scholarly journals The use of a hematology analyzer with a new generation of software as an alternative to flow cytometry for enumerating residual white blood cells in blood components

Transfusion ◽  
2019 ◽  
Vol 60 (1) ◽  
pp. 155-164 ◽  
Author(s):  
Richard Alejo Blanco ◽  
Chloe Cavagnetto ◽  
Laura Willmott ◽  
Elif Aydogdu ◽  
Nicola Akinyemi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
White Blood Cells ◽  
Blood Components ◽  
Hematology Analyzer ◽  
New Generation

scholarly journals FLOW CYTOMETRY FOR MEASURING RESIDUAL LEUKOCYTES IN BLOOD PLASMA

2019 ◽  
Vol 64 (1) ◽  
pp. 66-72
Author(s):  
V. S. Kozyreva ◽  
A. N. Shilova ◽  
O. S. Shkoda
Keyword(s):  
Blood Plasma ◽  
Blood Cells ◽  
White Blood Cells ◽  
Flow Cytometer ◽  
Blood Components ◽  
Blood Products ◽  
Transfusion Therapy ◽  
Analytical Instruments ◽  
Hematology Analyzer ◽  
In Blood Plasma

Background. Immunological and infectious safety of blood components is considered to be a key condition for high-quality transfusion therapy. The progress of blood filtration methods contributes to the development of methods for the validation of residual leukocyte control in blood products.The aim of the study was to compare the efficiency of flow cytometer with hematology analyzer for counting residual leukocytes in blood productsMethods. In this study, we compared the efficiency of two analytical instruments for counting residual leukocytes in 191 blood plasma samples: a flow cytometer (Navios Beckman Coulter) with a LeukoSure commercial reagent kit and a Sysmex XT4000i hematology analyzer.Results. All the samples under investigation met technical regulation requirements. Most of the samples were characterized by a significantly low amount of residual leukocytes, which fact presented a particular scientific interest. The use of a Navios Beckman Coulter flow cytometer and a commercial reagent kit allowed us to detect 15 or lower cells per μL in 83 % of the samples. The use of a Sysmex XT- 4000i automatic hematology analyzer for the calculation of residual white blood cells in this range proved to be limited due to the sensitivity of the device and the absence of certified reference materials with a low white blood cell count.Conclusion. Our results show that a Navios Beckman Coulter flow cytometer with a commercial reagent kit is suitable for measuring residual leukocytes in blood plasma. This instrument is most appropriate for samples with a minimum number of cells. A significant variability of the amount of residual white blood cells in blood plasma confirms the importance of inspecting the content of leukocytes in all blood components.

Direct freezing of whole blood enables analysis of leucocyte markers by flow cytometry: a proof-of-concept study

2021 ◽  
Author(s):  
Pénélope Bourgoin ◽  
Inès Ait Belkacem ◽  
Isabelle Arnoux ◽  
Pierre-Emmanuel Morange ◽  
Fabrice Malergue
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Proof Of Concept ◽  
Fresh Blood ◽  
Blood Samples ◽  
Step Flow ◽  
Freezing Method ◽  
One Step

Aim: A new one-step flow cytometry procedure has been recently demonstrated for identifying subjects with infections, but only for fresh whole blood samples. The goal of this study was to assess its applicability on frozen samples, by proposing a new method to perform the sample freezing directly and easily. Methods: Fresh blood was tested, then frozen either directly or with dimethylsulfoxide and serum. Common markers of white blood cells as well as infection-related biomarkers were tested. Results: All percentages of leucocyte subsets and levels of infection-related biomarkers were significantly correlated between frozen and fresh samples. Conclusion: The direct freezing method enables an accurate assessment of common cellular sub-populations and of levels of important infectious biomarkers via flow cytometry.

Distribution of dipyridamole in blood components among post-stroke patients treated with extended release formulation

2009 ◽  
Vol 102 (09) ◽  
pp. 538-543 ◽  
Author(s):  
Elena Sabaeva ◽  
Christopher Booze ◽  
Oliver D. Atar ◽  
Christian Eisert ◽  
Dan Hanley ◽  
...  
Keyword(s):  
Ischaemic Stroke ◽  
Blood Cells ◽  
Extended Release ◽  
White Blood Cells ◽  
Background Level ◽  
Blood Components ◽  
Stroke Patients ◽  
Antithrombotic Agent ◽  
Release Formulation ◽  
Post Stroke

SummaryExtended release dipyridamole (ERD) is widely used in patients after ischaemic stroke; however, the ability of this antithrombotic agent to be stored in different blood cells has never been explored in post-stroke patients. We hypothesised that since ERD is known to be highly lipophilic, the drug may be present not only in plasma, but also accumulated in platelets, leukocytes, and erythrocytes. Fifteen patients after documented ischaemic stroke were treated with Aggrenox (ERD and lowdose aspirin combination) BID for 30 days, and 12 of them completed the study. ERD concentrations in blood cells and platelet-poor plasma were measured by spectrofluorimetry at Baseline, Day 14, and Day 30 after the initiation of therapy. The background level of spectrofluorometry readings differs slightly among the blood components (132–211 ng/ml) due to the differences in the preparation of samples and cell isolation techniques. As expected, two weeks of ERD therapy produced steady-state plasma concentration of dipyridamole already at Day 14 (1,680 ±542 ng/ ml), followed by a slight not significant decrease at one month (1,619 ±408 ng/ml). Two weeks of therapy was sufficient to achieve a consistent dipyridamole accumulation in erythrocytes (361 ±43 ng/ml), but not in platelets (244 ±78 ng/ml), or leukocytes (275 ±49 ng/ml).In fact, white blood cells continued dipyridamole intake beyond 14 days period, and this increase (398 ± 66 ng/ml) was significant (p = 0.02) at 30 days. Treatment with ERD in post-stroke patients resulted not only in achievement of therapeutic plasma dipyridamole concentrations, but also deposition of the drug in erythrocytes and leukocytes, but not in platelets. If confirmed, these data will affect our better understanding of dipyridamole pleiotropy, and may explain long-term benefit of ERD formulation.

Expression of NTB-A in Normal Tissue, Non-Hodgkin Lymphomas and Investigation of Its Potential Therapeutic Impact for Chronic Lymphocytic Leukemia.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4741-4741
Author(s):  
Xiaoxian Zhao ◽  
Wouter Korver ◽  
Nichole Prescott ◽  
Arie Abo ◽  
Eric Hsi
Keyword(s):  
Flow Cytometry ◽  
B Cells ◽  
B Cell ◽  
Progenitor Cells ◽  
Blood Cells ◽  
Cell Lymphoma ◽  
White Blood Cells ◽  
B Cell Lymphoma ◽  
Hematologic Malignancies ◽  
Normal Tissues

Abstract Introduction: Expressed on T, B, NK cells and neutrophils, NTB-A belongs to the signaling lymphocytic activation molecule (SLAM) family of immune receptors. It was reported recently that cross-linking of NTB-A induces T cell activation and Ly108 (the presumed mouse orthologue of NTB-A) controls the production of reactive oxygen species in neutrophils. To date, little is known about its expression at the protein level in normal tissues, or in hematologic malignancies. Methods: We have generated monoclonal antibodies (mAbs) to NTB-A and used tissue microarrays (TMAs) to screen normal tissues and lymphomas. Western blotting and flow cytometry were used for confirmation of selected entities. Complement dependent cytotoxicity (CDC) assays were performed using the CellTiter-Glo Luminescent Cell Viability Assay kit (Promega). Results: NTB-A was not detected in normal tissues including heart, liver, breast, kidney, brain, lung, uterus, small intestine, skin, prostate, pancreas, ovary, bladder, testis and stomach (5/5 negative cases for all above tissues) by IHC. Expression was observed in lymphocytes of normal tonsil and spleen. In lymphomas, we found NTB-A expression in diffuse large B-cell lymphoma (DLBL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma and Burkitt lymphoma. Western blotting and flow cytometry of B-cell lymphoma cell lines DOHH2 (DLBL) and Raji (Burkitt) confirmed NTB-A expression in these lines. Myeloma cell lines were negative for expression. Flow cytometry of normal blood cells showed expression of NTB-A in B-cells, T-cells and NK cells (95.5 ± 6.7%, 62.6 ± 29.5% and 72.7 ± 26.5%, respectively) but not in CD66+ granulocytes. Furthermore, CD34+ progenitor cells from bone marrow were negative. Because of the expression in SLL, we focused on chronic lymphocytic leykemia (CLL) cells. All (15/15) patient samples were confirmed positive by flow cytometry. No significant differences in expression levels were observed comparing CLL B-cells vs normal B-cells (n = 10). However, anti-NTB-A antibodies were capable of inducing CDC in white blood cells from CLL patients (n=5) but not in normal white blood cells (n=5), in addition to CDC activity against Raji cells. Preliminary data also suggests that the CDC effect is greater in purified CLL B-cells than purified normal B-cells. Conclusions: These results show that NTB-A is expressed in normal lymphocytes but not in other solid tissues or in CD34+ progenitor cells. B cell malignancies, including Non-Hodgkin lymphoma cells appear to express NTB-A. In particular CLL patient cells express NTB-A, and an anti-NTB-A mouse mAb is capable of inducing CDC, suggesting NTB-A may be a potential immunotherapeutic target. Further studies examining the expression patterns in hematologic malignancies are ongoing.

Enumeration of residual white blood cells in leukoreduced blood products: Comparing flow cytometry with a portable microscopic cell counter

2016 ◽  
Vol 54 (2) ◽  
pp. 266-270 ◽  
Author(s):  
Silvia Castegnaro ◽  
Patrizia Dragone ◽  
Katia Chieregato ◽  
Alberta Alghisi ◽  
Francesco Rodeghiero ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
White Blood Cells ◽  
Blood Products ◽  
Cell Counter

scholarly journals Clinical chorioamnionitis at term VII: the amniotic fluid cellular immune response

2017 ◽  
Vol 45 (5) ◽  
Author(s):  
Alicia Martinez-Varea ◽  
Roberto Romero ◽  
Yi Xu ◽  
Derek Miller ◽  
Ahmed I. Ahmed ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Amniotic Fluid ◽  
Blood Cells ◽  
White Blood Cells ◽  
Flow Cytometric Analysis ◽  
Cytokine Expression ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Absolute ◽  
Cultivation Techniques ◽  
Spearman’S Correlation

AbstractObjectives:1) To characterize the cellular composition of the amniotic fluid of patients diagnosed with clinical chorioamnionitis at term, as a function of the presence or absence of microorganisms determined by cultivation techniques, and 2) to characterize the cytokine production by white blood cells present in the amniotic fluid using flow cytometry-based techniques.Materials and methods:Amniotic fluid samples from 20 women who had the diagnosis of clinical chorioamnionitis at term were analyzed using cultivation techniques (for aerobic and anaerobic bacteria as well as genital Mycoplasmas). Amniotic fluid IL-6 concentrations were determined by an enzyme-linked immunosorbent assay. Amniotic fluid leukocytes were visualized by using hematoxylin and eosin staining and immunofluorescence. Immunophenotyping of surface markers and cytokines was performed in amniotic fluid leukocytes using flow cytometry.Results:1) Neutrophils (CD45+CD15+ cells) were the most common leukocyte subset found in the amniotic fluid, followed by monocytes (CD45+CD14+ cells); other white blood cells (such as lymphocytes and natural killer cells) were scarce in the amniotic fluid; 2) the absolute counts of neutrophils and monocytes were significantly higher in patients with microorganisms found in the amniotic fluid than in those without detectable microorganisms, using cultivation techniques; 3) there was a significant correlation between the absolute counts of neutrophils and monocytes determined by flow cytometry (Spearman’s correlation=0.97; P<0.001); 4) there was a significant correlation between the absolute white blood cell count determined with a hemocytometer chamber and by flow cytometric analysis (Spearman’s correlation=0.88; P<0.001); and 5) the profile of cytokine expression differed between monocytes and neutrophils; while neutrophils predominantly produced TNF-α and MIP-1β, monocytes expressed higher levels of IL-1β and IL-1α.Conclusion:Flow cytometry analysis of the amniotic fluid of patients with intra-amniotic infection and clinical chorioamnionitis at term demonstrated that neutrophils and monocytes are the most common cells participating in the inflammatory process. We have characterized, for the first time, the differential cytokine expression by these cells in this important complication of pregnancy.

scholarly journals The utility of basic blood counts, WBC histogram and C-reactive protein in detecting malaria

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jun Nishimura ◽  
Parag Dharap ◽  
Sebastien Raimbault
Keyword(s):  
Blood Cells ◽  
Malaria Infection ◽  
Monsoon Season ◽  
Characteristic Curve ◽  
White Blood Cells ◽  
C Reactive Protein ◽  
Control Groups ◽  
Reactive Protein ◽  
Hematology Analyzer ◽  
And Control

Abstract Background Hematology analyzers display abnormal parameters during malaria infection providing insightful information for suspecting and assessing malaria infection. The goal of this study is to demonstrate the potential of a three-part differential hematology analyzer to assess malaria, provide information about the parasitemia, and discuss the importance of combining C-reactive protein (CRP) with hematology parameters to obtain further information about the malaria infection. Methods The present study shows the results of a case–control study during the monsoon season of years 2018 and 2019 in Mumbai, India. The study considers 1008 non-malaria febrile cases, 209 P. vivax and 31 P. falciparum positive malaria samples, five cases of mixed P. vivax and P. falciparum infection, and three co-infection cases of P. vivax and dengue. Raw data from the three-part analyzer LC-667G CRP (HORIBA) and the corresponding microscopic findings (golden standard for diagnosis of malaria) were obtained for each sample. Results The medians of platelet counts (PLT) were 102.5, 109.0, and 223.0 × 103/µL, while CRP medians were 67.4, 81.4 and 10.4 mg/L in P. vivax, P. falciparum and control groups respectively (p < 0.001 in Mann–Whitney U tests between malaria and control groups). Compared with negative samples, platelets counting less than 161.5 × 103/µL were observed on malaria patients (OR 19.12, 95% CI 11.89–30.75). Especially in P. vivax cases, an abnormal peak was frequently observed in the white blood cells (WBC) histogram around the 37fL channel. The events counted around that channel showed a linear correlation with the counting of red blood cells infected predominantly with larger parasitic forms. Parameters like CRP (rs = 0.325, p < 0.001), WBC (rs = 0.285, p < 0.001) and PLT (rs = − 0.303, p < 0.001) were correlated with the parasitemia of P. vivax samples. Between the malaria and dengue groups, the highest area under the receiver operating characteristic curve was observed on CRP (0.867, CRP ≥ 26.85 mg/L). Conclusions A three-part differential hematology analyzer has the potential to not only trigger malaria diagnosis confirmation but also assess the severity of the infection when CRP is considered.

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