Billions of neurons are active in the human brain. They form flexible connections, which can be stabilized during information processing in learning and memory formation or destabilized in the reverse processes. The culture and imaging of neuronal cells is particularly challenging since mature neurons do not undergo cell division. One major problem associated with cultured neuronal networks is their short survival. Susceptibility to infection, hyperosmolality related to medium evaporation and the fact that it takes several weeks to months to observe network-wide activity makes increasing viability and extending neural life span in vitro extremely important. This type of stress leads to significant limitations for live cell imaging possibilities. The 3D Cell Explorer microscope would help avoid these perturbations and improve fundamental research on brain cells as the samples need no preparation, which allows for a fast, non-invasive and unbiased live cell observation. In addition, the 3D Cell Explorer laser uses 100 times less energy than the least energetic laser in the current fluorescent imaging approaches, which makes long-term imaging (up to weeks) possible.
Human Mesenchymal Stem Cell Differentiation into Neurons
To our knowledge this is the first high resolution long term live cell time-lapse movie imaging the differentiation of umbilical cord matrix human mesenchymal stem cells into neurons.
Cells were grown for 13 days in complete mesenchymal stem cell neurogenic differentiation medium before the differentiation process portrayed in this movie could be observed.
The cells in this footage were imaged for 20 hours every 30 seconds using Nanolive’s 3D Cell Explorer.
Cells and reagents were kindly provided by PromoCell.
Neural plasticity in live rat hippocampal neurons
In these videos we can observe rat hippocampal neurons in culture. The purple colored video is digitally stained and in 3D.
The cells were at 7 DIV, when they already become “mature neurons”. In this short video we can clearly observe two distinctive phenomena characterizing the normal physiology of this beautiful cell type: vesicle migration and neuronal plasticity.
A special thanks for the beautiful sample goes to the lab of prof. Šuput’s group at the Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
Mitochondrial trafficking in neurons
In this video, neuronal mitochondria were stained using a fluorescent dye and then imaged for 10 mins with the 3D Cell Explorer-fluo at an acquisition frequency of 1 image every 10 secs. Mitochondria can be seen being trafficked along axons, dendrites and within the cell body. They are also present in dendritic growth cones (actin-supported extensions of developing neurites).
Sample courtesy of Vanessa Morais and Rita Soares, iMM institute Lisbon.
Read our full blog post here: https://nanolive.ch/mitochondrial-trafficking-in-neurons/.
Neuronal Mitochondria Dynamics
In this spectacular video of live neuron cells we can observe the dynamics and behavior of living and completely unperturbed mitochondria moving inside axons, dendrites and around the nucleus.