scholarly journals EDTA Blood Cell Fraction

2020 ◽  
Author(s):  
Keyword(s):  
Blood Cell ◽  
Cell Fraction ◽  
Edta Blood

scholarly journals Small Red Blood Cell Fraction on the UF-1000i Urine Analyzer as a Screening Tool to Detect Dysmorphic Red Blood Cells for Diagnosing Glomerulonephritis

2019 ◽  
Vol 39 (3) ◽  
pp. 271
Author(s):  
Hyunjung Kim ◽  
Young Ok Kim ◽  
Yonggoo Kim ◽  
Jin-Soon Suh ◽  
Eun-Jung Cho ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Screening Tool ◽  
Cell Fraction

Leukoflow: Multi-Parameter Extended White Blood Cell Differentiation for Routine Analysis by Flow Cytometry

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4724-4724
Author(s):  
Gert-Jan M van de Geijn ◽  
Vincent van Rees ◽  
Natasja Bom ◽  
Hans Janssen ◽  
J.G. Pegels ◽  
...  
Keyword(s):  
Cell Surface ◽  
Blood Cell ◽  
White Blood Cell ◽  
Observer Variation ◽  
Cell Populations ◽  
Differential Count ◽  
Blood Samples ◽  
Cell Counter ◽  
Edta Blood ◽  
Myeloid Progenitors

Abstract Abstract 4724 Introduction Differential white blood cell count (dWBC) is an important and frequently used diagnostic tool in Hematology. Automated blood counters produce a five-part differential count. If the five-part differential count does not meet pre-set criteria, microscopic dWBC is performed. This morphological based dWBC is labour intensive and requires intensive and sustained training of technicians. In addition to inter-observer variation, the statistical variation is significant. Offering reliable round the clock service for dWBC can be a logistic challenge, in particular in samples from patients with haematological disease. Flowcytometry is a candidate reference method for dWBC. It has several advantages over morphological identification such as immunological definition of cell populations and high number of measured cells. Our goal was to develop a flowcytometric dWBC, called Leukoflow, which is easy to perform in a single tube, can be interpreted rapidly and can be available in a 24h/7d laboratory setting with a short turn around time. Method We selected 100 normal and 100 abnormal EDTA blood samples based on the data of the automated blood counter (LH750, Beckman Coulter) and the CLSI H20-A2 criteria. For flowcytometric dWBC, 20 ul EDTA blood is stained with an antibody cocktail (CD4, CD14, CD34, CD16, CD56, CD19, CD45, CD138, CD3 and CD71). Erythrocytes were lysed with ammonium chloride. Flowcount beads were added to determine the absolute concentrations of the cell populations in addition to their percentages. Flowcytometric analysis was performed using five channels on a FC500 (Beckman-Coulter). Using sequential gating, 13 cell populations were defined. For comparison, two independent technicians each counted 200 white blood cells. The data from Leukoflow are compared with the automated blood cell counter and the average from the two microscopical dWBCs. Results Leukoflow results correlate very well with both the automated blood cell counter and microscopic differentiation for leukocyte count as well as five-part differentiation. This applies for both normal and abnormal samples. Even without the use of positive markers for basophils or eosinophils, we could successfully define these populations by subtracting other positively defined populations in the regions where basophils and eosinophils are found in the CD45 SS staining. Reproducibility experiments showed that Leukoflow differentiation performed better than both traditional dWBC techniques. For all populations, except the myeloid progenitors, the coefficients of variation (CV%) of Leukoflow were less than 5%. Myeloid left-shift is detected earlier by Leukoflow in the abnormal samples. Furthermore blast counts reported by Leukoflow suffer less from inter-observer variation compared to manual dWBC, and proved to be more relevant and fitting to the clinical diagnosis. The correlation for erytroblasts between an additional flowcytometric CD45 and DRAQ5 based staining, and microscopy was excellent (r=0,96). In addition to traditional dWBC-techniques, extra cell populations are determined by Leukoflow: T-lymphocytes, CD4-lymphocytes, B-lymphocytes, NK cells, myeloid progenitors, plasma cells and blasts. When blasts are present, the Leukoflow analysis also indicates if they are from B-cell (surface CD19) or T-cell (surface CD3) origin. Conclusion Accurate dWBC can be performed with Leukoflow. The assay requires a small amount of blood and can be performed round the clock. The additional cell populations determined by Leukoflow enable faster diagnosis and give useful clinical information. The large number of cells analysed, compared with standard dWBC techniques, favors detection of rare cell populations. Preliminary data revealed that Leukoflow can also be used for analysis of bone marrow samples. Ongoing studies are focussing on the additional clinical value of Leukoflow over traditional dWBCs. Leukoflow is a highly interesting technique to screen blood samples from patients with haematological diseases in clinical haematology laboratories. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Detection of Transgenes in Gene Delivery Model Mice by Adenoviral Vector Using ddPCR

2019 ◽  
Author(s):  
Takehito Sugasawa ◽  
Kai Aoki ◽  
Koichi Watanabe ◽  
Koki Yanazawa ◽  
Tohru Natsume ◽  
...  
Keyword(s):  
Blood Cell ◽  
Quantitative Pcr ◽  
Whole Blood ◽  
Digital Pcr ◽  
Cell Fraction ◽  
Detection Methods ◽  
Gene Doping ◽  
World Anti Doping Agency ◽  
Injection Methods ◽  
Tail Tip

With the rapid progress of genetic engineering and gene therapy, World Anti-Doping Agency has alerted to gene doping and prohibited its use in sports. However, there is no standard method available yet for detection of transgenes delivered by recombinant adenoviral (rAdV) vectors. Here we aimed to develop a detection method for transgenes delivered by rAdV vectors in a mouse model that mimics gene doping. rAdV vectors containing mCherry gene was delivered in mice through intravenous injection or local muscular injection. After five days, stool and whole blood samples were collected, and total DNA was extracted. As additional experiments, whole blood was also collected from mouse tail tip until 15 days from injection of the rAdv vector. Transgene fragments from different DNA samples were analyzed using semi-quantitative PCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene fragments could directly be detected from blood cell fraction-DNA, plasma-cell free DNA and stool-DNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a combination of blood cell fraction-DNA and ddPCR was more sensitive than other combinations used in this model. These results could accelerate the development of detection methods for gene doping.

scholarly journals Small Red Blood Cell Fraction on the UF-1000i Urine Analyzer as a Screening Tool to Detect Dysmorphic Red Blood Cells for Diagnosing Glomerulonephritis

2019 ◽  
Vol 39 (3) ◽  
pp. 271-277 ◽  
Author(s):  
Hyunjung Kim ◽  
Young Ok Kim ◽  
Yonggoo Kim ◽  
Jin-Soon Suh ◽  
Eun-Jung Cho ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Screening Tool ◽  
Cell Fraction

scholarly journals Detection of Transgenes in Gene Delivery Model Mice by Adenoviral Vector Using ddPCR

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 436 ◽  
Author(s):  
Takehito Sugasawa ◽  
Kai Aoki ◽  
Koichi Watanabe ◽  
Koki Yanazawa ◽  
Tohru Natsume ◽  
...  
Keyword(s):  
Blood Cell ◽  
Quantitative Pcr ◽  
Whole Blood ◽  
Digital Pcr ◽  
Cell Fraction ◽  
Detection Methods ◽  
Gene Doping ◽  
World Anti Doping Agency ◽  
Injection Methods ◽  
Tail Tip

With the rapid progress of genetic engineering and gene therapy, the World Anti-Doping Agency has been alerted to gene doping and prohibited its use in sports. However, there is no standard method available yet for the detection of transgenes delivered by recombinant adenoviral (rAdV) vectors. Here, we aim to develop a detection method for transgenes delivered by rAdV vectors in a mouse model that mimics gene doping. These rAdV vectors containing the mCherry gene was delivered in mice through intravenous injection or local muscular injection. After five days, stool and whole blood samples were collected, and total DNA was extracted. As additional experiments, whole blood was also collected from the mouse tail tip until 15 days from injection of the rAdv vector. Transgene fragments from different DNA samples were analyzed using semi-quantitative PCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene fragments could be directly detected from blood cell fraction DNA, plasma cell-free DNA, and stool DNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a combination of blood cell fraction DNA and ddPCR was more sensitive than other combinations used in this model. These results could accelerate the development of detection methods for gene doping.

Reduction of Prion Infectivity in Packed Red Cells by Separation of Whole Blood and Washing of the Red Cell Fraction.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2890-2890
Author(s):  
Rodrigo Morales ◽  
Kimberley A. Buytaert-Hoefen ◽  
Eric T. Hansen ◽  
Dennis Hlavinka ◽  
Raymond Goodrich ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Red Cells ◽  
Cell Fraction ◽  
Health Concern ◽  
Red Cell ◽  
New Variant

Abstract Although prion diseases are rare in humans, the established link between a new variant form of CJD (vCJD) and the consumption of cattle meat contaminated by BSE have raised concerns about a possible outbreak of a large epidemic in the human population. Over the past few years, BSE has become a significant health concern in several countries, and it now seems apparent that vCJD can also be iatrogenically transmitted from human to human by blood transfusion. Exacerbating this state of affairs is the lack of a reliable test to identify individuals incubating the disease during the long and silent period from the onset of infection to the appearance of clinical symptoms. The purpose of this research study was to evaluate the effectiveness of separation of whole blood and washing of the red cell fraction for the removal of infectious scrapie prion protein (PrPSc) from red blood cell (RBC) suspensions. Samples of human, whole blood were spiked with 5 × 106 LD50 263K PrPSc. Analysis of the treated sample supernatants by Western blot revealed that approximately >88% of the PrPSc was removed with the initial plasma expression and the equivalent of 6% was detected in a saline wash (300 mL; 0.9% saline). The final sample of RBCs revealed no detectable levels of PrPSc by Western blots. Further analysis of the treated RBCs using the PMCA assay indicated detectable amounts of PrPSc only after 2 consecutive amplification rounds. Semi-quantitative analysis of PMCA amplification enabled us to estimate that the treated RBCs contained less than 1 × 104 LD50 PrPSc. This corresponded to removal levels exceeding ≥99% of spiked material in whole blood. These in vitro estimations were confirmed by in vivo infectivity studies in a hamster model of disease transmission. Results from in vivo studies displayed significant differences in the incubation periods of the spiked blood inoculated hamsters (100.1 ± 1.7) versus washed RBCs (135.8 ± 6.7). Moreover, a substantial difference in the attack rate (6/15: 40% in washed RBC, versus 13/13: 100% in spiked blood) further indicated a substantial removal of infectious prions. Comparison of this data with results of animals inoculated with different dilutions of infectious material, indicated a >99.94% reduction of infectivity. Washed, packed human red cells produced by this procedure were able to be stored in standard additive solutions (AS-3) for 42 days while still meeting all in vitro blood bank standards for acceptable red cell quality. Conclusion This data suggests that separation of plasma coupled with a simple, low volume wash of red cells may represent an efficient method to remove prions from red blood cell fractions, thus reducing possible infectivity of these products.

Applications of deep learning to the assessment of red blood cell deformability

Biorheology ◽  
2021 ◽  
pp. 1-10
Author(s):  
Alper Turgut ◽  
Özlem Yalçin
Keyword(s):  
Neural Network ◽  
Theoretical Model ◽  
Blood Cell ◽  
Convolutional Neural Network ◽  
Red Blood Cell ◽  
Network Architecture ◽  
Laser Diffraction ◽  
Cell Fraction ◽  
Cell Deformability ◽  
Diffraction Patterns

BACKGROUND: Measurement of abnormal Red Blood Cell (RBC) deformability is a main indicator of Sickle Cell Anemia (SCA) and requires standardized quantification methods. Ektacytometry is commonly used to estimate the fraction of Sickled Cells (SCs) by measuring the deformability of RBCs from laser diffraction patterns under varying shear stress. In addition to estimations from model comparisons, use of maximum Elongation Index differences (ΔEImax) at different laser intensity levels was recently proposed for the estimation of SC fractions. OBJECTIVE: Implement a convolutional neural network to accurately estimate rigid-cell fraction and RBC concentration from laser diffraction patterns without using a theoretical model and eliminating the ektacytometer dependency for deformability measurements. METHODS: RBCs were collected from control patients. Rigid-cell fraction experiments were performed using varying concentrations of glutaraldehyde. Serial dilutions were used for varying the concentration of RBC. A convolutional neural network was constructed using Python and TensorFlow. RESULTS: Our measurements and model predictions show that a linear relationship between ΔEImax and rigid-cell fraction exists only for rigid-cell fractions less than 0.2. Our proposed neural network architecture can be used successfully for both RBC concentration and rigid-cell fraction estimations without a need for a theoretical model.

IMPDH activity in whole blood and isolated blood cell fraction for monitoring of cellcept-mediated immunosuppression

1999 ◽  
Vol 31 (1-2) ◽  
pp. 1115-1116 ◽  
Author(s):  
M Storck ◽  
D Abendroth ◽  
W Albrecht ◽  
H.W Sollinger
Keyword(s):  
Blood Cell ◽  
Whole Blood ◽  
Cell Fraction ◽  
Impdh Activity
Sign in / Sign up

Export Citation Format

Share Document