Automated image analysis to quantify the subnuclear organization of transcriptional coregulatory protein complexes in living cell populations

2004 ◽  
Author(s):  
Ty C. Voss ◽  
Ignacio A. Demarco ◽  
Cynthia F. Booker ◽  
Richard N. Day
Keyword(s):  
Image Analysis ◽  
Living Cell ◽  
Protein Complexes ◽  
Automated Image Analysis ◽  
Cell Populations ◽  
Coregulatory Protein

scholarly journals Bright-Field Microscopy Visualization of Proteins and Protein Complexes by In Situ Proximity Ligation with Peroxidase Detection

2010 ◽  
Vol 56 (1) ◽  
pp. 99-110 ◽  
Author(s):  
Agata Zieba ◽  
Carolina Wählby ◽  
Fredrik Hjelm ◽  
Lee Jordan ◽  
Jonathan Berg ◽  
...  
Keyword(s):  
Image Analysis ◽  
Protein Complexes ◽  
Automated Image Analysis ◽  
Proximity Ligation Assay ◽  
Bright Field ◽  
Proximity Ligation ◽  
Tissue Sections ◽  
Dna Molecule ◽  
Bright Field Microscopy

Abstract Background: The in situ proximity ligation assay (PLA) allows a protein or protein complex to be represented as an amplifiable DNA molecule. Recognition is mediated by proximity probes consisting of antibodies coupled with oligonucleotides. Upon dual binding of the proximity probes, the oligonucleotides direct the formation of a circular DNA molecule, which is then amplified by rolling-circle replication. The localized concatemeric product is then detected with fluorescent probes. The in situ PLA enables localized detection of individual native proteins or interacting protein pairs in fixed cells or tissue sections, thus providing an important tool for basic and clinical research. Methods: We used horseradish peroxidase (HRP)-conjugated oligonucleotides to couple in situ PLA with enzymatic visualization of the localized detection event. Results: We demonstrate the detection of protein complexes, both in cells and in tissue sections, and show that we can quantify the complexes with image-analysis software specially developed for recognizing HRP signals in bright-field microscopy images. We show that fluorescence and HRP signals produce equivalent results, both in cultured cells and in tissue samples. Conclusions: The combination of in situ PLA with bright-field detection and automated image analysis allows the signals present to be counted in an automated fashion and thus provides a sensitive and specific method for quantification of proteins and protein complexes with bright-field microscopy. With this approach, in situ PLA can be used without the requirement for expensive fluorescence microscopes, thereby avoiding problems with nonspecific fluorescence while maintaining compatibility with conventional histologic staining.

Automated Image Analysis Of Nuclear And Cytoplasmic Changes In Exfoliated Cells From Tracheas Treated With Carcinogen

1977 ◽  
Vol 35 ◽  
pp. 220-221
Author(s):  
S.F. Stinson ◽  
J.C. Lilga ◽  
M.B. Sporn
Keyword(s):  
Image Analysis ◽  
Pattern Analysis ◽  
Relative Proportion ◽  
Automated Image Analysis ◽  
Image Analyzer ◽  
Cross Sectional ◽  
Exfoliated Cells ◽  
Neoplastic Lesions ◽  
Analysis System ◽  
Morphologic Criterion

Increased nuclear size, resulting in an increase in the relative proportion of nuclear to cytoplasmic sizes, is an important morphologic criterion for the evaluation of neoplastic and pre-neoplastic cells. This paper describes investigations into the suitability of automated image analysis for quantitating changes in nuclear and cytoplasmic cross-sectional areas in exfoliated cells from tracheas treated with carcinogen.Neoplastic and pre-neoplastic lesions were induced in the tracheas of Syrian hamsters with the carcinogen N-methyl-N-nitrosourea. Cytology samples were collected intra-tracheally with a specially designed catheter (1) and stained by a modified Papanicolaou technique. Three cytology specimens were selected from animals with normal tracheas, 3 from animals with dysplastic changes, and 3 from animals with epidermoid carcinoma. One hundred randomly selected cells on each slide were analyzed with a Bausch and Lomb Pattern Analysis System automated image analyzer.

Progress In The Practical Application Of Automated Image Analysis To Tem Negatives

1977 ◽  
Vol 35 ◽  
pp. 214-217
Author(s):  
F. A. Heckman ◽  
E. Redman ◽  
J.E. Connolly
Keyword(s):  
Image Analysis ◽  
Image Quality ◽  
Exposure Time ◽  
Image Contrast ◽  
Automated Image Analysis ◽  
Practical Application ◽  
Factors Affecting ◽  
High Image Quality ◽  
Qualitative Evidence ◽  
Comprehensive Study

In our initial publication on this subject1) we reported results demonstrating that contrast is the most important factor in producing the high image quality required for reliable image analysis. We also listed the factors which enhance contrast in order of the experimentally determined magnitude of their effect. The two most powerful factors affecting image contrast attainable with sheet film are beam intensity and KV. At that time we had only qualitative evidence for the ranking of enhancing factors. Later we carried out the densitometric measurements which led to the results outlined below.Meaningful evaluations of the cause-effect relationships among the considerable number of variables in preparing EM negatives depend on doing things in a systematic way, varying only one parameter at a time. Unless otherwise noted, we adhered to the following procedure evolved during our comprehensive study:Philips EM-300; 30μ objective aperature; magnification 7000- 12000X, exposure time 1 second, anti-contamination device operating.

Interfacing of a TEM/STEM System to a Computer

1981 ◽  
Vol 39 ◽  
pp. 250-251
Author(s):  
P. Hagemann
Keyword(s):  
Image Analysis ◽  
Transmission Rate ◽  
Analytical Electron Microscopy ◽  
Signal Beam ◽  
Automated Image Analysis ◽  
Beam Deflection ◽  
External Control ◽  
Computer Input ◽  
Instrument Control ◽  
Data Acquisition And Processing

The use of computers in the analytical electron microscopy today shows three different trends (1) automated image analysis with dedicated computer systems, (2) instrument control by microprocessors and (3) data acquisition and processing e.g. X-ray or EEL Spectroscopy.While image analysis in the T.E.M. usually needs a television chain to get a sequential transmission suitable as computer input, the STEM system already has this necessary facility. For the EM400T-STEM system therefore an interface was developed, that allows external control of the beam deflection in TEM as well as the control of the STEM probe and video signal/beam brightness on the STEM screen.The interface sends and receives analogue signals so that the transmission rate is determined by the convertors in the actual computer periphery.

Electron Microscopy and image analysis of nucleo-protein complexes

1994 ◽  
Vol 52 ◽  
pp. 88-89
Author(s):  
E. H. Egelman ◽  
X. Yu
Keyword(s):  
Electron Microscopy ◽  
Image Analysis ◽  
Normal Cell ◽  
Genetic Recombination ◽  
Protein Complexes ◽  
Chromosomal Rearrangements ◽  
Reca Protein ◽  
E Coli ◽  
Helical Polymer ◽  
Specific Protease

The RecA protein of E. coli has been shown to mediate genetic recombination, regulate its own synthesis, control the expression of other genes, act as a specific protease, form a helical polymer and have an ATPase activity, among other observed properties. The unusual filament formed by the RecA protein on DNA has not previously been shown to exist outside of bacteria. Within this filament, the 36 Å pitch of B-form DNA is extended to about 95 Å, the pitch of the RecA helix. We have now establishedthat similar nucleo-protein complexes are formed by bacteriophage and yeast proteins, and availableevidence suggests that this structure is universal across all of biology, including humans. Thus, understanding the function of the RecA protein will reveal basic mechanisms, in existence inall organisms, that are at the foundation of general genetic recombination and repair.Recombination at this moment is assuming an importance far greater than just pure biology. The association between chromosomal rearrangements and neoplasms has become stronger and stronger, and these rearrangements are most likely products of the recombinatory apparatus of the normal cell. Further, damage to DNA appears to be a major cause of cancer.

scholarly journals Author Correction: Efficient microbial colony growth dynamics quantification with ColTapp, an automated image analysis application

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Julian Bär ◽  
Mathilde Boumasmoud ◽  
Roger D. Kouyos ◽  
Annelies S. Zinkernagel ◽  
Clément Vulin
Keyword(s):  
Image Analysis ◽  
Growth Dynamics ◽  
Colony Growth ◽  
Automated Image Analysis ◽  
Analysis Application

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Scoring of radiation-induced microuclei in cytokinesis-blocked human lymphocytes by automated image analysis

Cytometry ◽  
1994 ◽  
Vol 17 (2) ◽  
pp. 119-127 ◽  
Author(s):  
F. Verhaegen ◽  
A. Vral ◽  
J. Seuntjens ◽  
N. W. Schipper ◽  
L. de Ridder ◽  
...  
Keyword(s):  
Image Analysis ◽  
Human Lymphocytes ◽  
Automated Image Analysis ◽  
Radiation Induced

An automated image analysis platform for the study of weakly -adhered cells

Biofouling ◽  
2021 ◽  
pp. 1-10
Author(s):  
Zhijing Wan ◽  
Ben T. MacVicar ◽  
Shea Wyatt ◽  
Diana E. Varela ◽  
Rajkumar Padmawar ◽  
...  
Keyword(s):  
Image Analysis ◽  
Automated Image Analysis ◽  
Analysis Platform

scholarly journals Yeast Morphology Assessment through Automated Image Analysis during Fermentation

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 44
Author(s):  
Mario Guadalupe-Daqui ◽  
Mandi Chen ◽  
Katherine A. Thompson-Witrick ◽  
Andrew J. MacIntosh
Keyword(s):  
Image Analysis ◽  
High Stress ◽  
Cross Sectional Area ◽  
Automated Image Analysis ◽  
Yeast Cells ◽  
Morphological Properties ◽  
Cell Area ◽  
Sectional Area ◽  
Cross Sectional ◽  
Industrial Fermentation

The kinetics and success of an industrial fermentation are dependent upon the health of the microorganism(s) responsible. Saccharomyces sp. are the most commonly used organisms in food and beverage production; consequently, many metrics of yeast health and stress have been previously correlated with morphological changes to fermentations kinetics. Many researchers and industries use machine vision to count yeast and assess health through dyes and image analysis. This study assessed known physical differences through automated image analysis taken throughout ongoing high stress fermentations at various temperatures (30 °C and 35 °C). Measured parameters included sugar consumption rate, number of yeast cells in suspension, yeast cross-sectional area, and vacuole cross-sectional area. The cell morphological properties were analyzed automatically using ImageJ software and validated using manual assessment. It was found that there were significant changes in cell area and ratio of vacuole to cell area over the fermentation. These changes were temperature dependent. The changes in morphology have implications for rates of cellular reactions and efficiency within industrial fermentation processes. The use of automated image analysis to quantify these parameters is possible using currently available systems and will provide additional tools to enhance our understanding of the fermentation process.

scholarly journals Alignment of Au nanorods along de novo designed protein nanofibers studied with automated image analysis

Soft Matter ◽  
2021 ◽  
Author(s):  
Muammer Y. Yaman ◽  
Kathryn N. Guye ◽  
Maxim Ziatdinov ◽  
Hao Shen ◽  
David Baker ◽  
...  
Keyword(s):  
Image Analysis ◽  
De Novo ◽  
Automated Image Analysis ◽  
Analysis Tool ◽  
Au Nanorods

In this study, we focus on exploring the directional assembly of anisotropic Au nanorods along de novo designed 1D protein nanofiber templates using automated image analysis tool.

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