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Hypotonic treatment optimization for enhanced intracellular visualization. (A) Optical micrographs of cells treated with three hypotonic solutions prepared by mixing distilled water (DW) and PBS—pure DW (0X, 100% DW), 0.2X PBS solution, and 0.5X PBS solution. Cells were imaged at 1, 5, 8 and 10 min post-treatment. White arrows indicate representative cells showing membrane rupture or cellular stress under stronger hypotonic conditions (0X and 0.2X PBS). Scale bar: 50 μm. (B) <t>Confocal</t> <t>microscopy</t> images of bEnd.3 cells after hypotonic treatment showing cell morphology with DAPI (blue) and cell mask staining. Left panel: 1X PBS control treatment for 10 min. Right panel: 0.5X PBS treatment for 10 min. Scale bar: 10 μm. (C) Quantitative analysis of cell height measured using NIS-E software program following 10 min treatment with 1X PBS (control) versus 0.5X PBS hypotonic solution. Data are presented as mean ± SEM ( n = 13).
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Hypotonic treatment optimization for enhanced intracellular visualization. (A) Optical micrographs of cells treated with three hypotonic solutions prepared by mixing distilled water (DW) and PBS—pure DW (0X, 100% DW), 0.2X PBS solution, and 0.5X PBS solution. Cells were imaged at 1, 5, 8 and 10 min post-treatment. White arrows indicate representative cells showing membrane rupture or cellular stress under stronger hypotonic conditions (0X and 0.2X PBS). Scale bar: 50 μm. (B) Confocal microscopy images of bEnd.3 cells after hypotonic treatment showing cell morphology with DAPI (blue) and cell mask staining. Left panel: 1X PBS control treatment for 10 min. Right panel: 0.5X PBS treatment for 10 min. Scale bar: 10 μm. (C) Quantitative analysis of cell height measured using NIS-E software program following 10 min treatment with 1X PBS (control) versus 0.5X PBS hypotonic solution. Data are presented as mean ± SEM ( n = 13).

Journal: Drug Delivery

Article Title: Cell swelling and upright mounting-based imaging for high-resolution visualization of intracellular trafficking across the BBB using conventional confocal microscopy

doi: 10.1080/10717544.2025.2608235

Figure Lengend Snippet: Hypotonic treatment optimization for enhanced intracellular visualization. (A) Optical micrographs of cells treated with three hypotonic solutions prepared by mixing distilled water (DW) and PBS—pure DW (0X, 100% DW), 0.2X PBS solution, and 0.5X PBS solution. Cells were imaged at 1, 5, 8 and 10 min post-treatment. White arrows indicate representative cells showing membrane rupture or cellular stress under stronger hypotonic conditions (0X and 0.2X PBS). Scale bar: 50 μm. (B) Confocal microscopy images of bEnd.3 cells after hypotonic treatment showing cell morphology with DAPI (blue) and cell mask staining. Left panel: 1X PBS control treatment for 10 min. Right panel: 0.5X PBS treatment for 10 min. Scale bar: 10 μm. (C) Quantitative analysis of cell height measured using NIS-E software program following 10 min treatment with 1X PBS (control) versus 0.5X PBS hypotonic solution. Data are presented as mean ± SEM ( n = 13).

Article Snippet: High-resolution apicobasal imaging was performed using LSCM (Nikon, A1Plus, Tokyo, Japan) equipped with a 60 × oil immersion objective lens (Apo 60 × oil λS DIC N2, numerical aperture = 1.40).

Techniques: Membrane, Confocal Microscopy, Staining, Control, Software

Confocal microscopy analysis of early endosome trafficking patterns. (A) Cells treated with A647-Tf for 5 min, 15 min, or 1 h, stained with DAPI (blue), Rab5 (green, upper panels), and EEA1 (green, lower panels). (B) Cells treated with A647-anti-TfR Ab under the same conditions and staining. The red signal corresponds to internalized A647-Tf or A647-anti-TfR Ab. Scale bar: 10 μm.

Journal: Drug Delivery

Article Title: Cell swelling and upright mounting-based imaging for high-resolution visualization of intracellular trafficking across the BBB using conventional confocal microscopy

doi: 10.1080/10717544.2025.2608235

Figure Lengend Snippet: Confocal microscopy analysis of early endosome trafficking patterns. (A) Cells treated with A647-Tf for 5 min, 15 min, or 1 h, stained with DAPI (blue), Rab5 (green, upper panels), and EEA1 (green, lower panels). (B) Cells treated with A647-anti-TfR Ab under the same conditions and staining. The red signal corresponds to internalized A647-Tf or A647-anti-TfR Ab. Scale bar: 10 μm.

Article Snippet: High-resolution apicobasal imaging was performed using LSCM (Nikon, A1Plus, Tokyo, Japan) equipped with a 60 × oil immersion objective lens (Apo 60 × oil λS DIC N2, numerical aperture = 1.40).

Techniques: Confocal Microscopy, Staining