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Generation of cell type-specific monoclonal antibodies for the planarian and optimization of sample processing for immunolabeling

Overview of attention for article published in BMC Developmental Biology, December 2014
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About this Attention Score

  • In the top 25% of all research outputs scored by Altmetric
  • Among the highest-scoring outputs from this source (#23 of 218)
  • High Attention Score compared to outputs of the same age (85th percentile)
  • Good Attention Score compared to outputs of the same age and source (68th percentile)

Mentioned by

blogs
1 blog
twitter
2 tweeters
facebook
1 Facebook page

Citations

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25 Dimensions

Readers on

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67 Mendeley
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Title
Generation of cell type-specific monoclonal antibodies for the planarian and optimization of sample processing for immunolabeling
Published in
BMC Developmental Biology, December 2014
DOI 10.1186/s12861-014-0045-6
Pubmed ID
Authors

David J Forsthoefel, Forrest A Waters, Phillip A Newmark

Abstract

BackgroundEfforts to elucidate the cellular and molecular mechanisms of regeneration have required the application of methods to detect specific cell types and tissues in a growing cohort of experimental animal models. For example, in the planarian Schmidtea mediterranea, substantial improvements to nucleic acid hybridization and electron microscopy protocols have facilitated the visualization of regenerative events at the cellular level. By contrast, immunological resources have been slower to emerge. Specifically, the repertoire of antibodies recognizing planarian antigens remains limited, and a more systematic approach is needed to evaluate the effects of processing steps required during sample preparation for immunolabeling.ResultsTo address these issues and to facilitate studies of planarian digestive system regeneration, we conducted a monoclonal antibody (mAb) screen using phagocytic intestinal cells purified from the digestive tracts of living planarians as immunogens. This approach yielded ten antibodies that recognized intestinal epitopes, as well as markers for the central nervous system, musculature, secretory cells, and epidermis. In order to improve signal intensity and reduce non-specific background for a subset of mAbs, we evaluated the effects of fixation and other steps during sample processing. We found that fixative choice, treatments to remove mucus and bleach pigment, as well as methods for tissue permeabilization and antigen retrieval profoundly influenced labeling by individual antibodies. These experiments led to the development of a step-by-step workflow for determining optimal specimen preparation for labeling whole planarians as well as unbleached histological sections.ConclusionsWe generated a collection of monoclonal antibodies recognizing the planarian intestine and other tissues; these antibodies will facilitate studies of planarian tissue morphogenesis. We also developed a protocol for optimizing specimen processing that will accelerate future efforts to generate planarian-specific antibodies, and to extend functional genetic studies of regeneration to post-transcriptional aspects of gene expression, such as protein localization or modification. Our efforts demonstrate the importance of systematically testing multiple approaches to species-specific idiosyncracies, such as mucus removal and pigment bleaching, and may serve as a template for the development of immunological resources in other emerging model organisms.

Twitter Demographics

The data shown below were collected from the profiles of 2 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 67 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Spain 1 1%
Germany 1 1%
Unknown 65 97%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 15 22%
Student > Master 8 12%
Student > Bachelor 7 10%
Professor > Associate Professor 7 10%
Researcher 6 9%
Other 12 18%
Unknown 12 18%
Readers by discipline Count As %
Agricultural and Biological Sciences 24 36%
Biochemistry, Genetics and Molecular Biology 21 31%
Engineering 2 3%
Medicine and Dentistry 2 3%
Immunology and Microbiology 2 3%
Other 6 9%
Unknown 10 15%

Attention Score in Context

This research output has an Altmetric Attention Score of 9. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 04 October 2015.
All research outputs
#980,108
of 8,004,786 outputs
Outputs from BMC Developmental Biology
#23
of 218 outputs
Outputs of similar age
#34,560
of 239,714 outputs
Outputs of similar age from BMC Developmental Biology
#5
of 16 outputs
Altmetric has tracked 8,004,786 research outputs across all sources so far. Compared to these this one has done well and is in the 87th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 218 research outputs from this source. They receive a mean Attention Score of 3.3. This one has done well, scoring higher than 88% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 239,714 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 85% of its contemporaries.
We're also able to compare this research output to 16 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 68% of its contemporaries.