Tian
Lin
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Optimization of a GCaMP calcium indicator for neural activity imaging
Stable Isotope clues to the formation and evolution of refrozen melt ponds on Arctic Sea ice.
Tian
Lin
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Article
Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Recent efforts in protein engineering have significantly increased the performance of GECIs. The state-of-the art single-wavelength GECI, GCaMP3, has been deployed in a number of model organisms and can reliably detect three or more action potentials in short bursts in several systems in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro assays, we have increased the dynamic range of GCaMP3 by severalfold, creating a family of “GCaMP5” sensors. We tested GCaMP5s in several systems: cultured neurons and astrocytes, mouse retina, and in vivo in Caenorhabditis chemosensory neurons, Drosophila larval neuromuscular junction and adult antennal lobe, zebrafish retina and tectum, and mouse visual cortex. Signal-to-noise ratio was improved by at least 2- to 3-fold. In the visual cortex, two GCaMP5 variants detected twice as many visual stimulus-responsive cells as GCaMP3. By combining in vivo imaging with electrophysiology we show that GCaMP5 fluorescence provides a more reliable measure of neuronal activity than its predecessor GCaMP3. GCaMP5 allows more sensitive detection of neural activity in vivo and may find widespread applications for cellular imaging in general.
Article
Sea ice is one of the determining parameters of the climate system. The presence of melt ponds on the surface of Arctic sea ice plays a critical role in the mass balance of sea ice. A total of nine cores was collected from multiyear ice refrozen melt ponds and adjacent hummocks during the 2015 Arctic Sea State research cruise. The depth profiles of water isotopes, salinity, and ice texture for these sea ice cores were examined to provide information about the development of refrozen melt ponds and water balance generation processes, which are otherwise difficult to acquire. The presence of meteoric water with low oxygen isotope values as relatively thin layers indicates melt pond water stability and little mixing during formation and refreezing. The hydrochemical characteristics of refrozen melt pond and seawater depth profiles indicate little snowmelt enters the upper ocean during melt pond refreezing. Due to the seasonal characters of deuterium excess for Arctic precipitation, water balance calculations utilizing two isotopic tracers (oxygen isotope and deuterium excess) suggest that besides the melt of snow cover, the precipitation input in the melt season may also play a role in the evolution of melt ponds. The dual‐isotope mixing model developed here may become more valuable in a future scenario of increasing Arctic precipitation. The layers of meteoric origin were found at different depths in the refrozen melt pond ice cores. Surface topography information collected at several core sites was examined for possible explanations of different structures of refrozen melt ponds.