eclipse ts2 confocal microscope  (Carl Zeiss)

Journal: Biology Open

Article Title: Primary human neutrophils and monocytes migrate along endothelial cell boundaries to optimize search efficiency under static in vitro conditions

doi: 10.1242/bio.061704

Figure Lengend Snippet: Primary human monocytes cultured on top of monolayer of HUVECs migrate along cell–cell boundaries. (A) Primary human monocytes (Hoechst, red) enrich at cell–cell boundaries of HUVECs (α-Catenin, white). To the right, quantification of cells located at cell–cell boundaries (green) and on top of the cell (magenta) is shown ( N =3, n =196 cells). (B) Immuno-cytochemistry indicates that primary human monocytes transmigrate the HUVEC layer. Both the nuclei of monocytes (small) and HUVECs (large) are stained with Hoechst. (C) Statistical analysis of motion pattern for primary human monocytes and HUVECs. At the top, representative tracks are shown ( N =3, n =12 technical repeats). The dashed lines serve as a guidance to the eye. At the bottom, to the left, graph depicting the number of cells, normalized to the initial count, throughout the entire acquisition period. To the right, graph tracking speed over time. Each blue line represents a technical repeat. (D) Primary human monocytes (Hoechst, red) migrate along cell–cell boundaries (actin, white) of HUVECs. Only the nucleus of monocytes is labelled by Hoechst. (E) Motion tracks of human monocytes on top of HUVECs. At the top, Voronoi of endothelial layer (gray) as well as tracks of monocytes and of HUVECs are shown. Insets to the right are 70 µm×70 µm. Below, to the left, cross-correlation analysis shows enrichment of monocyte tracks along endothelial cell–cell boundaries identified by Vornoi tesselation. As negative control, to the bottom right, one channel was rotated by 180° prior to cross-correlation analysis. (F) Time-lapse of primary human monocytes (red) show different migration pattern. Both, the nuclei of monocytes (small) and HUVECs (large) are stained with Hoechst. (G) Primary human monocytes change fluorescence intensity of nucleus during transmigration. Both, the nuclei of monocytes (small) and HUVECs (large) are stained with Hoechst. (H) ER-HoxB8 derived monocytes/macrophages migrate along HUVEC boundaries. To the left, HUVECs (green) and tracks of ER-HoxB8 derived monocytes/macrophages (magenta) are shown. To the right, scanning electron microscope show ER-HoxB8 derived monocytes/macrophages at cell–cell boundaries of a confluent HUVEC sheet. Scale bars: (A,D,F,G) 20 µm, (B) 10 µm, (C,E) 100 µm, (H) 200 µm and 500 µm.

Article Snippet: Cell migration was imaged using an inverted confocal microscope (Nikon, Eclipse Ts2) with a digital camera (Nikon DS-Fi2), using a 20× objective at 37°C and 5% CO 2 with a frame interval of 60 s. Using multi-frame imaging allowed the recording of multiple positions per channel (i.e. technical repeats).

Techniques: Cell Culture, Immunocytochemistry, Staining, Negative Control, Migration, Fluorescence, Transmigration Assay, Derivative Assay, Microscopy

Journal: Redox Report : Communications in Free Radical Research

Article Title: Neferine improves oxidative stress and apoptosis in benign prostate hyperplasia via Nrf2-ARE pathway.

doi: 10.1080/13510002.2021.1871814

Figure Lengend Snippet: Protective effect of NF against OS: Human BPH-1 cells were treated with NF 3, 9 µM and SFN 15 µM for 48 h . A. ROS was determined with DCFH-DA staining and observed under Laser scanning confocal microscope (x 100) B. DCFH-DA fluorescence intensity representation. C. Determination of SOD, GST and MDA activity at 48 h. Values are presented as means ± SD (n = 5). For NF: * P < 0.05, ** P < 0.01 compared to control group. For SFN: # P < 0.05, ## P < 0.01 compared to that of the control group (no treatment).

Article Snippet: Subsequently, the cells are incubated in a 4′, 6-diamidino-2-phenylindole (DAPI) solution for 3 min, and the images were acquired using a Laser Scanning Confocal Microscope (Nikon Eclipse Ts2, Japan).

Techniques: Staining, Microscopy, Fluorescence, Activity Assay, Control