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Cells were incubated at 37°C overnight, washed in PBS and split into five different 6.4-mm wells in a 96-well flat bottom suspension plate.
For the sphere cell viability, the spheres were formed in sphere-forming conditions and enzymatically dissociated cells were cultured in the suspension plate, and then cells were treated with indicated agents.
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Extracellular bacteria suspensions and intracellular suspension plates were incubated at 37 °C overnight.
The specific design goal is to find the shape and orientation angle of the suspension plates that satisfy the desired fundamental eigenfrequencies in tracking, focusing and rolling directions.
In order to satisfy the stringent specification for the dynamic characteristics of modern optical pickup actuators, the topology design optimization of their suspension plates is presented.
The topology optimization considering the orientation angle yielded optimal suspension plates satisfying the target eigenfrequencies, which are not possible to obtain without considering it.
Digested tissue was finely disaggregated with Pasteur pipettes and single cell suspension plated and propagated in DMEM medium (Invitrogen) with 10% FBS (Sigma).
The NTERA2 stem cells were mixed at a ratio of one NTERA2.Tom cell per 2000 wildtype cells and the cell suspension plated at a density amenable to neuronal differentiation (15000 cells/cm2), in the presence of 10−5M RA.
During all the differentiation protocol, the cells were kept in suspension plates.
Briefly, tissue specimens were digested by type I collagenase (EC 3.4.24.3 – clostridiopeptidase A, Sigma-Aldrich, St . LouisMO, USA – Milan, Italy) and the suspension plated.
The fluid spheroid-containing gel was transferred into prewarmed 24-well suspension plates (Greiner Bio-One) and incubated for 30 min at 37°C.
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