MICROSCOPY & IMAGING
asymmetric configuration of the illumination and detection objectives (10x/0.3NA and 25x/1.1NA, respectively). Simultaneous two-colour imaging is enabled by spectrally separating two different channels on two sCMOS cameras, both of which can acquire more than 80fps at full frame. Suited for cell culture applications, this microscope design features a small U-shaped sample compartment (Fig. 2A). Tis is covered in FEP foil (optically transparent material), which serves as a physical barrier between the sample medium and the immersion medium (Fig. 2B). Te foil works as a “curved coverglass” where cells can be cultured or spheroids and small embryos can be placed. Te temperature, humidity, CO2
can be controlled, allowing for a variety of conditions to be established. Tis system is suited to a variety of samples and applications,
and O2
including: in toto imaging of small animal and embryo models, observing dynamic processes in mammalian cell culture models (e.g. spheroids, organoids), live imaging of plant models, investigating stem cell development and differentiation, in vitro fertilisation research and monitoring, and functional imaging (calcium). Another recent innovation in design (InVi SPIM Lattice Pro) offers tailorable, interactive adaptability of the beam shape to suit the specific requirements of a sample. Tis provides flexibility for the type of light-sheets that can be generated, e.g. single or multiple variable Bessel beams, lattice light-sheets for structured illumination, and static or scanned Gaussian beams (Fig. 3A). It enables researchers to combine different types of beams in one experiment and select the setting that gives the best results for their specific 3D high-resolution imaging experiments: large field of view, high speed and optimal spatial resolution (Fig. 3B).
Te range of applications includes observing dynamic cellular interactions, cell cycle imaging, studying membrane dynamics, subcellular structure visualisation and long time-lapse imaging. By only exciting the fluorophores in a single plane, light-sheet
microscopy is gentle enough to image specimens over extended periods. Additionally, the technique’s high imaging speed allows researchers to follow the rapid development of organisms not easily observed with traditional scanning microscopes.
levels of the incubation chamber
Nanopositioning for Microscopy
FAST, COMPACT, TO THE NANOMETER
Fig.3. Flexible light-sheet illumination patterns enable combining different types of beams in one experiment. (A) Optimal imaging results (marked with a blue box) were obtained with the optical lattice (EGFP labelling) and the Gaussian beam (mRFP labelling). (B) Overlay image. Courtesy of Dr. Martin Stöckl, University of Konstanz, Germany. Images taken with the InVi SPIM Lattice Pro.
Carolina Araya is with Luxendo, a Bruker company.
www.luxendo.eu
PI – The Broadest and Deepest Portfolio in Precision Motion Technologies.
Physik Instrumente (PI) GmbH & Co. KG
www.pi.ws ·
info@pi.ws · +49 721 4846-0
MOTION | POSITIONING
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