ApoTome - Carl Zeiss International

16
The Function Factor: Optical Sections by Means
of Structured Illumination.
ApoTome has continuously proven its leadership position in Fluorescence Imaging
and has received numerous awards from specialist groups. The functional principle
used by ApoTome to generate optical sections is structured illumination, which is so
well-engineered that it provides artifact-free images.
ApoTome: the grid pattern creates clarity
In the ApoTome mode, a grid structure is inserted into the
light field diaphragm plane of the reflected-light beam
path (page 17, Fig. 1A). The grid structure is precisely imaged
into the object plane; it is clearly visible through the
eyepieces and in the live image. In a second step, a highprecision
scanning mechanism moves the grid pattern in
defined steps across the sample plane. In the third step,
images are acquired at each grid position (Figs. 1A-C).
Finally, a single resulting image is calculated from the
three raw images using a fast mathematical algorithm
(Fig. 1D). The result is a precise optical section through
the specimen with no blurring, with elevated contrast,
and perfect for 3D reconstructions.
Precisely in focus: the calibration
A fundamental challenge in generating precise, deblurred
optical sections using structured illumination is to precisely
focus the grid for the selected objective and fluorescence
wavelength. This is very easy with ApoTome.
You start an interactive assistant in AxioVision, and it
An interactive dialog guides you once through the calibration
of the grid focus
guides you through the calibration process step by step.
And in the apochromatic fluorescence beam path of
Axio Imager and Axio Observer, the grid only has to be
shifted minimally or not at all for the different fluorescent
wavelengths.
Interior view of the ApoTome slider: iris diaphragm in position 1, grid and planparallel glass plate in position 2