Two-Photon Imaging of Calcium in Virally Transfected Striate Cortical Neurons of Behaving Monkey

Two-photon scanning microscopy has advanced our understanding of neural signaling in non-mammalian species and mammals. Various developments are needed to perform two-photon scanning microscopy over prolonged periods in non-human primates performing a behavioral task. In striate cortex in two macaque monkeys, cortical neurons were transfected with a genetically encoded fluorescent calcium sensor, memTNXL, using AAV1 as a viral vector. By constructing an extremely rigid and stable apparatus holding both the two-photon scanning microscope and the monkey's head, single neurons were imaged at high magnification for prolonged periods with minimal motion artifacts for up to ten months. Structural images of single neurons were obtained at high magnification. Changes in calcium during visual stimulation were measured as the monkeys performed a fixation task. Overall, functional responses and orientation tuning curves were obtained in 18.8% of the 234 labeled and imaged neurons. This demonstrated that the two-photon scanning microscopy can be successfully obtained in behaving primates

Habitat conservation in Italy: the state of the art in the light of the first European Red List of Terrestrial and Freshwater Habitats

The importance of taking into account ecosystems, plant communities and habitats for the development of biodiversity conservation strategies is increasingly acknowledged. Recently, the first ever European Red List of Habitats was produced, which provided an evaluation of the extinction risk of EUNIS-based natural and semi-natural habitats in Europe. As assessment unit, it used the habitat intended as a plant community, thus representing a landmark for the role of vegetation science in nature conservation. In the present paper, the results of the European Red List of Habitats are analyzed at the national scale with specific reference to the terrestrial and freshwater habitat types occurring in Italy. More than three-quarters of the assessed European habitat types were recognized for the Italian territory. The distribution of the threat categories reflects approximately the situation at the EU28 level. About 35% of the assessed habitat types are referred to a threat category; no critically endangered habitat is present in Italy. The most frequently used criteria are those related to a reduction in quantity. Some critical issues arising from the analyses are discussed. In particular, the presence of knowledge gaps is pointed out, with remarkable reference to the poor availability of spatial and quantitative data, severely affecting the application of the criteria adopted for the assessment. Descriptions of habitat types from Italy are reported, some of which are representative, emblematic or even exclusive to the Italian territory. The outcomes of the analysis represent the starting point for the future development of a national-scale Red List of Habitats. Results also emphasized how habitat types with a too broad definition pose a limit to a proper evaluation of the regional biogeographic variability, often very high in Italy, with local floristic and phytocoenotic peculiarities which do not find room in the adopted European typology. This is the reason why the development of national subtypes stands as a necessary step for the development of a realistic and effective assessment at the national scale

Positional stability of holographic optical traps

The potential of digital holography for complex manipulation of micron-sized particles with optical tweezers has been clearly demonstrated. By contrast, its use in quantitative experiments has been rather limited, partly due to fluctuations introduced by the spatial light modulator (SLM) that displays the kinoforms. This is an important issue when high temporal or spatial stability is a concern. We have investigated the performance of both an analog-addressed and a digitally-addressed SLM, measuring the phase fluctuations of the modulated beam and evaluating the resulting positional stability of a holographic trap. We show that, despite imparting a more unstable modulation to the wavefront, our digitally-addressed SLM generates optical traps in the sample plane stable enough for most applications. We further show that traps produced by the analog-addressed SLM exhibit a superior pointing stability, better than 1 nm, which is comparable to that of non-holographic tweezers. These results suggest a means to implement precision force measurement experiments with holographic optical tweezers (HOTs)