proteome profiler human xl cytokine array kit  (R&D Systems)

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: Illustration of MACSima™ Imaging Cyclic Staining (MICS) principle MICS technology was applied (Step 46). (0) Image acquisition of 3D-IF staining in autofluorescence channel, followed by Photobleaching. (2–4) Multi-cyclic imaging: Rounds of 2D-IF staining with FITC, PE and APC coupled antibody fluorochrome-conjugate, image acquisition of respective FITC, PE and APC-channels and signal erasure by photobleaching.

Article Snippet: MACSima Stain Support Kit, mouse/human , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-575/Cat# 130-127-574.

Techniques: Imaging, Staining

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: MACSima Stain Support Kit, mouse/human , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-575/Cat# 130-127-574.

Techniques: Imaging, Comparison, Selection, Staining

Journal: STAR Protocols

Article Title: Protocol to generate human stem cell-derived CD70-directed allogeneic CAR-NKT cells for treating renal cell carcinoma

doi: 10.1016/j.xpro.2025.104340

Figure Lengend Snippet: Generation of Allo CAR70-NKT cells from HSPCs using a clinically guided culture method (A and B) Schematics showing the generation of Allo CAR70-NKT cells (A) and the design of Lenti/iNKT-CAR70-IL-15 lentivector (B). (C) FACS detection of NKT TCR expression in lentivector-transduced CD34 + HSPCs. (D) Quantification of (C) (n = 5; n indicates different cord blood donors). (E) FACS monitoring of the generation of Allo CAR70-NKT cells during the 6-week culture. (F) Percentage of Allo CAR70-NKT cells in total live cells during the 6-week culture (n = 5; n indicates different cord blood donors). (G) Yield of Allo CAR70-NKT cells (n = 5; n indicates different cord blood donors). (H) CAR70 expression on Allo CAR70-NKT cells (n = 5; n indicates different cord blood donors). (I) ELISA measurements of IL-15 production by Allo CAR70-NKT cells with or without αGC stimulation (n = 4). (J) CD4/CD8 subpopulations of Allo CAR70-NKT cells. Data generated from 5 cord blood donors are shown. SP, single-positive; DP, double-positive; DN, double-negative. (K–M) Antigen responses of Allo CAR70-NKT cells. Allo CAR70-NKT cells were stimulated with/without αGC-loaded PBMCs for 1 week. (K) Experimental design. (L) Growth curve of Allo CAR70-NKT cells (n = 4). (M) ELISA measurements of effector cytokine levels in the culture supernatants collected on day 5 (n = 4). Representative of over 5 experiments. Data are presented as the mean ± SEM. ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 by Student’s t test (I and M).

Article Snippet: Human CD34 MicroBeads Kit , Miltenyi Biotec , CAT#130-046-703.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Generated