Sphere formation ability of cells was assessed using spheroid assays according to the literature of Asadzadeh et al. [21 (link)]. Firstly, cell culture medium containing 10% Matrigel (354,234, Corning, Corning, New York, USA) was added into each well of the 96-well plates. Then, 2.5 × 103 cells were further added into each well and incubated for 10 days. Finally, the image of spheres in each well was observed under DMi8 S optical microscope (Leica) and captured at a magnification of × 200.
Dmi8 s optical microscope
Manufactured by Leica
The Leica DMi8 S is an optical microscope designed for laboratory use. It features a sturdy and ergonomic design, and is equipped with high-quality optics to provide clear and detailed images. The microscope's core function is to magnify and observe samples under high magnification for scientific and research applications.
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Lab products found in correlation
2 protocols using dmi8 s optical microscope
1
Spheroid Formation Assay for Cell Analysis
2
Coculture of Neurons and Astrocytes for In Vitro Modeling
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A major advantage of the coculture system over neurons alone is to support the growth, survival, and differentiation of nutrient factors secreted by the glial feeding layer, which more accurately resembles the environment in vivo; moreover, the coculture can be applied to explore neuronal-glial interactions (36 ). The coculture of neuron with astrocyte was performed as previously illuminated (17 (link), 37 (link)). Before coculture, transfected or untransfected astrocytes were cultured in 24-well plates. Meanwhile, neurons (1 × 104 cells/coverslip) were plated in glass coverslips to which 3 to 4 beads of paraffin (A13912, OKA) were previously affixed. After 30 min (min) of incubation in neuronal culture medium to allow for neurons to attach, the glass coverslips were inverted in a 24-well plate above the astrocyte monolayer (the paraffin drops to prevent neuron-astrocyte contact) for coculture for 3 days. After that, the glass coverslips were taken out from the 24-well plate and the neurons on the glass coverslips were firstly observed under a DMi8 S optical microscope (Leica) at 400× magnification. Then, the growth and viability of neurons were detected by microtubule-associated protein 2 (MAP2) immunofluorescence staining and methyl thiazolyl tetrazolium (MTT) assay, respectively.
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