Endosomes were imaged using a 100x 1.4 NA oil DIC Plan-Apochromat VC objective (Nikon) with a Nikon A1 scanning confocal microscope. Photobleaching were performed in circular regions with 4 iterations of 488nm at 100% laser intensity. For the analysis, the fluorescence background was subtracted from ROIs intensity values. Those values were subsequently normalized by a non-bleached area and the value of intensity of the first frame after bleaching was subtracted. Finally, FRAP curves were normalized with the pre-bleach fluorescence intensity value. To determine FRAP recovery kinetics, we fitted with the following double exponential function f(t)=α(1-et-t0/τ1)+(1-α)(1-et-t0/τ2).
1.4 na oil dic plan apochromat vc objective
Manufactured by Nikon
The 100x 1.4 NA oil DIC Plan-Apochromat VC objective is a high-performance microscope objective designed for use with oil immersion techniques. It features a numerical aperture of 1.4 and a magnification of 100x, making it suitable for high-resolution imaging applications. The objective utilizes a Plan-Apochromat optical design and DIC (Differential Interference Contrast) capabilities to provide superior image quality and contrast.
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2 protocols using 1.4 na oil dic plan apochromat vc objective
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Measuring Endosome Dynamics by FRAP
2
Measuring Endosome Dynamics by FRAP
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Endosomes were imaged using a 100x 1.4 NA oil DIC Plan-Apochromat VC objective (Nikon) with a Nikon A1 scanning confocal microscope. Photobleaching were performed in circular regions with 4 iterations of 488nm at 100% laser intensity. For the analysis, the fluorescence background was subtracted from ROIs intensity values. Those values were subsequently normalized by a non-bleached area and the value of intensity of the first frame after bleaching was subtracted. Finally, FRAP curves were normalized with the pre-bleach fluorescence intensity value. To determine FRAP recovery kinetics, we fitted with the following double exponential function f(t)=α(1-et-t0/τ1)+(1-α)(1-et-t0/τ2).
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