Journal: Lab on a chip

Article Title: Multiplexed analysis of neural cytokine signaling by a novel neural cell-cell interaction microchip.

doi: 10.1039/d0lc00401d

Figure Lengend Snippet: Fig. 3 Immunocytology of neural rosettes formed on the NCCIM. (a) EB-derived rosettes in the dish and in the NCCIM. Cells are stained with DAPI, tight junction protein ZO1, NSC multipotency marker Nestin, neural ectoderm (Pax6), proliferation marker ki67, and neural progenitor (Sox1, Sox2) biomarkers as indicated. (b) Monolayer rosettes in the dish are dissociated and seeded into the NCCIM and form uniformly sized NSC rosette neurospheres by day 2. Cells are evaluated with similar biomarkers as (a). (c) Phase and fluorescence microscopy images of EB-derived rosettes in the PDMS microchambers of the NCCIM. Spontaneously formed rosettes in microwells results in three main size distribution categories: one large rosette, 3–4 medium size rosettes, and >5 small size rosettes. Cells are stained with ZO-1 (green) and Sox1 (red) antibodies. Scale bar is 50 μm.

Article Snippet: The antibodies, including Sox1 (R&D systems AF3360), Sox2 (R&D systems MAB2018), ZO-1 (ThermoFisher Scientific 33-9100), Ki67 (Abcam 15580), Nestin (R&D systems MAB1259), Pax6 (DSHB PAX6, Pax6 was deposited to the DSHB by Kawakami, A.) were added (1 : 1000) and incubated at 4 °C overnight.

Techniques: Derivative Assay, Staining, Marker, Fluorescence, Microscopy

Journal: Lab on a chip

Article Title: Multiplexed analysis of neural cytokine signaling by a novel neural cell-cell interaction microchip.

doi: 10.1039/d0lc00401d

Figure Lengend Snippet: Fig. 5 Viability and multipotency of rosettes on the NCCIM. (a) Flow diagram from seeding of cells into the PFMA that is combined with the MIST array to generate the assembled NCCIM. Cells were maintained within the NCCIM for 8 hours then evaluated by (b) propidium iodide (PI) staining of EB-derived rosettes and rosette neurospheres to visualize dead cells at hour 0 and hour 8. In (c) neural progenitor marker SOX1 is shown with adherens junction protein ZO-1 that highlights the center of polarized rosette cells. Cells maintain their multipotent state after NCCIM incubation for 8 hours. Scale bar is 50 μm.

Article Snippet: The antibodies, including Sox1 (R&D systems AF3360), Sox2 (R&D systems MAB2018), ZO-1 (ThermoFisher Scientific 33-9100), Ki67 (Abcam 15580), Nestin (R&D systems MAB1259), Pax6 (DSHB PAX6, Pax6 was deposited to the DSHB by Kawakami, A.) were added (1 : 1000) and incubated at 4 °C overnight.

Techniques: Staining, Derivative Assay, Marker, Incubation