Discussion


In summary, we have shown a non-contact sample orientation method that can orient zebrafish embryos and larvae in three-dimensions by applying an external magnetic field after the injection of super-paramagnetic beads into the yolk of the zebrafish embryo.  We optimized the protocol for zebrafish embryos and larvae and adapted the method to a custom-built SPIM and a commercial stereoscope to enable multi-axes multi-view imaging. A major contribution of this method is to enable multi-view imaging irrespective of the microscope used.
There are several applications where this method will be very useful: High-throughput methods Gualda 2015, which do not offer rotational freedom of the sample, will greatly profit from our non-contact sample orientation technique enabling the alignment of all samples in the same orientation potentially increasing the screening sensitivity and specificity. Similarly, laser-cutting experiments need a certain sample orientation for precise cutting of the tissue. Our magnetic manipulator provides full flexibility to orient the sample as desired in such experiments.