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dapi signal  (Oxford Instruments)
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; <t>DAPI</t> max pojection of isolated villi for dotted region in leftmost image; Imaris <t>based</t> <t>3D</t> projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).
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Image Search Results


(A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; DAPI max pojection of isolated villi for dotted region in leftmost image; Imaris based 3D projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).

Journal: bioRxiv

Article Title: Ontogeny of the vagal gut-brain axis

doi: 10.1101/2025.05.01.651736

Figure Lengend Snippet: (A) Wholemount fluorescence microscopy of the embryonic day 16.5 (E16.5) intestine. Left to right: single focal plane of proximal intestine; max z-projection of the same field; DAPI max pojection of isolated villi for dotted region in leftmost image; Imaris based 3D projections of villi. (B) Quantification of the frequency of Nd1+ cells per villus from E15.5 and E16.5 proximal intestines. Small dots at left represent single villus; large dots at right represent average per embryo. Data presented are from 4 individuals and at least 2 separate litters. (C) Quantification of the frequency of CCK+ cells per villus from E15.5 and E16.5 proximal intestines, shown as in (B). (D) Max projections of mGFP (black) proximal intestines from Nd1 mGFP embryos at E15.5 (left) or E16.5 (right). (E) Nd1+ cell (EEC) density based on embryonic age and intestinal region. Each dot represent data from one individual embryo, with at least 2 separate litters represented per timepoint. (F) Wholemount E16.5 proximal intestine stained for Nd1 mGFP (green), substance P (SubP; magenta) and cholecystokinin (CCK; yellow) showing max projection (left) and single plane images of individual EECs (right). (G) Quantification of EEC subtype based on accumulation of CCK and/or SubP within Nd1 mGFP + epithelia; n indicates individual embryos from at least two separate litters per timepoint. (H,I) Max projection images of proximal intestine at E16.5 (H) or E15.5 (I) stained for Tuj1. Right-most panels depict individual villi outlined in white dashed boxed in the first two panels. (J) Max z-projection image of E15.5 myenteric plexus from proximal intestine stained for Tuj1 (magenta) to label all axons. Scale bars, 100 m (A, first three panels), 25 µm (A, right), 40 µm (D,F), 75 µm (H-J).

Article Snippet: To determine sample volume, we generated a 3D surface of the DAPI signal in Imaris, using a minimal threshold to generate the surface in a manner reflecting the tissue shape.

Techniques: Fluorescence, Microscopy, Isolation, Staining