Elimination of light scatter interference in dual-laser flow cytometry by synchronous detection of emitted fluorescence: theory and demonstration using simulated signals

1999 ◽  
Author(s):  
John A. Steinkamp ◽  
Jan F. Keij
Keyword(s):  
Flow Cytometry ◽  
Light Scatter ◽  
Synchronous Detection ◽  
Dual Laser

scholarly journals Identification and Phenotyping of Leukocytes in Bovine Bronchoalveolar Lavage Fluid

2004 ◽  
Vol 11 (4) ◽  
pp. 795-798 ◽  
Author(s):  
E. C. Soethout ◽  
K. E. Müller ◽  
A. J. M. van den Belt ◽  
V. P. M. G. Rutten
Keyword(s):  
Flow Cytometry ◽  
Bronchoalveolar Lavage ◽  
Alveolar Macrophages ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Light Scatter ◽  
Specific Antibodies ◽  
Dual Laser

ABSTRACT A method is proposed to identify leukocyte subpopulations in bovine bronchoalveolar lavage fluid by dual-laser flow cytometry. The technique uses several parameters, i.e., exclusion of highly autofluorescent alveolar macrophages and inclusion of leukocytes on the basis of labeling by specific antibodies and light scatter characteristics.

scholarly journals Detection and analysis by dual-laser flow cytometry of bacteriophage T4 DNA insideEscherichia coli

Cytometry ◽  
1991 ◽  
Vol 12 (2) ◽  
pp. 167-171 ◽  
Author(s):  
Cynthia A. Sanders ◽  
David M. Yajko ◽  
Patricia S. Nassos ◽  
William C. Hyun ◽  
Mack J. Fulwyler ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bacteriophage T4 ◽  
Dual Laser

scholarly journals A Multiplexed Assay That Monitors Effects of Multiple Compound Treatment Times Reveals Candidate Immune-Enhancing Compounds

2018 ◽  
Vol 23 (7) ◽  
pp. 646-655 ◽  
Author(s):  
Ziyan Zhao ◽  
Liza Henowitz ◽  
Adam Zweifach
Keyword(s):  
Flow Cytometry ◽  
Cytotoxic T Lymphocytes ◽  
High Throughput ◽  
Treatment Time ◽  
Single Sample ◽  
Light Scatter ◽  
Flow Cytometry Assay ◽  
Multiple Treatment ◽  
Lytic Granule ◽  
Activation Status

We previously developed a flow cytometry assay that monitored lytic granule exocytosis in cytotoxic T lymphocytes stimulated by contacting beads coated with activating anti-CD3 antibodies. That assay was multiplexed in that responses of cells that did or did not receive the activating stimulus were distinguished via changes in light scatter accompanying binding of cells to beads, allowing us to discriminate compounds that activate responses on their own from compounds that enhance responses in cells that received the activating stimulus, all within a single sample. Here we add a second dimension of multiplexing by developing means to assess in a single sample the effects of treating cells with test compounds for different times. Bar-coding cells before adding them to test wells lets us determine compound treatment time while also monitoring activation status and response amplitude at the point of interrogation. This multiplexed assay is suitable for screening 96-well plates. We used it to screen compounds from the National Cancer Institute, identifying several compounds that enhance anti-LAMP1 responses. Multiple-treatment-time (MTT) screening enabled by bar-coding and read via high-throughput flow cytometry may be a generally useful method for facilitating the discovery of compounds of interest.

Adrenalectomy induces a decrease in the light scatter properties and amylase content of isolated zymogen granules from rat pancreas as analyzed by flow cytometry

1995 ◽  
Vol 147 (3) ◽  
pp. 431-440 ◽  
Author(s):  
A C Garcia-Montero ◽  
I De Dios ◽  
A I Rodriguez ◽  
A Orfao ◽  
M A Manso
Keyword(s):  
Flow Cytometry ◽  
Progressive Increase ◽  
Enzyme Concentration ◽  
Male Rats ◽  
Zymogen Granule ◽  
Light Scatter ◽  
Zymogen Granules ◽  
Mean Size ◽  
A Minor ◽  
Adrenalectomized Rats

Abstract The effect of glucocorticoid deprivation induced in male rats by adrenalectomy on the pancreatic zymogen granules was studied. Zymogen granules were purified from control, sham-operated and adrenalectomized animals studied 1, 3 and 7 days after surgery. The zymogen granules were characterized by flow cytometry, and in each granule the size (based on the forward or low angle light scatter (FSC) parameter), membrane complexity (based on side or 90° light scatter (SSC) parameter) and amylase content were evaluated. Amylase content/DNA ratio in pancreatic homogenates was also analyzed. The zymogen granules of the control rats were found to be distributed in two populations: a major one – R1 (95·45 ± 1·21%) – containing zymogen granules with a smaller mean size and complexity, and a minor population - R2 (4·45 ± 0·24%) – the granules of which had a mean size which was larger and more complex. At day +1 after adrenalectomy the zymogen granules were significantly (P<0·05) smaller than those of control animals. The R2 zymogen granules were similar to those from R1 as regards their size, but were more complex, suggesting that the immediate effect of glucocorticoid deprivation is to induce a depletion of the larger granules presumably belonging to the R2 population. The amount of amylase per granule did not vary at day +1 after adrenalectomy, although the amylase content/size ratio per granule was significantly (P<0·001) increased. This mechanism could be explained in terms of the existence of a bypass defined in the adrenalectomized animals between the granular content and cytosolic enzymes. Prolongation of the adrenalectomy period to 3 and 7 days resulted in a progressive increase in zymogen granule size and complexity, both parameters showing similar characteristics to those of the controls at day +7 after adrenalectomy. However, the percentage of zymogen granules within the R1 and R2 populations was clearly different from that of controls since the R2 population was much more numerous (11·25 ± 0·75% and 15·25 ± 1·15% (adrenalectomized rats at days +3 and +7 respectively) versus 4·45 ± 0·24% (controls)). An increase in the content of amylase per DNA was observed in adrenalectomized rats at day +1 although this transient effect cannot be related to glucocorticoid deprivation because it was also observed in sham-operated rats (day +1). However, a significant reduction, nearly 64%, in the amylase content/DNA ratio is produced by the absence of glucocorticoids 7 days after adrenalectomy and this is associated with a reduction in the content of amylase in each individual zymogen granule which reaches a minimum 3 days after adrenalectomy. It should be noted that, despite this, the enzyme concentration in each granule remains constant as there is a parallel decrease in the zymogen granule amylase content and size. Journal of Endocrinology (1995) 147, 431–440

Using dual-laser flow cytometry for monitoring phytoplankton composition and integrity

1995 ◽  
Author(s):  
Harald Schaefer ◽  
Wolfgang Beisker ◽  
Christian Steinberg
Keyword(s):  
Flow Cytometry ◽  
Phytoplankton Composition ◽  
Dual Laser

scholarly journals Detection of silver nanoparticles in cells by flow cytometry using light scatter and far-red fluorescence

2013 ◽  
pp. n/a-n/a ◽  
Author(s):  
R. M. Zucker ◽  
K. M. Daniel ◽  
E. J. Massaro ◽  
S. J. Karafas ◽  
L. L. Degn ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Silver Nanoparticles ◽  
Light Scatter ◽  
Red Fluorescence ◽  
In Cells

Diagnostic Utility of Flow Cytometry in Myelodysplastic Syndrome: A Retrospective Validation From a Single Centre.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4846-4846
Author(s):  
Pervin Topcuoglu ◽  
Klara Dalva ◽  
Sule Mine Bakanay ◽  
Sinem Civriz Bozdag ◽  
Onder Arslan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Myelodysplastic Syndrome ◽  
Antigen Expression ◽  
Pathological Examination ◽  
Light Scatter ◽  
Myeloid Lineage ◽  
Hematopoietic Stem ◽  
Altered Expression ◽  
Flow Cytometric

Abstract Abstract 4846 Myelodysplastic syndrome (MDS) is heterogeneous clonal hematopoietic stem cell disorder characterized by cytopenia(s) and dysplasia in one or more cell lineage. Though flow cytometry (FCM) is an important diagnostic tool in hematopoietic cell disorders, a prominent immunophenotyping feature in MDS may not be determined. In this study, we retrospectively evaluated flow cytometric features of bone marrow samples diagnosed as MDS with clinical and hematological findings. Patients-Method Between Feb 2004 and March 2009, flow cytometric parameters of 73 patients (M/F: 50/23) with MDS were re-analyzed. Median age was 59 years (17-89 ys). Our general principles are to evaluate quality of bone marrow samples, to determine proportion of the cells and features of their light scatter, and to give percentage of the blast. When detected a finding of dysplasia in the first analysis, the second step includes the determination of the maturation of the cells and the presence of the aberrant antigen expression. Results The samples were interpreted as MDS in % 76.7, MDS-RAEB-1 or RAEB-2 in %16.4, myeloproliferative disorder in %1.4 and non-diagnostic in %6.8 of the cases by flow cytometric examination. We detected variable degrees of hypogranulation in myeloid lineage in %82.2 of the samples by the light scatter features of the cells: 85% of severe and 15% of moderate or mild hypogranulation. The ratio of myeloid and lymphoid was changing from 0.3 to 17.5 (median 2). The decreasing of this ratio (<1) was observed in 19.4% of the samples. We detected altered expression of mature granulocyte. These included decreasing or lack of expression in CD15 45/73 (61.4%), CD13 38/70 (54.3%), CD16 53/67 (79.1%), CD11b 51/71 (71.8%), CD24 44/69 (65.2%), CD10 23/72 (31.9%) and MPO 14/72 (19.4%). Besides, bright expression of CD33 in 53.5% of the samples was observed. CD36 and CD56 in myeloid lineage were co-expressed in about 50 % of the samples. In 80.8 % of the samples dysplasia in erythroid compartment could be evaluated: Expression of CD71 according to glycophorin A (ratio <1) was decreased in 23.7 %. When we made similar analysis in the samples without RAEB-1 and -2 as pathological examination of bone marrow, 13.4 % of the samples could not be evaluated in favor of dysplasia. Of the samples with dysplasia hypogranulation, aberrant antigen expression of myeloid lineage and eryhtroid dysplasia were observed in 92.1%, 34.1% and 31.5%, respectively. In conclusion, FCM events may help to the differantial diagnosis of MDS especially when combining with clinical events. Improving of the analysis by focusing on the blast characteristics may be a standard approach to evaluate for low risk MDS. Disclosures No relevant conflicts of interest to declare.

Live-cell assay for detection of apoptosis by dual-laser flow cytometry using Hoechst 33342 and 7-amino-actinomycin D

2007 ◽  
Vol 2 (1) ◽  
pp. 187-190 ◽  
Author(s):  
Ingrid Schmid ◽  
Christel Uittenbogaart ◽  
Beth D Jamieson
Keyword(s):  
Flow Cytometry ◽  
Actinomycin D ◽  
Live Cell ◽  
Hoechst 33342 ◽  
Cell Assay ◽  
Dual Laser
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