Journal: Frontiers in Immunology

Article Title: Altered intraperitoneal immune microenvironment in patients with peritoneal metastases from gastric cancer

doi: 10.3389/fimmu.2022.969468

Figure Lengend Snippet: Gating strategy of lymphocytes, myeloid cells and tumor cells in peritoneal fluid. An example of gating method analyzed by Flow Jo software. Among the DAPI(+) peritoneal free cells, dead cells were excluded with FVS780. Tumor cells and leukocytes were defined as CD45(-)CD326 (+) and CD45(+)CD326(-) cells, respectively, and tumor leukocyte ratio (TLR) was calculated as described in Material and Methods. Among the leukocyte populations, lymphocytes and myeloid cells were distinguished as FSC/SCC profile, and expression of antigens was further examined in each cell population.

Article Snippet: APC-conjugated mAb to CD326 (EpCAM) was purchased from Miltenyi Biotec (Auburn, CA).

Techniques: Software, Expressing

Journal: STAR Protocols

Article Title: Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing

doi: 10.1016/j.xpro.2025.104296

Figure Lengend Snippet: 3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Article Snippet: CD326 (EpCam) antibody, anti-human, Vio 780, REAfinity , Miltenyi Biotec B.V. & Co. KG , Cat# 130-136-631 RRID: AB_2904949.

Techniques: Imaging, Comparison, Selection, Staining

Journal: PLoS Pathogens

Article Title: Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment

doi: 10.1371/journal.ppat.1005641

Figure Lengend Snippet: ( A ) Cell entry is EpCAM dependent but independent of ephrin-B2. Representative flow cytometry plots out of three independent experiments of CHO-K1, CHO-EpCAM, CHO-ephrin-B2 cells, and of a mixed culture composed of CHO-EpCAM and CHO-ephrin-B2 (1:1 ratio) monitored 72 h after transduction with NiVmut EpCAM -LV, NiVwt-LV or VSV-LV (MOI of 1). EpCAM expression was detected by an APC-coupled human EpCAM specific antibody. ( B ) To ascertain stability of transduction with the EpCAM-targeted vector, CHO-EpCAM cells were cultivated for further 30 days after transduction with the indicated MOIs. The percentage of GFP-positive cells was determined by flow cytometry at the indicated time points. One representative out of three independent experiments is shown.

Article Snippet: Human EpCAM was detected by an Allophycocyanin (APC) labeled mouse anti-EpCAM antibody (clone HEA-125, Miltenyi Biotec, Bergisch Gladbach, Germany, dilution 1:100).

Techniques: Flow Cytometry, Transduction, Expressing, Plasmid Preparation