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Axio imager 2

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The Axio Imager 2 is a high-performance microscope system designed for advanced scientific applications. It features a modular design, allowing for customization to meet the specific needs of researchers and professionals. The microscope provides superior optical performance, enabling detailed observation and analysis of samples.

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Lab products found in correlation

Dm5000 b, by Zeiss (2 mentions) Whatman, by Cytiva (1 mentions)

3 protocols using axio imager 2

1

Tissue Fixation and Sectioning for Immunofluorescence and In Situ Hybridization

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For tissue staining in section, mice were perfused with cold 4% PFA in PBS. The proximal 10 cm of duodenum was cleaned, flushed with cold 4% PFA, and fixed in 4% PFA for 4 hours at 4°C. The tissue was cryo-protected in 30% sucrose overnight at 4°C. Samples were embedded in OCT and 8 µm sections were prepared for immunofluorescence staining. For immunohistochemistry and in situ hybridization, the samples were fixed overnight in 4% PFA, paraffin embedded, and sectioned at 5 µm for immunohistochemistry or 10 µm for in situ hybridization. For crypt area quantitation, crypts clearly above distended granuloma tissue containing visible larval worms were called as “gran” and others were called “non-gran”. Crypt area was quantitated in ImageJ. For fetal whole mount imaging, fetal intestines were fixed in 4% PFA in PBS for 3 hrs, permeabilized, and blocked for 4 hours at room temperature. Primary and secondary antibodies were incubated at 4°C overnight. Images were acquired and processed with a Leica DM5000 B or a Zeiss Axio Imager 2 and Adobe Photoshop.
Nusse Y.M., Savage A.K., Marangoni P., Rosendahl-Huber A.K., Landman T.A., de Sauvage F.J., Locksley R.M, & Klein O.D. (2018). Parasitic helminthes induce fetal-like reversion in the intestinal stem cell niche. Nature, 559(7712), 109-113.
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2

Intestinal Tissue Preparation for Immunofluorescence

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For analysis by immunofluorescence, tissues were harvested, flushed extensively with cold PBS, and fileted. For whole mount, fileted intestine was washed with DTT/EDTA and rinsed in HBSS, as for flow cytometry above. Intestinal pieces were laid flat between Whatman paper with light pressure for 4 hours in 2% paraformaldehyde with 2 mM MgCl2 and 1.25 mM EGTA at 4°C, then washed overnight in excess PBS. For thin section, the tissue was also dehydrated in 30% sucrose for >4 hours before rolling the tissue on a wooden stick, mounting in OCT as a ‘swiss roll’, and cutting 8 um sections with a cryostat. Thin sections were stained and mounted per standard procedures. Tissue for whole mount was stained in 0.5% TritonX-100 per standard procedures. Images were acquired and processed with a Zeiss Axio Imager 2 and Adobe Photoshop.
Savage A.K., Liang H.E, & Locksley R.M. (2017). The development of steady-state activation hubs between adult LTi ILC3s and primed macrophages in small intestine. Journal of immunology (Baltimore, Md. : 1950), 199(5), 1912-1922.
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3

Tissue Fixation and Sectioning for Immunofluorescence and In Situ Hybridization

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For tissue staining in section, mice were perfused with cold 4% PFA in PBS. The proximal 10 cm of duodenum was cleaned, flushed with cold 4% PFA, and fixed in 4% PFA for 4 hours at 4°C. The tissue was cryo-protected in 30% sucrose overnight at 4°C. Samples were embedded in OCT and 8 µm sections were prepared for immunofluorescence staining. For immunohistochemistry and in situ hybridization, the samples were fixed overnight in 4% PFA, paraffin embedded, and sectioned at 5 µm for immunohistochemistry or 10 µm for in situ hybridization. For crypt area quantitation, crypts clearly above distended granuloma tissue containing visible larval worms were called as “gran” and others were called “non-gran”. Crypt area was quantitated in ImageJ. For fetal whole mount imaging, fetal intestines were fixed in 4% PFA in PBS for 3 hrs, permeabilized, and blocked for 4 hours at room temperature. Primary and secondary antibodies were incubated at 4°C overnight. Images were acquired and processed with a Leica DM5000 B or a Zeiss Axio Imager 2 and Adobe Photoshop.
Nusse Y.M., Savage A.K., Marangoni P., Rosendahl-Huber A.K., Landman T.A., de Sauvage F.J., Locksley R.M, & Klein O.D. (2018). Parasitic helminthes induce fetal-like reversion in the intestinal stem cell niche. Nature, 559(7712), 109-113.
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