Imaging Biological Samples – a Reference List
In Light Sheet Fluorescence Microscopy (LSFM), the detection beam path is placed perpendicular to the illumination beam path. By these means a fluorescently labeled sample is illuminated from the side, using laser light that is formed into a thin sheet of light, exciting only fluorophores within the focal plane of the detection objective. Thus all fluorescent signal can be collected on a camera-based detector. This unique setup allows for extremely light efficient optical sectioning and unprecedented imaging speed, causing only minimal phototoxicity or bleaching. Image acquisition with high spatial and temporal resolution over long periods of time becomes a feasible task.
ZEISS Lightsheet Z.1 offers a horizontal LSFM setup, in which a sample is suspended from above and placed into a liquid-filled sample chamber, providing the sample with environmentally (temperature, medium) stable conditions over long periods of time. Additionally, the sample can be rotated in front of the detection objective to image from the perfect angle or to combine different viewing perspectives into one dataset, known as Multiview imaging. The many advantages of Lightsheet Z.1 have been used by scientists of different specialties to advance their research. Imaging live specimens clearly benefits from the combination of advantages LSFM offers. Therefore, many publications featuring Lightsheet Z.1 are reporting in vivo imaging of whole organisms (e.g. model organism in developmental biology) or three-dimensional cell and tissue cultures. Even fixed and chemically cleared tissues, such as whole mouse brains, also make use of the fast, sensitive and stable imaging conditions of Lightsheet Z.1.
A new Application Note details the published research using ZEISS Lightsheet Z.1 over the past three years. This reference list shows the vast opportunities of a multi-purpose LSFM within different areas of research. These include scientific publications in cell culture and in vitro imaging, clearing applications, plant imaging, whole organism in vivo imaging, and image processing.
- Olaf Selchow and Jan Huisken (2013). Light sheet fluorescence microscopy and revolutionary 3D analyses of live specimens. Photonik international, 2013
- Reynaud, E. G. et al (2015). Guide to light-sheet microscopy for adventurous biologists. Nature Methods 12, 30 – 34. doi:10.1038/nmeth.3222
- Pampaloni, F. et al (2015). Light sheet-based fluorescence microscopy (LSFM) for the quantitative imaging of cells and tissues. Cell and tissue Research Volume 360, Issue 1 (129 – 141). doi 10.1007/s00441-015-2144-5
- White Paper: Sample Preparation for Light Sheet Microscopy, Protocols and Guidelines for ZEISS Lightsheet Z.1