Non-Linear Optical Microscopy Sheds Light on Cardiovascular Disease
Publication Date
February 07, 2013
Journal
PLOS ONE
Authors
Valentina Caorsi, Christopher Toepfer, Markus B. Sikkel, Alexander R. Lyon, et al
Volume
8
Issue
2
Pages
e56136
DOI
https://dx.plos.org/10.1371/journal.pone.0056136
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0056136
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/23409139
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3567079
Europe PMC
http://europepmc.org/abstract/MED/23409139
Web of Science
000315157200113
Scopus
84873593338
Mendeley
http://www.mendeley.com/research/nonlinear-optical-microscopy-sheds-light-cardiovascular-disease
Events
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Mendeley | Further Information

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Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/483829", "https://ndownloader.figshare.com/files/483832", "https://ndownloader.figshare.com/files/483833"], "description"=>"<div><p>Many cardiac diseases have been associated with increased fibrosis and changes in the organization of fibrillar collagen. The degree of fibrosis is routinely analyzed with invasive histological and immunohistochemical methods, giving a limited and qualitative understanding of the tissue's morphological adaptation to disease. Our aim is to quantitatively evaluate the increase in fibrosis by three-dimensional imaging of the collagen network in the myocardium using the non-linear optical microscopy techniques Two-Photon Excitation microscopy (TPE) and Second Harmonic signal Generation (SHG). No sample staining is needed because numerous endogenous fluorophores are excited by a two-photon mechanism and highly non-centrosymmetric structures such as collagen generate strong second harmonic signals. We propose for the first time a 3D quantitative analysis to carefully evaluate the increased fibrosis in tissue from a rat model of heart failure post myocardial infarction. We show how to measure changes in fibrosis from the backward SHG (B<sub>SHG</sub>) alone, as only backward-propagating SHG is accessible for true <em>in vivo</em> applications. A 5-fold increase in collagen I fibrosis is detected in the remote surviving myocardium measured 20 weeks after infarction. The spatial distribution is also shown to change markedly, providing insight into the morphology of disease progression.</p> </div>", "links"=>[], "tags"=>["non-linear", "optical", "microscopy", "sheds", "cardiovascular", "disease"], "article_id"=>156212, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.s001", "https://dx.doi.org/10.1371/journal.pone.0056136.s002", "https://dx.doi.org/10.1371/journal.pone.0056136.s003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Non_Linear_Optical_Microscopy_Sheds_Light_on_Cardiovascular_Disease__/156212", "title"=>"Non-Linear Optical Microscopy Sheds Light on Cardiovascular Disease", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-02-07 01:43:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/492669"], "description"=>"<p>Standard histology sections are stained with picrosirius red to highlight collagen in tissues from myocardial infarction rat model (a and b), from age-match control in c, showing the better signal to noise ratio for SHG light in discriminating collagen. In d, the spectral characterization is presented to demonstrate that the collected signals are SHG (magenta) by observing the shift in peak intensity with that of the excitation wavelength and autofluorescence (green). Scale bar 50 µm in a, 100 µm in b and c.</p>", "links"=>[], "tags"=>["autofluorescence"], "article_id"=>163187, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_B_SHG_and_autofluorescence_characterisation_/163187", "title"=>"B<sub>SHG</sub> and autofluorescence characterisation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:53:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/492791"], "description"=>"<p>Skeletal fibres (a, c, e, g, circle symbols in the graph) and cardiac trabeculae (b, d, f, h, squared symbols in the graph) imaged at increasing excitation power ranging from 10 mW up to 70 mW at 900 nm. In i), filled squared symbols are related to collagen-B<sub>SHG</sub>, while open squared symbols are related to myosin-B<sub>SHG</sub>, obtained from analyzing region of interest showing only collagen fibrosis or sarcomeric proteins respectively, as these are well spatially separated. By utilizing low power (10–20 mW) myosin-B<sub>SHG</sub> can be neglected, thus ensuring the collection of only collagen-B<sub>SHG</sub>. Scale bar 100 µm.</p>", "links"=>[], "tags"=>["physiology", "biophysics"], "article_id"=>163312, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Myosin_B_SHG_detection_/163312", "title"=>"Myosin-B<sub>SHG</sub> detection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:55:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/492907"], "description"=>"<p>B<sub>SHG</sub> from rat tail samples has been collected at different NaCl concentrations compared to relaxing solution. From 50 mM NaCl the B<sub>SHG</sub> reaches a plateau. Measurements performed in relaxing solution fall in the same range of intensity ensuring that in the present experimental conditions we maximize the backward light collection. Images above symbols are representative for that concentration.</p>", "links"=>[], "tags"=>["dependence", "ionic"], "article_id"=>163426, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_B_SHG_dependence_on_ionic_strength_/163426", "title"=>"B<sub>SHG</sub> dependence on ionic strength.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:57:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/493019"], "description"=>"<p>Representative 3D optical sections (xy and orthogonal view xz, yz) of a trabecula in a) and a rat tail in b); green autofluorescence, fire color scale B<sub>SHG</sub>. Scale bar, 25 µm. In c) B<sub>SHG</sub> intensity versus sample depth is presented for rat tail samples (black), cardiac trabeculae (dark grey) and permeabilised skeletal fibres (light grey). Intensity profiles have been normalized to the rat tail samples as they represent the upper bound of B<sub>SHG</sub> detection. On the contrary permeabilised skeletal fibres show virtually no B<sub>SHG</sub> (2%) thus representing the lower bound of detection. Healthy cardiac samples show a low but measurable B<sub>SHG</sub>, thus ensuring a large range of detection for possible increases in collagen content in diseased samples.</p>", "links"=>[], "tags"=>["detection"], "article_id"=>163541, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Upper_and_lower_B_SHG_detection_range_/163541", "title"=>"Upper and lower B<sub>SHG</sub> detection range.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:59:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/493115"], "description"=>"<p>3D (shown as xy view and orthogonal views xz, yz) autofluorescence a) and B<sub>SHG</sub> b) from a MI sample. The region of interest selected by thresholding the autofluorescence to define the sample edges is highlighted in green in a); the very same region is applied to the B<sub>SHG</sub> images, highlighted in yellow in b); c) intensity profile along the thickness of the sample, MI (dark red, red, orange), AMC black and grey; d) Average intensity increase (MI/AMC) obtained by integrating B<sub>SHG</sub> intensity over 10 µm z-steps.</p>", "links"=>[], "tags"=>["physiology", "biophysics"], "article_id"=>163637, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Intensity_Analysis_/163637", "title"=>"Intensity Analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:00:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/493219"], "description"=>"<p>3D (shown as xy view and orthogonal views xz, yz) autofluorescence a) and B<sub>SHG</sub> b) from a MI sample. The region of interest selected by thresholding the autofluorescence to define the total volume of the sample is highlighted in green in a); the region of interest selected by thresholding the B<sub>SHG</sub> to define the volume occupied by collagen is highlighted in yellow in b); c) the discretized area vs depth is plotted, black and red is from autofluorescence (AMC and MI samples respectively) grey and orange from B<sub>SHG</sub> (AMC and MI sample respectively); the curves have been normalized so that the integral over black and red is 1, therefore representing the total volume, and over grey and orange is the fraction of volume occupied by collagen; d) average collagen presence evaluated for MI (n = 8) compared to AMC (n = 8) animals.</p>", "links"=>[], "tags"=>["physiology", "biophysics"], "article_id"=>163734, "categories"=>["Physiology", "Biophysics"], "users"=>["Valentina Caorsi", "Christopher Toepfer", "Markus B. Sikkel", "Alexander R. Lyon", "Ken MacLeod", "Mike A. Ferenczi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0056136.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Volume_Analysis_/163734", "title"=>"Volume Analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:02:14"}

PMC Usage Stats | Further Information

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Relative Metric

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