scholarly journals Application of Digital Image Analysis and Flow Cytometry To Enumerate Marine Viruses Stained with SYBR Gold

2001 ◽  
Vol 67 (2) ◽  
pp. 539-545 ◽  
Author(s):  
Feng Chen ◽  
Jing-rang Lu ◽  
Brian J. Binder ◽  
Ying-chun Liu ◽  
Robert E. Hodson
Keyword(s):  
Flow Cytometry ◽  
Image Analysis ◽  
Digital Image ◽  
Digital Image Analysis ◽  
Flow Cytometric Analysis ◽  
Epifluorescence Microscopy ◽  
Flow Cytometric ◽  
Viral Particles ◽  
Sybr Gold ◽  
Nucleic Acid Stain

ABSTRACT A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of SYBR Gold-stained viruses was about twice that of SYBR Green I-stained viruses. Digital images of SYBR Gold-stained viral particles were processed to enumerate the concentration of viral particles by using digital image analysis software. Estimates of viral concentration based on digitized images were 1.3 times higher than those based on direct counting by epifluorescence microscopy. Direct epifluorescence counts of SYBR Gold-stained viral particles were in turn about 1.34 times higher than those estimated by the transmission electron microscope method. Bacteriophage lysates stained with SYBR Gold formed a distinct population in flow cytometric signatures. Flow cytometric analysis revealed at least four viral subpopulations for a Lake Erie sample and two subpopulations for a Georgia coastal sample. Flow cytometry-based viral counts for various types of samples averaged 1.1 times higher than direct epifluorescence microscopic counts. The potential application of digital image analysis and flow cytometry for rapid and accurate measurement of viral abundance in aquatic environments is discussed.

Model-based frequency response characterization of a digital-image analysis system for epifluorescence microscopy

1992 ◽  
Vol 31 (8) ◽  
pp. 1083
Author(s):  
Rajeeb Hazra ◽  
Charles L. Viles ◽  
Stephen K. Park ◽  
Stephen E. Reichenbach ◽  
Michael E. Sieracki
Keyword(s):  
Image Analysis ◽  
Frequency Response ◽  
Digital Image ◽  
Digital Image Analysis ◽  
Epifluorescence Microscopy ◽  
Image Analysis System ◽  
Model Based ◽  
Analysis System ◽  
Digital Image Analysis System

“Decoding the Dots”: The ImageStream system (ISS) as a novel and powerful tool for flow cytometric analysis

2008 ◽  
Vol 3 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Ewa Zuba-Surma ◽  
Magdalena Kucia ◽  
Mariusz Ratajczak
Keyword(s):  
Flow Cytometry ◽  
Laser Scanning ◽  
Digital Image Analysis ◽  
Fluorescent Microscopy ◽  
Flow Cytometric Analysis ◽  
Cell Analysis ◽  
Photometric Analysis ◽  
Flow Cytometric ◽  
Novel Method ◽  
Laser Scanning Cytometer

AbstractThe aim of this article is to provide a brief review of the ImageStream system (ISS). The ISS technology was developed as a novel method for multiparameter cell analysis and subsequently as a supportive tool for flow cytometry (FC). ISS integrates the features of FC and fluorescent microscopy collecting images of acquired cells for offline digital image analysis. The article presents an overview of the main characteristics of ISS and a comparison between ISS, FC and the laser scanning cytometer (LSC). We reviewed ISS applications focusing on those involved in cellular phenotyping and provide our own experience with using ISS as a supportive tool to classical FC and demonstrate the compatibility between FC and ISS photometric analysis as well as the advantages of using ISS to confirm FC results.

scholarly journals Automation of human sperm cell analysis by flow cytometry

1997 ◽  
Vol 43 (5) ◽  
pp. 801-807 ◽  
Author(s):  
Fulvio Ferrara ◽  
Rita Daverio ◽  
Giuliano Mazzini ◽  
Pierangelo Bonini ◽  
Giuseppe Banfi
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Human Sperm ◽  
Sperm Analysis ◽  
Cell Counts ◽  
Flow Cytometric ◽  
Sperm Counts ◽  
The Mean ◽  
Nucleic Acid Stain ◽  
Automated Methods

Abstract Semen sample analysis is routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. Our aim was to automate sperm analysis with the use of flow cytometry for evaluation of cell counts and typing and with the use of a new membrane-permeant nucleic acid stain for evaluation of sperm viability. Statistical analysis of the comparison between manual and automated methods for sperm counts was performed by the Bland and Altman method; the mean difference was 0.243 × 106 sperms/mL. The precision of the flow cytometric analysis was evaluated with whole sperm; the between-run CV was 7.5% and the within-run CV was 2.5%. Data observed suggest that flow cytometric sperm analysis, with high precision and accuracy and low costs, can be proposed for routine use in clinical laboratories.

scholarly journals Chromosome Aberration Detection with Hybridized DNA Probes: Digital Image Analysis and Slit Scan Flow Cytometry

1989 ◽  
pp. 123-132 ◽  
Author(s):  
Christoph Cremer ◽  
Michael Hausmann ◽  
Eduardo Diaz ◽  
Jutta Hetzel ◽  
Jacob A. Aten ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Image Analysis ◽  
Chromosome Aberration ◽  
Digital Image ◽  
Digital Image Analysis ◽  
Dna Probes ◽  
Aberration Detection

scholarly journals Assessment and Interpretation of Bacterial Viability by Using the LIVE/DEAD BacLight Kit in Combination with Flow Cytometry

2007 ◽  
Vol 73 (10) ◽  
pp. 3283-3290 ◽  
Author(s):  
Michael Berney ◽  
Frederik Hammes ◽  
Franziska Bosshard ◽  
Hans-Ulrich Weilenmann ◽  
Thomas Egli
Keyword(s):  
Flow Cytometry ◽  
Outer Membrane ◽  
Shigella Flexneri ◽  
Epifluorescence Microscopy ◽  
E Coli ◽  
Flow Cytometric ◽  
Freshwater Bacteria ◽  
Serovar Typhimurium ◽  
Nucleic Acid Stain ◽  
Green Fluorescent

ABSTRACT The commercially available LIVE/DEAD BacLight kit is enjoying increased popularity among researchers in various fields of microbiology. Its use in combination with flow cytometry brought up new questions about how to interpret LIVE/DEAD staining results. Intermediate states, normally difficult to detect with epifluorescence microscopy, are a common phenomenon when the assay is used in flow cytometry and still lack rationale. It is shown here that the application of propidium iodide in combination with a green fluorescent total nucleic acid stain on UVA-irradiated cells of Escherichia coli, Salmonella enterica serovar Typhimurium, Shigella flexneri, and a community of freshwater bacteria resulted in a clear and distinctive flow cytometric staining pattern. In the gram-negative bacterium E. coli as well as in the two enteric pathogens, the pattern can be related to the presence of intermediate cellular states characterized by the degree of damage afflicted specifically on the bacterial outer membrane. This hypothesis is supported by the fact that EDTA-treated nonirradiated cells exhibit the same staining properties. On the contrary, this pattern was not observed in gram-positive Enterococcus faecalis, which lacks an outer membrane. Our observations add a new aspect to the LIVE/DEAD stain, which so far was believed to be dependent only on cytoplasmic membrane permeability.

Digital Image Analysis of the Larynx

2000 ◽  
Vol 10 (2) ◽  
pp. 7-9
Author(s):  
Yaser Natour ◽  
Christine Sapienza ◽  
Mark Schmalz ◽  
Savita Collins
Keyword(s):  
Image Analysis ◽  
Digital Image ◽  
Digital Image Analysis
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