Engineered HaloTag variants for fluorescence lifetime multiplexing

Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce HaloTag variants with either increased or decreased brightness and fluorescence lifetime compared with HaloTag7 when labeled with rhodamines. Combining these HaloTag variants enabled live-cell fluorescence lifetime multiplexing of three cellular targets in one spectral channel using a single fluorophore and the generation of a fluorescence lifetime-based biosensor. Additionally, the brightest HaloTag variant showed up to 40% higher brightness in live-cell imaging applications.

Акустооптический гиперспектральный модуль для гистологического исследования микрообъектов

The problem of the development of an adjustable video-spectrometer unit based on an acousto-optic tunable filter has been considered. This unit is embedded into an optical system of microscopes for hyperspectral analysis of microscopic objects. For this purpose, light–energy conjugation of the filter with microscopes is performed. We have calculated the conjugating optical system, which yields an equality of field of view for a wideband and spectral channel. The experimental model of the developed device has been described. Examples of detection for test objects and histological sections along with the calculation results of the spectral parameters for various skin tissues are given. The developed video-spectrometer unit considerably extends the functionality of microscopes and can be widely used to solve various problems of biomedical diagnostics.

Missing flux in VLBI observations of SiO maser at 7 mm in IRC+10011

VLBI observations of SiO masers recover at most 40-50% of the total flux obtained by single dish observations at any spectral channel. Some previous studies seems to indicate that, at least, part of the lost flux is divided up into many weak components rather than in a large resolved emission area. Taking benefit of the high sensitivity and resolution of the HSA, we investigate the problem of the missing flux in VLBI observations of SiO maser emission at 7 mm in the AGB stars and obtain a high dynamic range map of IRC+10011. We conclude that the missing flux is mostly contained in many very weak maser components.

Differences in liquid cloud droplet effective radius and number concentration estimates between MODIS collections 5.1 and 6 over global oceans

Differences in cloud droplet effective radius and cloud droplet number concentration (CDNC) estimates inferred from the Aqua–MODIS (Moderate Resolution Imaging Spectroradiometer) collections 5.1 (C5.1) and 6 (C6) cloud products (MYD06) are examined for warm clouds over global oceans for the year 2008. Individual pixel level retrievals for both collections are aggregated to 1° × 1° and compared globally and regionally for the three main spectral channel pairs used for MODIS cloud optical property retrievals. Comparisons between both collections are performed for cases in which all three effective radii retrievals are classified by the MODIS cloud product as valid. The contribution to the observed differences of several key MYD06 Collection 6 algorithm updates are also explored, with a focus on changes to the surface reflectance model, assumed solar irradiance, above-cloud emission, cloud-top pressure (CTP), and pixel registration. Global results show a neutral to positive (> 50 cm−3) change for C6-derived CDNC relative to C5.1 for the 1.6 and 2.1 µm channel retrievals, corresponding to a neutral to −2 µm difference in droplet effective radius (re). For 3.7 µm retrievals, CDNC results show a negative change in the tropics, with differences transitioning toward positive values with increasing latitude spanning −25 to +50 cm−3 related to a +2.5 to −1 µm transition in effective radius. Cloud optical thickness (τ) differences were small relative to effective radius and found to not significantly impact CDNC estimates. Regionally, the magnitude and behavior of the annual CDNC cycle are compared for each effective radius retrieval. Results from this study indicate significant inter-collection differences in aggregated values of effective radius due to changes to the precomputed retrieval lookup tables (LUTs) for ocean scenes, changes to retrieved cloud-top pressure, solar irradiance, or above-cloud thermal emission, depending upon spectral channel. The observed differences between collections may have implications for existing MODIS-derived climatologies and validation studies of effective radius and CDNC.