How I investigate dysgranulopoiesis

Shekhar, Roshini; Srinivasan, Vishrut K.; Pai, Swati

doi:10.1111/ijlh.13607

Cited by 8 publications

(7 citation statements)

References 62 publications

(66 reference statements)

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“…The most common method for routine diagnosis of hematological pathologies is complete blood count (CBC), which measures cell volume based on a combination of electrical impedance and scatter analysis, as well as non-specific fluorescence labels to provide automated, low-cost, fast, and fairly reliable estimated five-part differential counting of WBCs: neutrophils, monocytes, eosinophils, basophils, and lymphocytes. However, for the more detailed morphological analyses of WBCs needed to diagnose MDS, a blood smear is needed, as this test can detect morphological abnormalities such as nuclear hyper-segmentation and cytoplasmic hypo-granularity [ 7 ]. In a blood smear, typically, manual analysis is performed under a light microscope, which is a labor-intensive method requiring skilled technicians and can take a long time [ 23 ].…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, infections are a major cause of illness and death in MDS patients [ 5 ], both in patients with neutrophil dysfunction and in patients with neutropenia [ 6 ]. Morphological and physiological abnormalities are common in neutrophils of MDS patients, including nuclear hyper-segmentation, cytoplasmic hypo-granularity [ 7 ], and a reduction in reactive oxygen species (ROS), which are essential for the management of pathogens [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

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Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Barnea,

Luria,

Girsault

et al. 2024

Bioengineering

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Myelodysplastic syndromes (MDSs) are a group of potentially deadly diseases that affect the morphology and function of neutrophils. Rapid diagnosis of MDS is crucial for the initiation of treatment that can vastly improve disease outcome. In this work, we present a new approach for detecting morphological differences between neutrophils isolated from blood samples of high-risk MDS patients and blood bank donors (BBDs). Using fluorescent flow cytometry, neutrophils were stained with 2′,7′-dichlorofluorescin diacetate (DCF), which reacts with reactive oxygen species (ROS), and Hoechst, which binds to DNA. We observed that BBDs possessed two cell clusters (designated H and L), whereas MDS patients possessed a single cluster (L). Later, we used FACS to sort the H and the L cells and used interferometric phase microscopy (IPM) to image the cells without utilizing cell staining. IPM images showed that H cells are characterized by low optical path delay (OPD) in the nucleus relative to the cytoplasm, especially in cell vesicles containing ROS, whereas L cells are characterized by low OPD in the cytoplasm relative to the nucleus and no ROS-containing vesicles. Moreover, L cells present a higher average OPD and dry mass compared to H cells. When examining neutrophils from MDS patients and BBDs by IPM during flow, we identified ~20% of cells as H cells in BBDs in contrast to ~4% in MDS patients. These results indicate that IPM can be utilized for the diagnosis of complex hematological pathologies such as MDS.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Barnea,

Luria,

Girsault

et al. 2024

Bioengineering

View full text Add to dashboard Cite

show abstract

“…The most common method for routine diagnosis of hematological pathologies is complete blood count (CBC), which measures cell volume based on a combination of electrical impedance and scatter analysis, as well as non-specific fluorescence labels to provide automated, low-cost, fast, and fairly reliable estimated five-part differential counting of WBCs: neutrophils, monocytes, eosinophils, basophils, and lymphocytes. However, for the more detailed morphological analyses of WBCs needed to diagnose MDS, a blood smear is needed, as this test can detect morphological abnormalities such as nuclear hypersegmentation and cytoplasmic hypo-granularity [7]. In a blood smear, typically, manual analysis is performed under a light microscope, which is a labor-intensive method requiring skilled technicians and can take a long time [23].…”

Section: Discussionmentioning

confidence: 99%

Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Barnea,

Luria,

Girsault

et al. 2024

Preprint

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(1) Background: Myelodysplastic syndromes (MDS) are a group of diseases that affect the morphology and function of neutrophils. Rapid diagnosis of MDS is crucial for the initiation of treatment that can vastly improve disease outcome. (2) methods: In this work, we present a new approach for detecting morphological differences between neutrophils isolated from blood samples of high risk MDS patients and that for patients and blood bank donors (BBDs). First, using fluorescent flow cytometry CD66b positive cells were stained with 2′,7′-dichlorofluorescin diacetate (DCF) and Hoechst. Secondly, we imaged the cells using interferometric phase microscopy (IPM) without utilizing cell staining. (3) Results: flow cytometry results show MDS patients present different dispersion patterns compared to BBDs. BBDs present two distinct subpopulations (one of these are the H cells constituting 16% for neutrophils) whereas MDS patients present only 4% of this subpopulation. IPM results are consistent with this pattern: ~20% of BBDs neutrophils were characterized as H cells and whereas MDS patients present only ~4% of these cells. (5) Conclusion- IPM can be utilized for diagnosis of complex hematological pathologies such as MDS.

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“…However, dysgranulopoiesis is not unique to MDS. Once dysgranulopoiesis is identified according to the abnormal NeuX and NeuZ value as well as microscope observation, correlation with CBC, underlying medical conditions, biochemical and other ancillary investigations is necessary to rule out non‐neoplastic causes 8 . Therefore, NeuX and NeuZ can be used to predict neutrophil dysplasia, but do not directly point to a diagnosis of MDS.…”

Section: Discussionmentioning

confidence: 99%

“…6,7 Recent studies have found that CPD parameters generated by automatic hematology analyzer, reflecting leukocyte cellular characteristics like cell complexity or granularity, are possible to use to detect dysgranulopoiesis. 8 Since then, researchers have carried out a large number of studies using different brands of automatic hematology analyzer, all suggesting that there are differences in hematological parameters of peripheral blood samples of healthy people and MDS patients. [9][10][11][12][13] Various studies have found that neutrophil parameter NeuX provided by the automatic hematology analyzer was correlated with the morphological change of neutrophil decreased granules or agranularity.…”

Section: Discussionmentioning

confidence: 99%

Combination of NeuX and NeuZ can predict neutrophil dysplasia features of myelodysplastic neoplasms in peripheral blood

Sun

Zhang

et al. 2023

Int J Lab Hematology

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Introduction:The assessment of neutrophil dysplasia features in peripheral blood is very helpful for the early screening and diagnosis of myelodysplastic neoplasms (MDS). Cell population data (CPD) parameters generated by automated hematology analyzers can reflect morphological characteristics of blood cells. This study aimed to investigate the clinical significance of CPD parameters neutrophil (Neu) X, NeuY and NeuZ in assessing neutrophil dysplasia.Methods: 218 MDS patients were divided into two subgroups according to neutrophil morphology. The differences of neutrophil research parameters between the two MDS subgroups and the control group, consisting of 210 healthy individuals, were compared, the correlation among neutrophil research parameters and the relationship between these parameters and cell morphology in MDS patients were analyzed, and receiver operating characteristic analysis were performed. Results: The median values of neutrophil research parameters NeuX and NeuZ in MDSwith granulocyte dysplasia group were significantly lower than those in MDS without granulocyte dysplasia group and control group (p < 0.001), and they were positively correlated (r = 0.878, p < 0.001). The area under the receiver operating characteristic curve of NeuX and NeuZ was 0.720 (95% CI: 0.643-0.796, p < 0.001) and 0.738 (95% CI: 0.665-0.811, p < 0.001), respectively. In addition, with the decrease of NeuX value, neutrophils gradually show decreased nuclear segment and/or cytoplasmic granules.Conclusions: Combining NeuX and NeuZ can predict neutrophil dysplasia features of MDS in peripheral blood, and this can be an easier method to screen for the neutrophil dysplasia cases, as compared with the microscopic examination of peripheral blood and/or bone marrow smears.

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How I investigate dysgranulopoiesis

Cited by 8 publications

References 62 publications

Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry

Combination of NeuX and NeuZ can predict neutrophil dysplasia features of myelodysplastic neoplasms in peripheral blood

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