Flow Cytometric Measurement of Specific DNA and RNA Sequences

1989 ◽  
pp. 275-301 ◽  
Author(s):  
J.G.J. BAUMAN ◽  
D. PINKEL ◽  
B.J. TRASK ◽  
M. VAN DER PLOEG
Keyword(s):  
Rna Sequences ◽  
Dna And Rna ◽  
Flow Cytometric ◽  
Flow Cytometric Measurement

High-resolution nucleic acid sequence mapping via in situ hybridization at the Electron Microscope level

1995 ◽  
Vol 53 ◽  
pp. 752-753
Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch
Keyword(s):  
Nucleic Acid ◽  
Interphase Nucleus ◽  
Genome Mapping ◽  
Precise Location ◽  
Electron Microscope Level ◽  
Rna Sequences ◽  
Fundamental Interest ◽  
Whole Cells ◽  
Dna And Rna

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.

scholarly journals Interleukin 8 Elicits Rapid Physiological Changes in Neutrophils That Are Altered by Inflammatory Conditions

2021 ◽  
pp. 1-17
Author(s):  
Stefan Bernhard ◽  
Stefan Hug ◽  
Alexander Elias Paul Stratmann ◽  
Maike Erber ◽  
Laura Vidoni ◽  
...  
Keyword(s):  
Systemic Inflammation ◽  
Severe Trauma ◽  
Induced Response ◽  
Neutrophil Granulocytes ◽  
Inflammatory Conditions ◽  
Flow Cytometric ◽  
Time Flow ◽  
Flow Cytometric Measurement ◽  
Detailed Kinetics

A sufficient response of neutrophil granulocytes stimulated by interleukin (IL)-8 is vital during systemic inflammation, for example, in sepsis or severe trauma. Moreover, IL-8 is clinically used as biomarker of inflammatory processes. However, the effects of IL-8 on cellular key regulators of neutrophil properties such as the intracellular pH (pH<sub>i</sub>) in dependence of ion transport proteins and during inflammation remain to be elucidated. Therefore, we investigated in detail the fundamental changes in pH<sub>i</sub>, cellular shape, and chemotactic activity elicited by IL-8. Using flow cytometric methods, we determined that the IL-8-induced cellular activity was largely dependent on specific ion channels and transporters, such as the sodium-proton exchanger 1 (NHE1) and non-NHE1-dependent sodium flux. Exposing neutrophils in vitro to a proinflammatory micromilieu with N-formyl-Met-Leu-Phe, LPS, or IL-8 resulted in a diminished response regarding the increase in cellular size and pH. The detailed kinetics of the reduced reactivity of the neutrophil granulocytes could be illustrated in a near-real-time flow cytometric measurement. Last, the LPS-mediated impairment of the IL-8-induced response in neutrophils was confirmed in a translational, animal-free human whole blood model. Overall, we provide novel mechanistic insights for the interaction of IL-8 with neutrophil granulocytes and report in detail about its alteration during systemic inflammation.

Tracking DNA and RNA Sequences at High Resolution

2013 ◽  
pp. 343-366 ◽  
Author(s):  
Dušan Cmarko ◽  
Anna Ligasová ◽  
Karel Koberna
Keyword(s):  
High Resolution ◽  
Rna Sequences ◽  
Dna And Rna

scholarly journals STANDARDIZATION OF FLOW CYTOMETRIC MEASUREMENT OF HLA-DR MONOCYTES 53

1997 ◽  
Vol 41 (5) ◽  
pp. 752-752
Author(s):  
F Debonera ◽  
A Germenis ◽  
G Paterakis ◽  
S Fotopoulos ◽  
M Xanthou ◽  
...  
Keyword(s):  
Flow Cytometric ◽  
Hla Dr ◽  
Flow Cytometric Measurement

Hyperdiploidy and chromosomal rearrangements define the anaplastic variant of Wilms' tumor.

1986 ◽  
Vol 4 (6) ◽  
pp. 975-981 ◽  
Author(s):  
E C Douglass ◽  
A T Look ◽  
B Webber ◽  
D Parham ◽  
J A Wilimas ◽  
...  
Keyword(s):  
Dna Content ◽  
Wilms Tumor ◽  
Chromosomal Rearrangements ◽  
Chromosome Analysis ◽  
Histologic Subtype ◽  
Flow Cytometric ◽  
Kaplan Meier ◽  
Flow Cytometric Measurement ◽  
Flow Cytometric Study ◽  
Diploid Cells

Flow cytometric measurement of the DNA content of Wilms' tumor cells revealed a striking correspondence with the histologic subtype and treatment outcome. In the 48 cases studied, a hyperdiploid DNA content ranging from 1.7 to 3.2 times the result for normal diploid cells distinguished all but one of the ten anaplastic tumors. Lower values, from 1.0 to 1.4 times the diploid DNA content, characterized the nonanaplastic specimens. By Kaplan-Meier analysis, the probability of achieving 3 years of relapse-free survival was significantly lower in the group with higher DNA content (0.42 v 0.87, P less than .01). Analysis of banded chromosomes for a subset of 22 patients contributed important information beyond the flow cytometric study. Cases of anaplasia associated with poorer responses to therapy showed numerous complex translocations, whereas all others lacked such changes. By combining flow cytometric techniques and conventional methods of chromosome analysis, it should be possible to identify those patients with Wilms' tumor who are most likely to fail therapy. The biologic implication of these findings is that the development of clinical drug resistance in Wilms' tumor is a result of the genetic instability of the malignant clone.

scholarly journals Electrolytic degradation of DNA fluorochromes during flow cytometric measurement of electronic cell volume.

1980 ◽  
Vol 28 (4) ◽  
pp. 330-334 ◽  
Author(s):  
O Alabaster ◽  
D L Glaubiger ◽  
V T Hamilton ◽  
S A Bentley ◽  
S E Shackney ◽  
...  
Keyword(s):  
Cell Volume ◽  
Dna Content ◽  
Chemical Degradation ◽  
Quantum Yields ◽  
Light Scatter ◽  
Electrolytic Production ◽  
Flow Cytometric ◽  
Chlorine Gas ◽  
Electrode Polarity ◽  
Flow Cytometric Measurement

Changes in flow cytometric measurement of DNA content can result from electrolytic chemical degradation of mithramycin, ethidium bromide, and propidium iodide during simultaneous measurement of electronic cell volume. Bench electrolysis also degrades these fluorochromes without changing the quantum yields, even when they are complexed to DNA. In the flow cytometer, electrolytic production of chlorine at the anode is the probable cause of this degradation, since exposure of these fluorochromes to chlorine gas produces the same effect. It is therefore advisable to measure the DNA content distribution alone before simultaneously measuring the DNA content and the electronic cell volume. If unavoidable effects on the DNA distribution are present, narrow forward-angle light scatter should be used as the cell size indicator during dual parameter measurements. Modifying instrument design by reversing electrode polarity might eliminate this problem.

P1-351: FLOW CYTOMETRIC MEASUREMENT OF MICROPARTICLES IN HUMAN CEREBROSPINAL FLUID OF NEURODEGENERATIVE DISORDERS

2014 ◽  
Vol 10 ◽  
pp. P442-P442
Author(s):  
Yue Yang ◽  
Elaine R. Peskind ◽  
Eiron Cudaback ◽  
Angela M. Wilson ◽  
Thomas J. Montine ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Neurodegenerative Disorders ◽  
Flow Cytometric ◽  
Human Cerebrospinal Fluid ◽  
Flow Cytometric Measurement
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