Cell sorting in a Petri dish controlled by computer vision.

Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50-70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66 +/- 12% and 88 +/- 16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95 +/- 2% and 62 +/- 7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS

Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Spontaneous intracellular calcium activity can be observed in a variety of cell types and is proposed to play critical roles in a variety of physiological processes. In particular, appropriate regulation of calcium activity patterns during embryogenesis is necessary for many aspects of vertebrate neural development, including proper neural tube closure, synaptogenesis, and neurotransmitter phenotype specification. While the observation that calcium activity patterns can differ in both frequency and amplitude suggests a compelling mechanism by which these fluxes might transmit encoded signals to downstream effectors and regulate gene expression, existing population-level approaches have lacked the precision necessary to further explore this possibility. Furthermore, these approaches limit studies of the role of cell-cell interactions by precluding the ability to assay the state of neuronal determination in the absence of cell-cell contact. Therefore, we have established an experimental workflow that pairs time-lapse calcium imaging of dissociated neuronal explants with a fluorescence in situ hybridization assay, allowing the unambiguous correlation of calcium activity pattern with molecular phenotype on a single-cell level. We were successfully able to use this approach to distinguish and characterize specific calcium activity patterns associated with differentiating neural cells and neural progenitor cells, respectively; beyond this, however, the experimental framework described in this article could be readily adapted to investigate correlations between any time-series activity profile and expression of a gene or genes of interest

HI Fluctuations at Large Redshifts: II - the Signal Expected for GMRT

For the GMRT, we calculate the expected signal from redshifted HI emission at two frequency bands centered at 610 and 325 MHz. The study focuses on the visibility-visibility cross-correlations, proposed earlier as the optimal statistical estimator for detecting and analyzing this signal. These correlations directly probe the power spectrum of density fluctuations at the redshift where the radiation originated, and thereby provide a method for studying the large scale structures at large redshifts. We present detailed estimates of the correlations expected between the visibilities measured at different baselines and frequencies. Analytic fitting formulas representing the salient features of the expected signal are also provided. These will be useful in planning observations and deciding an optimal strategy for detecting this signal.Comment: 16 pages including 7 figures, published in JAp

Effect of photoperiod and host distribution on the horizontal transmission of Isaria fumosorosea (Hypocreales: Cordycipitaceae) in greenhouse whitefly assessed using a novel model bioassay

A model bioassay was used to evaluate the epizootic potential and determine the horizontal transmission efficiency of Isaria fumosorosea Trinidadian strains against Trialeurodes vaporariorum pharate adults under optimum conditions (25±0.5°C, ~100% RH) at two different photoperiods. Untreated pharate adults were arranged on laminated graph paper at different distributions to simulate varying infestation levels on a leaf surface. Four potential hosts were located 7, 14 and 21 mm away from a central sporulating cadaver simulating high, medium and low infestation levels, respectively. Percent hosts colonized were recorded 7, 12, 14 and 21 days post-treatment during a 16- and 24-h photophase. After 21 days, mean percent hosts colonized at the highest, middle and lowest infestation levels were 93 and 100%, 22 and 58%, 25 and 39% under a 16- and 24-h photophase, respectively. From the results, it was concluded that the longer the photophase, the greater the percentage of hosts colonized, and as host distance increased from the central sporulating cadaver, colonization decreased. The use of this novel model bioassay technique is the first attempt to evaluate the epizootic potential and determine the horizontal transmission efficiency of I. fumosorosea Trinidadian strains under optimal environmental conditions at different photoperiods. This bioassay can be used to assess horizontal transmission efficiency for the selection of fungi being considered for commercial biopesticide development

Decrease Of Repeated Contamination Of Packed Deli-cious Meat Products

The study considers a problem of repeated contamination of delicious products, ready for consumption. The aim of the work is to study the repeated thermal processing of a ready vacuum-packed whole-muscular meat product for inhibiting a surface microbiota.Today it is urgent for the meat industry, because it influences safety and quality, and also limits a storage term of a product.After bringing a meat product to culinary readiness by thermal processing, it has an unessential amount of microbiota. Microorganisms, including pathogenic and conventionally pathogenic ones, fall on a product after its cooking at cutting, preparation to package and at the package stage itself. Microbiological contamination of a ready meat product results in fast spoilage and is a serious problem for producers, because the microbiota growth shortens its storage life. In its turn, it results in a refuse of a consumer to buy this product and great economic losses for producers.The study is directed on a possibility of solving a problem of contamination of a whole-muscular delicious meat product. The solution is in package of a ready product under vacuum and short-term heating at a high temperature.The work is devoted to the complex study of an influence of repeated pasteurization on safety and quality of a product. There was studied an influence of the repeated thermal processing (post-pasteurization) on microbiological, physical-chemical and also organoleptic parameters of a delicious meat product.The special attention is paid to an influence of post-pasteurization regimes on a microbiological condition of studied samples. Studies of a total amount of microbiota and also the presence of sanitary-representative microorganisms were conducted.It has been proven, that the use of post-pasteurization essentially inhibits a number of microorganisms, and also doesn't influence physical-chemical parameters outlook of a product and organoleptic characteristics.Based on studying an influence of post-pasteurization, it has been established, that inhibition of a microbiota essentially influences safety and prolongs the storage term of a product

The complexity of mesoporous silica nanomaterials unravelled by single molecule microscopy

Mesoporous silica nanomaterials are a novel class of materials that offer a highly complex porous network with nanometre-sized channels into which a wide amount of differently sized guests can be incorporated. This makes them an ideal host for various applications for example in catalysis, chromatography and nanomedicine. For these applications, analyzing the host properties and understanding the complicated host–guest interactions is of pivotal importance. In this perspective we review some of our recent work that demonstrates that single molecule microscopy techniques can be utilized to characterize the porous silica host with unprecedented detail. Furthermore, the single molecule studies reveal sample heterogeneities and are a highly efficient tool to gain direct mechanistic insights into the host–guest interactions. Single molecule microscopy thus contributes to a thorough understanding of these nanomaterials enabling the development of novel tailor-made materials and hence optimizing their applicability significantly

Characteristics of a future aeronautical satellite communications system

A possible operational system scenario for providing satellite communications services to the future aviation community was analyzed. The system concept relies on a Ka-band (20/30 GHz) satellite that utilizes multibeam antenna (MBA) technology. The aircraft terminal uses an extremely small aperture antenna as a result of using this higher spectrum at Ka-band. The satellite functions as a relay between the aircraft and the ground stations. The ground stations function as interfaces to the existing terrestrial networks such as the Public Service Telephone Network (PSTN). Various system tradeoffs are first examined to ensure optimized system parameters. High level performance specifications and design approaches are generated for the space, ground, and aeronautical elements in the system. Both technical and economical issues affecting the feasibility of the studied concept are addressed with the 1995 timeframe in mind