Next, we tested whether our new reporters can be also used for monitoring rhythms in live preparations.
We tested the method in live preparations of pig left and right ventricular myocardium (thickness 8-18 mm) and phantoms imitating the optical properties of myocardial tissue.
PL cells were present in small numbers in live preparations of SG spillout (Movies S1 and S5), together with metacyclics and other unattached trypanosomes.
In live preparations, we were able to localize the origin of the depolarization wave with a precision of ±1.3mm in the transmural direction and 3 mm in the image plane.
Amongst live preparations, a P + 20% preparation after additional 20 hr in culture looked qualitatively and quantitatively identical to fresh preparation at P + 40%; similarly a P + 40% preparation after additional 15 hr in culture looked like a fresh preparation at P + 55%.
Time-lapse visualization of growing dendrites in live preparations has shown that establishment of a dendritic tree is governed by a combination of local branch dynamics, including elongation, branching and retraction (Dailey and Smith, 1996; Portera-Cailliau et al., 2003; Wu and Cline, 2003; Emoto et al., 2004; Williams and Truman, 2004; Mumm et al., 2006).
Through a prolonged time-course of larval development we vitally stain the mineralized bone and image the living preparation at high resolution by confocal microscopy.
Two-photon microscopy has revolutionized life sciences by enabling long-term imaging of living preparations in highly scattering tissue while minimizing photodamage.
Interestingly, incubations of living preparations of the host-adapted serovar Lai and the culture-adapted serovars Lai and Bataviae resulted in a similar response of the THP-1 cells when compared to the heat-killed response with these bacteria.
Such non-invasive visualization in living preparations allowed us to acquire high-magnification, sequential optical sections (z-stacks) that were used to generate high-resolution anatomical, volumetric images of embryos.
