To determine cell diameters, cells were detached with trypsin–EDTA (0.05%), trypsinisation was stopped by addition of appropriate cell culture media. Afterwards the media was replaced with PBS and bright-field imaging was performed using Nikon Eclipse Ti (Nikon) equipped with 20×, 0.45 numerical aperture Plan Fluor air objective (Nikon). The cell diameter was determined using Fiji software.51 (link)
Plan fluor air objective
Manufactured by Nikon
The Plan Fluor air objective is a high-quality optical lens designed for use in microscopy applications. It provides a flat, distortion-free image field and is optimized for use with air-based specimens. This objective is suitable for a variety of imaging techniques, including brightfield and fluorescence microscopy.
Automatically generated - may contain errors
2 protocols using plan fluor air objective
1
Determining Cell Diameters by Imaging
2
Characterization of Micro-Particles and Encapsulation
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The dimensions of the rough-smooth dumbbells
and spherical particles were determined by analyzing TEM images taken
using a FEI Tecnai 10 transmission electron microscope. The size distribution
of the microspheres was obtained from the analysis of optical microscopy
images using the program ImageJ.12 ,13 (link) The surface roughness of the microspheres and dumbbells was investigated
using a scanning electron microscope (SEM XL FEG 30, Philips). A Malvern
ZetaSizer Nano-ZS was used to measure both the polymer size of the
depletant with dynamic light scattering (DLS) and the zeta potential
of both the microspheres and dumbbells using laser doppler electrophoresis.
Encapsulation of the microspheres with dumbbell particles was studied
using a Nikon Eclipse Ti optical microscope with a Nikon Plan Fluor
air objective (NA = 0.75, 40× magnification). A Nikon Apo TIRF
oil immersion objective (NA = 1.49, 100× magnification) was used
to study the encapsulation of microspheres by the smaller spherical
particles. All images were acquired with an additional 1.5× magnification.
Image acquisition was performed using a Hamamatsu Digital Camera ORCA-Flash4.0
C11440 and the NIS-Elements Imaging Software.
and spherical particles were determined by analyzing TEM images taken
using a FEI Tecnai 10 transmission electron microscope. The size distribution
of the microspheres was obtained from the analysis of optical microscopy
images using the program ImageJ.12 ,13 (link) The surface roughness of the microspheres and dumbbells was investigated
using a scanning electron microscope (SEM XL FEG 30, Philips). A Malvern
ZetaSizer Nano-ZS was used to measure both the polymer size of the
depletant with dynamic light scattering (DLS) and the zeta potential
of both the microspheres and dumbbells using laser doppler electrophoresis.
Encapsulation of the microspheres with dumbbell particles was studied
using a Nikon Eclipse Ti optical microscope with a Nikon Plan Fluor
air objective (NA = 0.75, 40× magnification). A Nikon Apo TIRF
oil immersion objective (NA = 1.49, 100× magnification) was used
to study the encapsulation of microspheres by the smaller spherical
particles. All images were acquired with an additional 1.5× magnification.
Image acquisition was performed using a Hamamatsu Digital Camera ORCA-Flash4.0
C11440 and the NIS-Elements Imaging Software.
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