MicroscopyInnovations
passes through aqueous cell culture media and a tilted glass coverslip, and finally is collected by the detection objective. Special ZEISS proprietary optical elements in the detec- tion beam path compensate for these refractive index mis- matches. Alignment processes are performed automatically so that researchers can focus their full attention on their experiments. Te controlled incubation environment and an integrated auto-immersion mechanism enable unattended long-term experiments. Te high spatiotemporal resolution of the system allows for high-speed volumetric imaging of dynamics within cells with up to three volumes per second, as well as long-term observations of events like repeating mitosis over hours and days. Advantages over other lattice light-sheet microscopes include higher image resolution (compared to conventional Gaussian light sheet micros- copy), use of standard sample carriers, auto-alignment, and provisions for unattended experiments. Applications include live cell imaging of subcellular pro-
cesses with high speed (organelle dynamics, vesicle traffick- ing), 3-D imaging cell of migration within organoids, and 3-D imaging of small organisms with near isotropic resolu- tion (C. elegans, Drosophila). Lattice light sheet technology is acknowledged to be the most noninvasive fluorescence imaging method, allowing hundreds of 3-D image stacks, encompassing tens of thousands of images, to be taken from a single cell over extended time periods. Te Lattice Lightsheet 7 makes this technology available to a broad community.
METEOR for cryo-CLEM Delmic B.V.
Developers: Marit Smeets, Tomas van der Heijden, Wouter Roelofsen, Andries Efing, Éric Piel, and Bassim Lazem METEOR
is a fluorescence
light microscope (FLM) that can be integrated into a cryogenic focused ion beam (FIB)/scanning electron microscope (SEM) specimen cham- ber. METEOR enables in situ cryo- genic correlative light and electron microscopy (cryo-CLEM) to assist in producing lamella for cryo electron
tomography (cryo-ET). To image intracellular molecules at high resolution using a cryo-TEM, the part of the cell con- taining molecules of interest needs to be milled to a lamella 150–300 nm thick. Te milling is done using the FIB in a cryo- FIB/SEM. Since SEM provides only topographic informa- tion, fluorescence labeling and fluorescence microscopy are used to locate the molecules of interest. Before METEOR was introduced, researchers could only acquire cryo-FLM images of the sample using a separate cryo-FLM system outside the FIB. Specimen transfer between a cryo-FLM and a cryo-FIB/ SEM poses risks to the sample such as devitrification and ice
2021 September •
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contamination. Furthermore, researchers cannot check that they have captured the molecules of interest in the lamella without having a cryo-FLM integrated into the cryo-FIB/ SEM. Transfer of the prepared lamella to the cryo-TEM may be accomplished using the Delmic CERES Vitri-Lock to mini- mize devitrification and ice contamination during transfer. Te CERES Vitri-Lock is a high-vacuum and liquid-nitrogen- cooled version of the conventional “wobble stick.” METEOR is designed to be implemented in a FIB/SEM
through its existing ports without any permanent alterations to the instrument. Te METEOR light microscope optical path is placed parallel to the focused ion beam, allowing the user to switch seamlessly between FIB milling and FLM imag- ing. Te objective lens is attached to a PIEZO-driven stage, which enables precise FLM focusing and the acquisition of z-stacks. Te FLM optical path can be customized by select- ing the appropriate objective, dichroic mirror, and emission filters. Te user can navigate the sample and maintain focus as the imaging mode is switched among SEM, FIB, or FLM. Te incorporation of an FLM into the cryo-ET workflow
is especially important for rare or unknown biological cell structures. METEOR can be used for targeted milling of cells to study proteins important in disease pathways, for example, Parkinson’s disease, Huntington’s disease, and viral infections.
CLIMATE for in situ Microscopy DENSsolutions B.V.
Developers: Hugo Pérez-Garza, Ronald G. Spruit, Dan Zhou, Merijn Pen, and Christian Deen CLIMATE is a plug-and-play system
that relies on an innovative micro-elec- tromechanical system (MEMS) device, referred to as the Nano-Reactor, to accu- rately control the gas environment sur- rounding the specimen inside the TEM. Together with the Nano-Reactor, the CLIMATE system consists of a func- tionalized specimen holder, a heating control unit (HCU), a gas supply system (GSS), a vaporizer, and a residual gas analyzer (RGA). Te GSS controls the gaseous environment, allowing the user
to mix three different gases through mass flow controllers and a mixing valve. Te desired gas mixture is then sup- plied to the sample, where the user can control pressures and flow rates. Tese parameters can be changed in real time to achieve any gas mixture, at pressures from vacuum to 2000 mbar. Te system also offers the capability of controlling the humidity of the gas mixture from 0–100% relative humid- ity. Te HCU provides temperatures from room tempera- ture to 1200°C, which can be maintained with a stability of <0.01°C and settling times of a few seconds. Tese integrated
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