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Biology subjects

Millot, A.

Publications and source records attributed to Millot, A..

4 recordsLinked to original sources

Open Blink: Low-cost TIRF microscopy for super-resolutionimaging via μManager

Super-resolution localization microscopy (SMLM) has become a central tool for nanoscale biological research for its high spatial resolution and compatibility with wide-field microscopy. Achieving quantitative SMLM, however, requires homogeneous high-power illumination, nanometric axial stability, and precise multi-channel detection, features typically restricted to high-end commercial instruments or custom solutions in specialized laboratories. The cost of such microscopes and their technical complexity still limit the accessibility of these advanced imaging techniques. Several home-made single molecule microscopes and their submodules have been demonstrated as opensource, highly-customizable, and cost-effective alternatives for their commercial counterparts. Yet, implementation of such systems often requires expert knowledge in optics, electronics, and control system engineering. We introduce Open Blink, a compact open-source TIRF microscope integrating powerful homogeneous quad-line laser illumination, dual-channel detection, and active focus-lock stabilization for quantitative multi-color super-resolution imaging. Open Blink achieves a localization precision below 10 nm in dSTORM, supports a tunable, large field of view from 105 x 105 {micro}m2 up to 257 x 257 {micro}m2, and maintains axial stability over hours, enabling high-throughput super-resolution acquisition. Built with predominantly off-the-shelf components, and full integration into the open-source software {micro}Manager where metadata registration ensures reproducibility, Open Blink offers a low threshold for adoption by easing implementation, use and maintenance. At a substantially reduced cost of approximately 70 000 Euros, among which the high-power laser combiner alone is less than 20 000 euros, Open Blink greatly improves accessibility for laboratories who wish to implement scalable high performance super-resolution microscopy based on single molecules.

biophysics↗

Unidirectional response to bidirectional selection on body size. II Quantitative genetics.

Anticipating the genetic and phenotypic changes induced by natural or artificial selection requires reliable estimates of trait evolvabilities (genetic variances and covariances). However, whether or not multivariate quantitative genetics models are able to predict precisely the evolution of traits of interest, especially fitness-related, life-history traits, remains an open empirical question. Here, we assessed to what extent the response to bivariate artificial selection on both body size and maturity in the medaka Oryzias latipes, a model fish species, fits the theoretical predictions. Three lines (Large, Small, and Control lines) were differentially selected for body length at 75 days of age, conditional on maturity. As maturity and body size were phenotypically correlated, this selection procedure generated a bi-dimensional selection pattern on two life history traits. After removal of non-heritable trends and noise with a random effect (animal) model, the observed selection response did not match the expected bidirectional response. For body size, Large and Control lines responded along selection gradients (larger body size and stasis, respectively), but, surprisingly, the Small did not evolve a smaller body length, and remained identical to the Control line throughout the experiment. The magnitude of the empirical response was smaller than the theoretical prediction in both selected directions. For maturity, the response was opposite to the expectation (the Large line evolved late maturity compared to the Control line, while the Small line evolved early maturity, while the opposite pattern was predicted due to the strong positive genetic correlation between both traits). The mismatch between predicted and observed response was substantial and could not be explained by usual sources of uncertainties (including sampling effects, genetic drift, and error in G matrix estimates).

evolutionary biology↗

Density-dependent selection mediates harvest-induced evolution

Rapid life-history changes caused by size-selective harvesting are often interpreted as a response to direct harvest selection against a large body size. However, similar trait changes may result from a harvest-induced relaxation of natural selection for a large body size via density-dependent selection. Here, we show evidence of such density-dependent selection favouring large-bodied individuals at high population densities, in replicated pond populations of medaka fish. Harvesting, in contrast, selected medaka directly against large-bodied medaka and, in parallel, decreased medaka population densities. Five years of harvesting were enough for harvested and unharvested medaka populations to inherit the classically-predicted trait differences, whereby harvested medaka grew slower and matured earlier than unharvested medaka. We demonstrate that this life-history divergence was not driven by direct harvest selection for a smaller body size in harvested populations, but by density-dependent natural selection for a larger body size in unharvested populations.

evolutionary biology↗

Anthropogenic selection along directions of most evolutionary resistance

Anthropogenic perturbations such as harvesting often select against a large body size and are predicted to induce rapid evolution towards smaller body sizes and earlier maturation. However, body-size evolvability and, hence, adaptability to anthropogenic perturbations remain seldom evaluated in wild populations. Here, we use a laboratory experiment over 6 generations to measure the ability of wild-caught medaka fish (Oryzias latipes) to evolve in response to bidirectional size-dependent selection mimicking opposite harvest regimes. Specifically, we imposed selection against a small body size (Large line), against a large body size (Small line) or random selection (Control line), and measured correlated responses across multiple phenotypic, life-history and endocrine traits. As expected, the Large line evolved faster somatic growth and delayed maturation, but also evolved smaller body sizes at hatch, with no change in average levels of pituitary gene expressions of luteinizing, folliclestimulating or growth (GH) hormones. In contrast, the Small medaka line was unable to evolve smaller body sizes or earlier maturation, but evolved smaller body sizes at hatch and showed marginally-significant signs of increased reproductive investment, including larger egg sizes and elevated pituitary GH production. Natural selection on medaka body size was too weak to significantly hinder the effect of artificial selection, indicating that the asymmetric body-size response to size-dependent selection reflected an asymmetry in body-size evolvability. Our results show that trait evolvability may be contingent upon the direction of selection, and that a detailed knowledge of trait evolutionary potential is needed to forecast population response to anthropogenic change.

evolutionary biology↗