Abstract
Precise sample orientation is crucial for microscopy but is often performed with macroscopic tools and low accuracy. In vivo imaging of growing and developing samples even requires dynamic adaptation of the sample orientation to continuously achieve optimal imaging. Here, we present a method for freely positioning a sample in 3D by introducing magnetic beads and applying a magnetic field. We demonstrate magnetic orientation of fixed mouse embryos and artemia, and live zebrafish embryos and larvae on an epi-fluorescence microscope and on a light-sheet system for optimal imaging.
Highlights
Precise sample orientation is crucial for microscopy but is often performed with macroscopic tools and low accuracy
We show that after the insertion of magnetic beads, the sample can be freely positioned in 3D during the experiment by applying a magnetic field
In the case of the zebrafish embryo, superparamagnetic beads could be directly injected into the yolk with a microinjection device (Fig. 1c and Supplementary Method 2)
Summary
Precise sample orientation is crucial for microscopy but is often performed with macroscopic tools and low accuracy. To study living organisms under optically optimal conditions, a technique to dynamically adjust sample orientation in the microscope is needed. Sample orientation techniques have been developed for highthroughput applications using microfluidic systems[1] and for analysis of expression patterns in zebrafish larval brains by manually inverting the sample holder[2]. These methods still lack adaptive reorientation of the sample in the microscope during the experiment. In some implementations of light-sheet microscopy (or Selective Plane Illumination Microscopy (SPIM)6) a single degree of adaptive orientation of the sample is offered by a vertical rotational axis (Fig. 1a).
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