DetailsAir can be used to image tissue. For dried cells, imaging is possible but morphological details will be significantly compromised and this environment is not compatible with live cell imaging. A disadvantage of air and dry samples is that it can easily give rise to heating of the sample since there is poor hear transfer from the sampling volume.
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DetailsWhile water is compatible with Raman imaging, the difference in salt concentration between the cell cytoplasm and the water can cause cytolysis - bursting of the cell due to osmotic imbalance. If imaging hydrated cells, DPBS should be used instead where possible.
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DetailsDPBS, Dubeccos' Phosphate-Buffered Saline is the ideal environment for fixed-cell imaging as it maintains the osmotic balance of cells and has minimal Raman signal.
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DetailsCell media is ideal for live imaging applications, though it is important to use phenol red-free media due to the fluorescence contribution of phenol red. In addition, if imaging in a non-CO2 controlled environment, a CO2-independent medium such as Lebovitz L-15 medium should be used.
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Details3D hydrogels provide an environment that most accurately reconstructs the in vivo environment. While cell imaging in hydrogels is possible, it can be challenging to identify regions of interests using widefield imaging. In addition, choice of matrix material needs to be considered in terms of its autofluorescence and gel Raman signal.
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