Diagnostic utility of flow cytometric immunophenotyping in cerebrospinal fluid specimen

2021 ◽  
Vol 156 (Supplement_1) ◽  
pp. S121-S121
Author(s):  
N Naiyer ◽  
H Lin
Keyword(s):  
Flow Cytometry ◽  
Hematologic Malignancy ◽  
Flow Cytometric Analysis ◽  
Final Diagnosis ◽  
Hematologic Malignancies ◽  
Prior History ◽  
Flow Cytometric Immunophenotyping ◽  
Flow Cytometric ◽  
Review Period ◽  
History Of

Abstract Introduction/Objective Flow cytometric immunophenotyping (FCI) of cerebrospinal fluids (CSF) has increasingly been used for diagnosing and monitoring hematologic malignancies. We reviewed CSF specimens sent for flow cytometry evaluation to identify the utility and limitations of FCI. Methods/Case Report We performed a retrospective review on CSF specimens received from July to October 2020. Samples were sent with requisition for unexplained neurologic signs or symptoms, to determine CNS involvement of neoplasm, or to stage a neoplasm. We reviewed specimen volume and cell count of flow cytometry samples and electronic medical records for possible diagnosis at time of specimen submission, final diagnosis, and concurrent cytology diagnosis. Results (if a Case Study enter NA) A total of 104 CSF samples from 59 patients were processed by the Flow Cytometry Laboratory during the review period. Of the 104 samples, 66% were from patients with a prior history of a hematologic malignancy, of which 20% had abnormal findings by FCI and cytomorphology. FCI was noncontributory for the cases in which patient did not have a previously diagnosed hematologic malignancy and underwent lumbar puncture for neurological abnormalities (n = 34). Concurrent cytology results were available in all but one case. Atypical or reactive findings by cytomorphology were seen in 21 cases (20%), for which flow cytometry studies showed no diagnostic immunophenotype in 12 cases. Abnormal FCI results occurred in 5 cases with history of hematologic malignancies, while concurrent cytomorphologic reports were negative. Conclusion Our findings suggest that CSF flow cytometry has low utility for screening patients with undifferentiated neurologic symptoms or without a prior hematologic malignancy history. Use as a screening tool for cases without clinical suspicion of hematolymphoid neoplasm is debatable. Flow cytometric and cytomorphologic analysis should be performed concurrently. While flow cytometric analysis in CSF may have higher sensitivity in hematologic malignancies, cytomorphology appears more favorable in identifying atypia or reactive conditions.

Flow Cytometric Immunophenotyping of Bone Marrow Core Biopsies

1999 ◽  
Vol 123 (3) ◽  
pp. 206-212 ◽  
Author(s):  
Cherie H. Dunphy ◽  
Frank R. Dunphy ◽  
John L. Visconti
Keyword(s):  
Bone Marrow ◽  
Core Biopsy ◽  
Hematologic Malignancy ◽  
Data Extraction ◽  
Final Diagnosis ◽  
Hematologic Malignancies ◽  
Cell Of Origin ◽  
Flow Cytometric Immunophenotyping ◽  
Flow Cytometric ◽  
Biopsy Specimens

Abstract Objective.—To report a method for flow cytometric immunophenotyping (FCI) bone marrow (BM) core biopsies in patients with hematologic malignancies of the BM who present with a failed BM aspiration (“dry tap”). Design and Setting.—Core biopsy specimens of BM were obtained from 8 patients who presented with previously undiagnosed hematologic malignancies arising in (7 cases) or secondarily involving (1 case) the BM and a dry tap. Suspensions of the BM core biopsy specimens were prepared and analyzed by FCI methods. Data Extraction and Data Synthesis.—The FCI data were analyzed in conjunction with cytomorphologic, histologic, immunohistochemical, and cytogenetic findings in all cases to determine a final diagnosis. Conclusions.—The prepared BM core suspensions were viable and allowed for a complete immunophenotype profile by FCI in all cases, resulting in a clear definition of the cell of origin of the hematologic malignancy. Because of lack of preservation of architectural features and the potential for artifactual alterations of the relative frequency of abnormal cells, the FCI data must always be correlated with histologic sections of the BM biopsy.

scholarly journals Flow Cytometric Analysis of Cerebrospinal Fluid Has Low Diagnostic Yield in Samples Without Atypical Morphology or Prior History of Hematologic Malignancy

2014 ◽  
Vol 141 (4) ◽  
pp. 515-521 ◽  
Author(s):  
Angela M. B. Collie ◽  
Brian T. Hill ◽  
Glen H. J. Stevens ◽  
Kathleen Fenner ◽  
Elizabeth Gazdick ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Diagnostic Yield ◽  
Hematologic Malignancy ◽  
Flow Cytometric Analysis ◽  
Prior History ◽  
Flow Cytometric ◽  
History Of

Diagnostic utility of flow cytometric immunophenotyping in myelodysplastic syndrome

Blood ◽  
2001 ◽  
Vol 98 (4) ◽  
pp. 979-987 ◽  
Author(s):  
Maryalice Stetler-Stevenson ◽  
Diane C. Arthur ◽  
Nicholas Jabbour ◽  
Xiu Y. Xie ◽  
Jeff Molldrem ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Diagnostic Utility ◽  
Flow Cytometric Immunophenotyping ◽  
Flow Cytometric ◽  
Healthy Donors ◽  
Number Of Patients ◽  
Highly Sensitive ◽  
History Of ◽  
Diagnosis And Classification

The myelodysplastic syndromes (MDSs) are characterized by bilineage or trilineage dysplasia. Although diagnostic criteria are well established for MDS, a significant number of patients have blood and bone marrow findings that make diagnosis and classification difficult. Flow cytometric immunophenotyping is an accurate and highly sensitive method for detection of quantitative and qualitative abnormalities in hematopoietic cells. Flow cytometry was used to study hematopoietic cell populations in the bone marrow of 45 patients with straightforward MDS. The results were compared with those obtained in a series of patients with aplastic anemia, healthy donors, and patients with a history of nonmyeloid neoplasia in complete remission. The immunophenotypic abnormalities associated with MDS were defined, and the diagnostic utility of flow cytometry was compared, with morphologic and cytogenetic evaluations in 20 difficult cases. Although morphology and cytogenetics were adequate for diagnosis in most cases, flow cytometry could detect immunophenotypic abnormalities in cases when combined morphology and cytogenetics were nondiagnostic. It is concluded that flow cytometric immunophenotyping may help establish the diagnosis of MDS, especially when morphology and cytogenetics are indeterminate.

scholarly journals Flow Cytometric, Morphologic, and Laboratory Comparative Study in Patients With Leukocytosis and Cytopenia

2019 ◽  
Vol 153 (2) ◽  
pp. 266-273
Author(s):  
Estafani Rivas ◽  
Fred V Plapp ◽  
Wei Cui
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood ◽  
Screening Test ◽  
Hematologic Malignancy ◽  
Cost Effective ◽  
Effective Screening ◽  
Flow Cytometric ◽  
Blood Smears ◽  
History Of ◽  
Peripheral Blood Smears

Abstract Objectives We wanted to evaluate the effectiveness of flow cytometry immunophenotyping (FCI) as a screening test for patients with leukocytosis and cytopenia. Methods We identified 320 patients during August 2016 to December 2016 and evaluated FCI and morphology of peripheral blood smears (PBSs). Results The most common indications for FCI included history of hematologic malignancy (HHM, n = 126), leukocytosis (n = 80), and cytopenia (n = 53). Positive FCI rate was low with a range of 4.4% to 12.5% in patients with absolute neutrophilia regardless of HHM, if cases with circulating blasts were excluded. Patients with absolute lymphocytosis had a 93% positive FCI rate. Patients with HHM and pancytopenia showed a higher incidence of positive FCI findings than patients without HHM and with isolated cytopenia. PBS morphology correlated strongly with FCI (P = .0001). Conclusion PBS evaluation is an accurate and cost-effective screening test. FCI for patients with mature neutrophilia and isolated cytopenia has a very low yield.

scholarly journals Multiparameter Flow Cytometric Analysis of Antibiotic Effects on Membrane Potential, Membrane Permeability, and Bacterial Counts of Staphylococcus aureus andMicrococcus luteus

2000 ◽  
Vol 44 (4) ◽  
pp. 827-834 ◽  
Author(s):  
David J. Novo ◽  
Nancy G. Perlmutter ◽  
Richard H. Hunt ◽  
Howard M. Shapiro
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Membrane Potential ◽  
Membrane Permeability ◽  
Micrococcus Luteus ◽  
Flow Cytometric Analysis ◽  
Bacterial Counts ◽  
Flow Cytometric ◽  
Permeability Parameter ◽  
Particle Counts

ABSTRACT Although flow cytometry has been used to study antibiotic effects on bacterial membrane potential (MP) and membrane permeability, flow cytometric results are not always well correlated to changes in bacterial counts. Using new, precise techniques, we simultaneously measured MP, membrane permeability, and particle counts of antibiotic-treated and untreated Staphylococcus aureus andMicrococcus luteus cells. MP was calculated from the ratio of red and green fluorescence of diethyloxacarbocyanine [DiOC2(3)]. A normalized permeability parameter was calculated from the ratio of far red fluorescence of the nucleic acid dye TO-PRO-3 and green DiOC2(3) fluorescence. Bacterial counts were calculated by the addition of polystyrene beads to the sample at a known concentration. Amoxicillin increased permeability within 45 min. At concentrations of <1 μg/ml, some organisms showed increased permeability but normal MP; this population disappeared after 4 h, while bacterial counts increased. At amoxicillin concentrations above 1 μg/ml, MP decreased irreversibly and the particle counts did not increase. Tetracycline and erythromycin caused smaller, dose- and time-dependent decreases in MP. Tetracycline concentrations of <1 μg/ml did not change permeability, while a tetracycline concentration of 4 μg/ml permeabilized 50% of the bacteria; 4 μg of erythromycin per ml permeabilized 20% of the bacteria. Streptomycin decreased MP substantially, with no effect on permeability; chloramphenicol did not change either permeability or MP. Erythromycin pretreatment of bacteria prevented streptomycin and amoxicillin effects. Flow cytometry provides a sensitive means of monitoring the dynamic cellular events that occur in bacteria exposed to antibacterial agents; however, it is probably simplistic to expect that changes in a single cellular parameter will suffice to determine the sensitivities of all species to all drugs.

scholarly journals Chromosome doubling in Cattleya tigrina A. Rich

2019 ◽  
Vol 15 (11) ◽  
Author(s):  
Thays Saynara Alves Menezes-Sá ◽  
Maria de Fátima Arrigoni-Blank ◽  
Andréa Santos da Costa ◽  
Janay De Almeida Santos-Serejo ◽  
Arie Fitzgerald Blank ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Propidium Iodide ◽  
Chromosome Doubling ◽  
Flow Cytometric Analysis ◽  
Leaf Tissue ◽  
Stomatal Index ◽  
Chromosome Duplication ◽  
Flow Cytometric ◽  
Young Leaves ◽  
Commercial Value

Chromosome doubling induction in orchids may benefit their production for resulting in flowers of higher commercial value, larger size and higher content of substances that intensify the color and fragrance when compared with diploid orchids. This work aimed to induce and confirm artificial polyploidization, using flow cytometry and stomatal analysis. Explants were treated with colchicine at concentrations of 0, 2.5, 7.5, and 12.5 mM, for 24 and 48 hours and with oryzalin, at concentrations of 0, 10, 30, and 50 μM, for three and six days. For the flow cytometric analysis, a sample of leaf tissue was removed from each plant, crushed to release the nuclei and stained with propidium iodide. In addition to flow cytometry, the ploidy of the antimitotic treated plants was evaluated by stomata analysis. Young leaves were used where the density, functionality and stomatal index were evaluated. Colchicine provided induction of satisfactory polyploidy in C. tigrina at all concentrations and times of exposure, obtaining a greater number of polyploid individuals in the concentration of 12.5 mM for 48 hours. Oryzalin did not induce chromosome duplication at the tested concentrations.

The Value of CD64 Expression in Distinguishing Acute Myeloid Leukemia With Monocytic Differentiation From Other Subtypes of Acute Myeloid Leukemia: A Flow Cytometric Analysis of 64 Cases

2007 ◽  
Vol 131 (5) ◽  
pp. 748-754
Author(s):  
Cherie H. Dunphy ◽  
Wohzan Tang
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Flow Cytometric Analysis ◽  
World Health ◽  
Monocytic Differentiation ◽  
Flow Cytometric Immunophenotyping ◽  
Cd64 Expression ◽  
Flow Cytometric ◽  
Health Organization ◽  
Acute Myeloid

Abstract Context.—Flow cytometric immunophenotyping is a useful ancillary tool in the diagnosis and subclassification of acute myeloid leukemias (AMLs). A recent study concluded that CD64 is sensitive and specific for distinguishing AMLs with a monocytic component (ie, AML M4 and AML M5) from other AML subtypes. However, in that study, the intensity of CD64 was not well defined and the number of non-M4/non-M5 AMLs was small. Objective.—To evaluate the usefulness of CD64 by flow cytometric immunophenotyping in distinguishing AMLs with monocytic differentiation from other AML subtypes. Design.—Sixty-four AMLs subclassified based on the French-American-British and World Health Organization classifications on pretreatment bone marrows were retrieved from our files (7 M0s, 11 M1s, 17 M2s, 7 M3s, 9 M4s, 7 M5s, 4 M6s, and 2 M7s). A standard panel of markers, including CD2, CD3, CD5, CD7, CD10, CD11b, CD13, CD14, CD15, CD19, CD20, CD33, CD34, CD45, CD56, CD64, CD117, and HLA-DR, were analyzed by flow cytometric immunophenotyping in all AMLs (52 bone marrow samples; 12 peripheral blood samples). Results.—CD64 was expressed in AML subtypes M0 to M5 in varying intensities: heterogeneously expressed in 1 of 7 M0s; dimly expressed in 3 of 11 M1s; dimly and moderately expressed in 6 and 2 of 17 M2s, respectively; dimly and moderately expressed in 5 and 1 of 7 M3s, respectively; dimly expressed in 4 of 9 M4s; and heterogeneously, moderately, and strongly expressed in 1, 3, and 3 of 7 M5s, respectively. Conclusions.—Strong CD64 expression distinguishes AML M5; however, heterogeneous, dim, or moderate expression in itself does not distinguish M0 through M4 subtypes from M5 with dim to moderate CD64 expression. However, any CD64 expression associated with strong CD15 expression distinguishes AML M4 or M5, from other AML subtypes.

scholarly journals Flow cytometric analysis of herpes simplex virus type 1 susceptibility to acyclovir, ganciclovir, and foscarnet.

1997 ◽  
Vol 41 (12) ◽  
pp. 2686-2692 ◽  
Author(s):  
I Pavić ◽  
A Hartmann ◽  
A Zimmermann ◽  
D Michel ◽  
W Hampl ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Cytopathic Effect ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Resistant Strains ◽  
Simplex Virus

We established a quantitative flow cytometric method for determination of herpes simplex virus type 1 (HSV-1) susceptibility to acyclovir (ACV), ganciclovir, and foscarnet in vitro. Susceptibility was defined in terms of the drug concentration which reduced the number of cells expressing HSV-1 glycoprotein C (gpC) with a fluorescence intensity of > or =10(2) by 50% (IC50). Flow cytometry allowed us to use a high (1.0) as well as a low (0.005) multiplicity of infection, and determination of the IC50 was possible after one or more viral replicative cycles. IC50s were dependent on virus input and on time postinfection. In mixture experiments, 1 to 2% resistant viruses added to a sensitive strain could be detected. The results obtained by flow cytometry showed a good qualitative correlation with those achieved by cytopathic effect inhibitory assay. However, flow cytometry might detect more quantitative differences in drug susceptibility, especially among resistant strains, as confirmed also by determination of intracellular drug phosphorylation. The mean IC50s for ACV-sensitive strains were 0.45 to 1.47 microM, and those for ACV-resistant strains were between 140 and 3,134 microM. Flow cytometric analysis was fast and accurate, automatizable, and highly reproducible. Flow cytometry may be a more powerful tool than standard cytopathic effect-based assays and could have advantages for the detection of low levels of drug resistance or mixtures of sensitive and resistant virus strains.

scholarly journals Flow cytometric analysis of virus-like particles and heterotrophic bacteria within coral-associated reef water

2006 ◽  
Vol 86 (3) ◽  
pp. 563-566 ◽  
Author(s):  
Nicole L. Patten ◽  
Justin R. Seymour ◽  
James G. Mitchell
Keyword(s):  
Flow Cytometry ◽  
Bacterial Community ◽  
Heterotrophic Bacteria ◽  
Flow Cytometric Analysis ◽  
Water Layer ◽  
Dead Coral ◽  
Overlying Water ◽  
Virus Like Particles ◽  
Flow Cytometric ◽  
Diseased Coral

Using flow cytometry, two distinct populations of virus-like particles (VLP) and heterotrophic bacteria were defined within the 12 cm water layer immediately overlying healthy, diseased and dead acroporid corals. Bacterial abundances were similar in overlying water for all coral types, however, VLP were 30% higher above diseased corals than healthy or dead corals. Mean virus to bacteria ratios (VBR) were up to 30% higher above diseased corals than above healthy or dead coral or in distant water. Concomitant with increasing VLP concentrations within 5 cm of coral surfaces, VBR distributions were generally highest above healthy and diseased coral and depressed above dead coral. These results suggest fundamental shifts in the VLP and bacterial community in water associated with diseased corals.

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