to monitor individuals in more detail, we developed a flow cytometric assay to gauge mitochondrial function.Flow cytometric quantification of a mitochondrial DNA-encoded mitochondrial protein (cytochrome c oxidase subunit I (COX-I)) and a nuclear DNA-encoded mitochondrial protein [ATP synthase subunit D (Sub-D)] was optimized and validated.Intra-assay and inte...
Source: Cytometry Part B: Clinical Cytometry - September 29, 2008 Category: Molecular Biology Authors: Chen-Han Lin, Derek D. Sloan, Chung H. Dang, Thor Wagner, Albert J. E. Cabrera, Nicole H. Tobin, Lisa M. Frenkel, Keith R. Jerome Source Type: journals

Cytomics and nanobioengineeringEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The finding that an individual's genome differs as much as by many million variants from that of the human reference assembly diminished the great enthusiasm that every disease could be predicted based on nucleotide polymorphisms. Even individual cells of an organ may be specifically equipped to perform specific tasks and that the information of individual cells in a cell system is key information to understand function or dysfunction. Therefore, cytomics received great attention during the last years as it allows to quantitatively and qualitatively analyzing great number of individual cells, cell constituents, and of thei...
Source: Cytometry Part B: Clinical Cytometry - September 25, 2008 Category: Molecular Biology Authors: Arkadiusz Pierzchalski, Andrea Robitzki, Anja Mittag, Frank Emmrich, Ulrich Sack, José-Enrique O'Connor, József Bocsi, Attila Tárnok Source Type: journals

Flow cytometric quantitation of natural killer cells and T lymphocytes expressing T-cell receptors alpha/beta and gamma/delta is not helpful in distinguishing benign from malignant body cavity effusionsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Quantitation of natural killer (NK) cells in benign and malignant effusions has yielded conflicting results in the past. Studies have claimed higher, lower, and essentially equal percentages of NK cells for benign and malignant effusions. In addition, virtually no literature exists on the numbers and distribution of T lymphocytes expressing T-cell receptor alpha/beta (TCR [alpha]/[beta]) and T-cell receptor gamma/delta (TCR [gamma]/[delta]) in body effusions.Using multicolor flow cytometry and sequential gating techniques, NK cells and T lymphocytes expressing TCR [alpha]/[beta] and TCR [gamma]/[delta] were identified and ...
Source: Cytometry Part B: Clinical Cytometry - September 20, 2008 Category: Molecular Biology Authors: Dennis B. Cornfield, Shereen M. F. Gheith Source Type: journals

Immunophenotypic profile of acute leukemia: Critical analysis and insights gained at a tertiary care center in IndiaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
To analyze the spectrum of various types and subtypes of acute leukemia.Two thousand five hundred and eleven consecutive new referral cases of acute leukemia (AL) were evaluated based on WHO classification.It included 1,471 cases (58%) of acute lymphoblastic leukemia (ALL), 964 cases (38%) of acute myeloid leukemia (AML), 45 cases (1.8%) of chronic myelogenous leukemia in blast crisis (CMLBC), 37 cases (1.5%) of biphenotypic acute leukemia (BAL), 1 case of Triphenotypic AL, and 2 cases of acute undifferentiated leukemia (AUL). Common subtypes of ALL were B-cell ALL (76%), which comprised of intermediate stage/CALLA positiv...
Source: Cytometry Part B: Clinical Cytometry - September 19, 2008 Category: Molecular Biology Authors: S. Gujral, Y. Badrinath, A. Kumar, P. G. Subramanian, G. Raje, H. Jain, A. Pais, P. S. Amre Kadam, S. D. Banavali, B. Arora, P. Kumar, V. G. Hari Menon, P. A. Kurkure, P. M. Parikh, S. Mahadik, A. B. Chogule, S. C. Shinde, C. N. Nair Source Type: journals