MicroscopyInnovations
clinical examination of specimens under traditional light microscopes.
Delphi Phenom-World BV
Developers: Pieter Kruit, Andries Ef ing, and Sander den Hoedt
Delphi integrates fl uorescence microscopy (FM) and electron microscopy (EM) in a single device by equipping a tabletop scanning electron microscope (SEM) with an inverted fl uorescence microscope. T is
integration enables scientists to correlate two diff erent types of information on the same cell, tissue, or structure of interest. Although correlative light electron microscopy (CLEM) is becoming more popular, the complexity of the workfl ow has limited its widespread adoption. Delphi makes correlative microscopy more accessible by its compact design and user friendliness. T e inverted fl uorescence microscope in the Delphi is placed underneath the electron beam and allows researchers to seamlessly switch between the two imaging modalities. T is setup eliminates the need to search for a region of interest (ROI) because the light optical beam and the electron beam are always aligned on the ROI. T e integrated design of the Delphi, together with its open-source soſt ware package, make the Delphi easy to operate. T e intuitive soſt ware package allows the user to control all light microscopy settings as well as all SEM settings. T e Delphi comes with a sample holder designed to carry a
14 × 14 mm coverslip. T e sample holder has carbon tape corners enabling the user to locate the coverslip in the correct position. T e sample is open to the electron beam and transparent to the light beam.
Along with simplifying correlative microscopy, the Delphi reduces correlative workflow time. Sample loading time for the Delphi is about 2 minutes, and, after loading, the user can immediately begin correlative imaging. Precision mechanisms and the use of a proprietary automated alignment technique ensure precise, user-independent overlay of FM and EM images using cathodoluminescence, which is generated as the electron beam hits the glass coverslip.
Application areas are quite broad. T e main uses of the Delphi, however, are found in the life sciences where correl- ative microscopy is getting attention for its ability to combine structural and functional information. For whole cells, Delphi permits the study of cell surface morphology in correlation with specifi c proteins of interest. Using the Delphi for thin sections allows one to add fl uorescent markers that can pinpoint regions of interest, screen large areas, and identify subcellular structures on a molecular basis.
2015 September •
www.microscopy-today.com
MultiSEM 505 Carl Zeiss Microscopy GmbH
Developers: Dirk Zeidler, Sebastian Vollmar, Arne T oma, Robert Schwede, Dieter Schumacher, Stefan Schubert, Karin Schiele, Christof Riedesel, Nicole Rauwolf, Stephan Nickell, Mario Mützel, Ingo Müller, Ralf Lenke, Dieter Lechner, Uwe Lang, Jürgen Kynast, Paul Kortyka, T omas Kemen, Nicolas Kaufmann, Nico Kämmer, Jörg Jacobi, Tomasz Garbowski, Kevin Flechsler, Anna Lena Eberle, Gregor Dellemann, Christian Crüger, Wilhelm Bolsinger, Michael Behnke, Pascal Anger, and Dirk Aderhold
T e MultiSEM 505 multi-beam scanning electron microscope (SEM) images large areas at high resolution by increasing the speed at which SEM data can be acquired by almost two orders of magnitude. It uses multiple electron beams in one electron optical column and employs one detection channel for each beam. T e basic principle of operation is the same as with any single-beam SEM, but in the multi-beam SEM the electron source produces an array of 61 electron beams that are focused onto the sample and scanned in parallel 61 small, overlapping subregions of the sample area underneath the beam array. Secondary
electrons emanate from each primary electron spot and are imaged onto a multi-detector with one detection channel for each electron beam. An image of an entire large area is produced by the primary beam array in the time it takes one of the 61 beams to scan its small sub-area. T e maximum scan speed is governed by the minimum number of electrons per pixel required to achieve an acceptable signal-to-noise ratio (SNR), which in turn is limited by the speed and effi ciency of the secondary electron detector. T e electron beam dwell time per pixel is therefore ultimately limited by the bandwidth of the electron detector. Using multiple electron beams in a single column and having a dedicated detector for each beam bypasses the detector bandwidth limit. T e multi-beam electron optical design with its comparably low current per individual single beam also minimizes electron beam damage of the sample. In the past SEMs may not have been considered for applica- tions that require high-throughput imaging. Two important fi elds requiring such high-throughput at nanometer resolution are microelectronics manufacturing and brain research. For example, as structure sizes in semiconductor devices decrease, there is a need for the detection of particles or pattern defects that are only a few nanometers in diameter. A single-beam SEM can sample only a small fraction of wafer area compared to the MultiSEM. For imaging biological samples with electron microscopy, there is an increasing need to image large volumes of biological tissue at high resolution to gain insight into the functioning of human organs. Examples where MultiSEM could have impact include the understanding of neural circuits and the analysis of extended cellular structures.
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