Apotome.2  - Optical Sectioning Using Structured Illumination
Apotome.2

Apotome.2

Optical sectioning using structured illumination

Create optical sections of your fluorescent samples – free of scattered light. With structured illumination, you know that only the focal plane appears in your image: Apotome.2 recognizes the magnification and moves the appropriate grid into the beampath. The system then calculates your optical section from three images with different grid positions without time lag. It’s a totally reliable way to prevent scattered out-of-focus light, even in your thicker specimens. Operate your system just as easy as always. You get images with high contrast in the best possible resolution – simply brilliant optical sections.

 


The tutorial initializes with an animation of the ZEISS ApoTome in action featured in the upper left-hand side of the window. Beneath the ApoTome model is a cutaway view of the light path showing the aperture grid and the rotating optical glass plate that directs a projection of the grid across the specimen. Presented in the Preview Window is a projection of the grid on the specimen, as would be observed in the software. In order to operate the tutorial, use the ApoTome Position buttons to toggle the ApoTome grid in (Position 2) and out (Position 1) of the light path. When the ApoTome is inserted into the light path, the Grid Projection buttons can be used to manually control the projection position through the use of the Grid Projection Control slider. After the grid is removed using the position control buttons, an aperture diaphragm is slid into position whose size can be varied with the Iris Diaphragm Control slider.

For more details go to the ZEISS Online Campus

Tutorial: Structured Illumination with ZEISS Apotome
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Highlights

Perfect Images – with All Magnifications

Because your applications need different objectives, you need a system that gives you the best resolution for each one. Apotome.2 automatically uses the right grid for your objective, selecting from three grids with different frequencies. With a defined optical section thickness in the region of a Rayleigh unit, you profit from simply brilliant images.

Optimum Results – Free Choice of Light Source and Dyes

From conventional HBO illumination to adjustment free metal-halide lamp HXP 120 C to Colibri.2, the LED illumination source that is gentle on your samples: with Apotome.2 you use exactly the light you need. Apotome.2 also gives you the choice of fluorophores. Whether you work with DAPI, FITC, Rhodamin, Cy5 or with vital dyes such as GFP or mRFP, it’s your decision, not the technology’s. Just change the filter and your system automatically moves the grid to the correct position. From DAPI to Cy5, you get perfect optical sections for multichannel imaging.

 

 

Brilliant Images – Even with Thick Specimens

Your optical section thickness is close to one Rayleigh unit, a value that stands for high axial resolution with a good signal-to-noise ratio. Apotome.2 increases the resolution in Z direction compared to conventional fluorescence microscopy: you obtain brilliant optical sections that allow 3D-rendering , even from thick specimens.

 

 

Applications

Optical sectioning (right) comparison with widefield microscopy (left).
Mouse embryo, tissue section, green: GFP, red: Cy3.
Objective: Plan APOCHROMAT 40 x/1.3 Oil.
Courtesy of N. Büttner, T. Vogel, Centre for Anatomy, University of Göttingen, Germany.

 

Rat hippocampus, triple fluorescence, maximum projection of 3D image stack.
Objective: Plan-APOCHROMAT 63x/1.4.
Courtesy of E. Fuchs & S. Bauch, DPZ Göttingen, Germany.

 

Drosophila embryo, green: HRP, red: glia marker, 100µm Z-stack.
Courtesy of C. Klämbt, Institute for Neurobiology, University of Münster, Germany.

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Downloads

ZEISS Apotome.2

Optical Sections in Fluorescence Imaging

Pages: 16
Filesize: 6,688 kB

Product Information

For the iBooks version: (interactive iBook; 119 MB)

Either download the file to your computer and perform a manual sync with iTunes, or download the file directly onto your iOS device via WiFi and the Safari Browser.

 

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