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. 2018 Sep 17;7(1):157–160. doi: 10.1016/j.jcmgh.2018.09.009

Figure 1.

Figure 1

Laser capture microdissection of mucus invaders. (A–F) Mucus layer staining on human colonic biopsy and LCM. (A) Muc2 (green). (B) Actin (red). (C) DNA/nucleus (blue, background owing to the use of PEN Membrane Frame Slides). (D) Selected area for microdissection. (E and F) After LCM, the inner mucus layer collected is transferred to the cap membrane and ready for DNA extraction. (G) 16S ribosomal RNA (rRNA) gene quantification after DNA extraction on unused cap membrane (negative control), and inner mucus microdissected from germ-free mice or E coli mono-associated mice. (H) Microbiota composition analysis of mucus invaders from E coli mono-associated mice. (I) Fecal and mucus-associated microbiota composition was analyzed by 16S rRNA gene Illumina sequencing. PCoA of the unweighted UniFrac distance is represented, with samples colored by sample type (feces and mucus). (J) Taxa summarization performed at the phylum level. Values are means ± SEM, N = 4–6. *P < .05 determined by a 1-way analysis of variance corrected for multiple comparisons with a Sidak post-test. AIEC, adherent-invasive E coli.