A simple approach for multicolor immunofluorescence staining in different Drosophila cell types

Multicolor immunostaining analysis is often a desirable tool in cell biology for most researchers. Nonetheless, this is not an easy task and often not affordable by many laboratories as it might require expensive instrumentation and sophisticated analysis software. Here, we describe a simple protocol for performing sequential immunostainings on two different Drosophila specimens. Our strategy relies on an efficient and reproducible method for removal primary antibodies and/or fluorophore-conjugated secondary antibodies that does not affect antigene integrity. We show that alternation of multiple rounds of antibody incubation and removal on the same slide, followed by registration of the same DAPI-stained image, provides a simple framework for the sequential detection of several antigens in the same cell. Given that the sample fixation procedures used for Drosophila tissues are compatible with most specimen processing protocols, we can envisage that the multicolor immunostaining strategy presented here can be also adapted to different samples including mammalian tissues and/or cells. J. Cell. Physiol. 229: 683-687, 2014. (c) 2013 Wiley Periodicals, Inc

Metrological assessment of TDR performance for moisture evaluation in granular materials

The monitoring of moisture levels and the investigation of the dielectric properties of materials are tasks that can be suitably performed through Time Domain Reflectometry (TDR) techniques. In particular, moisture monitoring of granular agro-foods is a major interest for several industrial applications. Despite the extensive literature regarding the use of TDR technology, the state of the art is lacking in a rigorous metrological characterization of the results achieved, especially for the agro-foods area. To fill this gap, in this paper the Authors carry out a comparative analysis between moisture content measurements obtained through two different TDR instruments on various granular materials. First, the major error contributions affecting the overall performance of the TDR-based method are discussed; in particular, the error contributions that are intrinsic in the method are discriminated from those related to the specific experimental conditions. Successively, systematic errors effects are minimized through a preliminary calibration procedure of the experimental set-up. Finally, results are compared and analyzed through a statistical approach, providing both an assessment of the repeatability of measurements and a rigorous metrological characterization. This ultimately leads to a performance characterization, particularly useful for practical industrial applications

TDR Moisture Estimation for Granular Materials: An Application in Agro-food Industrial Monitoring

Time-domain reflectometry (TDR) is a wellestablished technique for continuous monitoring of the moisture levels and dielectric properties of materials; typically, it is widely used in soil science applications. Despite the high flexibility, elevated accuracy, and the low cost of TDR instrumentation, the state of the art is actually rather lacking in specific applications for the quality monitoring of agro-industrial processes and agro-food products. Therefore, the possibility of real-time control of moisture level plays a crucial role in terms of optimization, quality preservation, and energy saving in several industrial applications. In this regard, the development of suitable models relating the\ud dielectric properties of granular materials to their moisture content is a still an open issue. To fill this gap, in this paper, the authors study the feasibility of adopting the TDR technique for such a purpose. The proposed goal is successfully reached through a comparative analysis among different calibration procedures and through experimental measurements on various granular materials. Finally, the adopted methodology is assessed through a rigorous metrological characterization, thus allowing the evaluation of the associated measurement uncertainty and repeatability

TDR Moisture Measurements in Granular Materials: from the Siliceous Sand test-case to the Applications for Agro-food Industrial Monitoring

Different approaches are commonly available for the continuous monitoring of moisture and dielectric properties of various materials. In particular Time Domain Reflectometry (TDR) is the standard method for measuring soil water content. Nevertheless, the state of the art is rather lacking in assessing specific calibration procedures allowing the accurate moisture monitoring of granular materials. To fill this gap, the adoption of TDR for such purposes is investigated. Starting from measurements on siliceous sand, an in-depth analysis is extended to various feedstuff materials (such as corn, corn flour and bran). Furthermore, the adopted methodology is fully assessed from a metrological point of view, thus confirming its suitability for the considered applications

Metrological assessment of TDR performance for moisture evaluation in granular materials

The monitoring of moisture levels and the investigation of the dielectric properties of materials are tasks that can be suitably performed through Time Domain Reflectometry (TDR) techniques. In particular, moisture monitoring of granular agro-foods is a major interest for several industrial applications. Despite the extensive literature regarding the use of TDR technology, the state of the art is lacking in a rigorous metrological characterization of the results achieved, especially for the agro-foods area. To fill this gap, in this paper the Authors carry out a comparative analysis between moisture content measurements obtained through two different TDR instruments on various granular materials. First, the major error contributions affecting the overall performance of the TDR-based method are discussed; in particular, the error contributions that are intrinsic in the method are discriminated from those related to the specific experimental conditions. Successively, systematic errors effects are minimized through a preliminary calibration procedure of the experimental set-up. Finally, results are compared and analyzed through a statistical approach, providing both an assessment of the repeatability of measurements and a rigorous metrological characterization. This ultimately leads to a performance characterization, particularly useful for practical industrial applications. © 2008 Elsevier Ltd. All rights reserved

A Non-Invasive Approach for Moisture Measurements through Patch Antennas

Different approaches and techniques are commonly available for the continuous monitoring of moisture levels and dielectric properties of materials; typically, related applications, are largely used in soil science area. Nevertheless, the state of the art is actually rather lacking in specific sensing methods that combine the low cost and the non-invasive approach of the involved instrumentation. To fill this gap, in this paper the authors study the feasibility of the adoption of the time-domain reflectometry analysis for such purposes, using a patch antenna as sensing element. This way, the extrapolation of the scattering parameters is suitably correlated to the water content levels of the materials under test. Furthermore, the adopted methodology is assessed through full-wave simulations, thus corroborating the experimental measurements

Phenotypic characterization of diamond (dind), a Drosophila gene required for multiple aspects of cell division

Many genes are required for the assembly of the mitotic apparatus and for proper chromosome behavior during mitosis and meiosis. A fruitful approach to elucidate the mechanisms underlying cell division is the accurate phenotypic characterization of mutations in these genes. Here, we report the identification and characterization of diamond (dind), an essential Drosophila gene required both for mitosis of larval brain cells and for male meiosis. Larvae homozygous for any of the five EMS-induced mutations die in the third-instar stage and exhibit multiple mitotic defects. Mutant brain cells exhibit poorly condensed chromosomes and frequent chromosome breaks and rearrangements; they also show centriole fragmentation, disorganized mitotic spindles, defective chromosome segregation, endoreduplicated metaphases, and hyperploid and polyploid cells. Comparable phenotypes occur in mutant spermatogonia and spermatocytes. The dind gene encodes a non-conserved protein with no known functional motifs. Although the Dind protein exhibits a rather diffuse localization in both interphase and mitotic cells, fractionation experiments indicate that some Dind is tightly associated with the chromatin. Collectively, these results suggest that loss of Dind affects chromatin organization leading to defects in chromosome condensation and integrity, which in turn affect centriole stability and spindle assembly. However, our results do not exclude the possibility that Dind directly affects some behaviors of the spindle and centrosomes