Ten-thousand LN229-TS cells were seeded into one well of a 24-well plate, followed by addition of NP-DiI (2 μl) 12 h later. Cells were rinsed with PBS twice (500 μl each) followed by fixation with 4% PFA at 30 min and 6h after addition of NP-DiI. Afterwards, the cells were rinsed with PBS three times, incubated in blocking buffer (1XPBS, 5% BSA, 0.3% Triton X-100) at room temperature for 1 h followed by three washes with PBS and staining with an EEA1 antibody (CST, cat. no. 3288S, 1:200 in Antibody Dilution Buffer (1X PBS / 1% BSA / 0.3% Triton X-100) at 4 °C for 24 h. Afterwards, the cells were washed with PBS three times (5 min each), incubated with secondary antibody at room temperature for 1.5 h and washed with PBS three times (5 min each). Prolong Gold Antifade Reagent with DAPI (CST, cat. no. 8961, 5 μl) was utilized to mount the cells. Images of cells were collected using a confocal microscope (Olympus, x100 oil objective).
X100 oil objective
Manufactured by Olympus
The X100 oil objective is a high-quality microscope objective designed for use with oil immersion techniques. It provides a magnification of 100x and a numerical aperture of 1.25, enabling high-resolution imaging and detailed observation of samples.
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2 protocols using x100 oil objective
1
Cellular Uptake and Localization of NP-DiI
2
Cellular Uptake and Localization of NP-DiI
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Ten-thousand LN229-TS cells were seeded into one well of a 24-well plate, followed by addition of NP-DiI (2 μl) 12 h later. Cells were rinsed with PBS twice (500 μl each) followed by fixation with 4% PFA at 30 min and 6h after addition of NP-DiI. Afterwards, the cells were rinsed with PBS three times, incubated in blocking buffer (1XPBS, 5% BSA, 0.3% Triton X-100) at room temperature for 1 h followed by three washes with PBS and staining with an EEA1 antibody (CST, cat. no. 3288S, 1:200 in Antibody Dilution Buffer (1X PBS / 1% BSA / 0.3% Triton X-100) at 4 °C for 24 h. Afterwards, the cells were washed with PBS three times (5 min each), incubated with secondary antibody at room temperature for 1.5 h and washed with PBS three times (5 min each). Prolong Gold Antifade Reagent with DAPI (CST, cat. no. 8961, 5 μl) was utilized to mount the cells. Images of cells were collected using a confocal microscope (Olympus, x100 oil objective).
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