Filter-Dense Multicolor Microscopy

Kijani, Siavash; Heyden, Maria; Levin, Malin; Borén, Jan; Fogelstrand, Per

doi:10.1371/journal.pone.0119499

Cited by 12 publications

(12 citation statements)

References 25 publications

(39 reference statements)

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“…To investigate a potential effect of the gut microbiota on IEC differentiation, we determined the cellular composition in ileum and proximal colon of GF and CR mice using immunostaining with a recently developed multi-channel setup [ 16 ] and an automated slide scanning and picture analysis pipeline. Overall, we found only minor alterations of IEC type abundance between GF and CR mice with slightly more Paneth and goblet cells in CR ileum (Figure 1 A).…”

Section: Resultsmentioning

confidence: 99%

“…HOECHST (Life Technologies, #H1399) was used as DNA counterstaining. Slides were scanned using Metafer automated slide scanner (MetaSystems, Altlussheim, Baden-Württemberg, Germany) and composite pictures analyzed using the Visiopharm Integrator System program [ 16 ]. Total area for each staining was determined for complete intestinal sections and expressed as percentage of epithelial area (E-cadherin).…”

Section: Methodsmentioning

confidence: 99%

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Site-specific programming of the host epithelial transcriptome by the gut microbiota

et al. 2015

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BackgroundThe intestinal epithelium separates us from the microbiota but also interacts with it and thus affects host immune status and physiology. Previous studies investigated microbiota-induced responses in the gut using intact tissues or unfractionated epithelial cells, thereby limiting conclusions about regional differences in the epithelium. Here, we sought to investigate microbiota-induced transcriptional responses in specific fractions of intestinal epithelial cells. To this end, we used microarray analysis of laser capture microdissection (LCM)-harvested ileal and colonic tip and crypt epithelial fractions from germ-free and conventionally raised mice and from mice during the time course of colonization.ResultsWe found that about 10% of the host’s transcriptome was microbially regulated, mainly including genes annotated with functions in immunity, cell proliferation, and metabolism. The microbial impact on host gene expression was highly site specific, as epithelial responses to the microbiota differed between cell fractions. Specific transcriptional regulators were enriched in each fraction. In general, the gut microbiota induced a more rapid response in the colon than in the ileum.ConclusionsOur study indicates that the microbiota engage different regulatory networks to alter host gene expression in a particular niche. Understanding host-microbiota interactions on a cellular level may facilitate signaling pathways that contribute to health and disease and thus provide new therapeutic strategies.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0614-4) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Site-specific programming of the host epithelial transcriptome by the gut microbiota

et al. 2015

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“…The PP was visualized using an Axioplan Carl Zeiss microscope set up for filter-dense multicolor microscopy. 194 Individual images for each filter were imported into ImageJ for preparation of composite images.…”

Section: Introductionmentioning

confidence: 99%

The regulation of gut mucosal IgA B-cell responses: recent developments

2017

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The majority of activated B cells differentiate into IgA plasma cells, with the gut being the largest producer of immunoglobulin in the body. Secretory IgA antibodies have numerous critical functions of which protection against infections and the role for establishing a healthy microbiota appear most important. Expanding our knowledge of the regulation of IgA B-cell responses and how effective mucosal vaccines can be designed are of critical importance. Here we discuss recent developments in the field that shed light on the uniqueness and complexity of mucosal IgA responses and the control of protective IgA responses in the gut, specifically.

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“…High‐resolution images were acquired using Metasystem automated slide scanner (MetaSystems, Altlussheim, Germany) equipped with SpectraSplit filter system for extended multicolor imaging (Kromnigon AB, Gothenburg, Sweden) . We used the Zeiss AxioImager.Z2 microscope; objective: Plan‐Apochromat 40×/1.4 oil objective (Carl Zeiss Microscopy, Jenna, Germany).…”

Section: Methodsmentioning

confidence: 99%

Endothelial repair is dependent on CD11c+ leukocytes to establish regrowing endothelial sheets with high cellular density

Holm

Levin

Alsén

et al. 2018

Journal of Leukocyte Biology

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Endothelial injury makes the vessel wall vulnerable to cardiovascular diseases. Injured endothelium regenerates by collective sheet migration, that is, the endothelial cells coordinate their motion and regrow as a sheet of cells with retained cell-cell contacts into the wounded area.Leukocytes appear to be involved in endothelial repair in vivo; however, little is known about their identity and role in the reparative sheet migration process. To address these questions, we developed a high-quality en face technique that enables visualizing of leukocytes and endothelial cells simultaneously following an endoluminal scratch wound injury of the mouse carotid artery. We discovered that regrowing endothelium forms a broad proliferative front accompanied by CD11c + leukocytes. Functionally, the leukocytes were dispensable for the initial migratory response of the regrowing endothelial sheet, but critical for the subsequent formation and maintenance of a front zone with high cellular density. Marker expression analyses, genetic fate mapping, phagocyte targeting experiments, and mouse knock-out experiments indicate that the CD11c + leukocytes were mononuclear phagocytes with an origin from both Ly6C high and Ly6C low monocytes. In conclusion, CD11c + mononuclear phagocytes are essential for a proper endothelial regrowth following arterial endoluminal scratch injury. Promoting the endothelial-preserving function of CD11c + leukocytes may be a strategy to enhance endothelial repair following surgical and endovascular procedures.

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Filter-Dense Multicolor Microscopy

Cited by 12 publications

References 25 publications

Site-specific programming of the host epithelial transcriptome by the gut microbiota

Site-specific programming of the host epithelial transcriptome by the gut microbiota

The regulation of gut mucosal IgA B-cell responses: recent developments

Endothelial repair is dependent on CD11c+ leukocytes to establish regrowing endothelial sheets with high cellular density

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