Acquisition Parameter - Focus Devices And Strategy - Zeiss LSM 880 Operating Manual

LSM 880
5.2.11 Acquisition Parameter – Focus
Devices and Strategy
Focus Devices and Strategy aims to maintain
focus stability in multi-dimensional acquisition
experiments.
This tool is typically employed to compensate for Z-
drift in time series experiments as well as for the
tilt of samples when recording at multiple positions
using a scanning stage.
Focus Devices and Strategy features four
different autofocus
combination with the acquisition tools Time
Series, Tile Scan and Positions.
These modes employ either the optional module Definite Focus or the built-in detection capabilities of
the LSM.
The set of available autofocus modes therefore depends upon a system's hardware configuration. The
LSM-based autofocus modes Fluorescence, Reflection and Autofocus Map cannot be used if an
acquisition strategy includes tracks with camera-based image recording.
The Focus Devices and Strategy tool (Fig. 169) provides controls for maintaining focus stability with the
Tile Scan and Positions tools.
Autofocus Mode specifies the method to be used throughout the course of a
multi-dimensional experiment.
The menu for mode selection features a maximum of four active modes
(or Off mode):
All LSM-based autofocus modes rely on the optical sectioning capabilities of a LSM for precisely locating
signal intensity maxima in the Z-direction.
The Z-position of an intensity maximum found within the search range is used as a reference for the
positioning of the imaging plane in the Z-direction.
In many cases, reference and imaging plane are not identical but differ by a fixed distance referred to as
offset (see Reflection and Autofocus Map modes below).
Whenever a focusing action is executed, the Z-position of the imaging plane is updated.
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
modes to be used
Fluorescence
−
Reflection
−
Autofocus Map
−
Definite Focus
−
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Fig. 169 Focus Devices and Strategy tool
in
ZEISS
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