Tackling the Mouse‐on‐Mouse Problem in Cochlear Immunofluorescence: A Simple Double‐Blocking Protocol for Immunofluorescent Labeling of Murine Cochlear Sections with Primary Mouse Antibodies

The mouse is the most widely used animal model in hearing research. Immunohistochemistry and immunofluorescent staining of murine cochlear sections have, thus, remained a backbone of inner ear research. Since many primary antibodies are raised in mouse, the problem of "mouse-on-mouse" background arises due to the interaction between the anti-mouse secondary antibody and the native mouse immunoglobulins. Here, we describe the pattern of mouse-on-mouse background fluorescence in sections of the postnatal mouse cochlea. Furthermore, we describe a simple double-blocking immunofluorescence protocol to label mouse cochlear cryosections. The protocol contains a conventional blocking step with serum, and an additional blocking step with a commercially available anti-mouse IgG blocking reagent. This blocking technique virtually eliminates the "mouse-on-mouse" background in murine cochlear sections, while adding only a little time to the staining protocol. We provide detailed instructions and practical tips for tissue harvesting, processing, and immunofluorescence-labeling. Further protocol modifications are described, to shorten the duration of the protocol, based on the primary antibody incubation temperature. Finally, we demonstrate examples of immunofluorescence staining performed using different incubation times and various incubation temperatures with a commercially available mouse monoclonal primary antibody

Ideology in the Era of Xi Jinping

Publisher Copyright: © 2018, The Author(s).After 1978, Maoism as a living mass ideological and social force in the People’s Republic of China largely died away. The Party state’s legitimacy since that time has been based on a new pillar of economic competence and the delivery of tangible economic gains. But China is still a place where, at least within the political elite, there is an identifiable ideology and associated language that links the aims of a political force, the Communist Party of China, with national prosperity, historic rejuvenation, and the delivery of the political goals promised when the Communist Party was founded almost a century ago – modernity in Chinese society. Ideology has not disappeared in this interpretation. It has just become more concealed, more nuanced, and in some spaces more flexible. For Chinese contemporary leaders, ideology is partly a body of practices, beliefs, and language which have been bequeathed to them by previous leaders, and which show that they are part of the same historic movement that runs from 1921 to 1949, and through 1978 until today. This body of practices is aimed at maintaining a sustainable system of one party rule, as well as an assertion of discipline and control in the core tactical spaces of political power. Under Xi, a group of twelve keywords maps out the discursive space that matters to the CPC today. These terms exemplify the ways in which the contemporary CPC is willing to use ideas from diverse sources, either from its own past, or from classical Chinese thinking, as a means of achieving emotional as well as intellectual impact, and to assist in the delivery of the major Party goal of the twenty-first century – the creation of a great nation with the CPC at the heart of its governance. Underlying the keywords and the ideological space they define is the larger notion of the Party, not just attending to material but also spiritual needs – and creating not just a wealthy country, but also a spiritual socialist civilization.publishersversionPeer reviewe

Multi-messenger observations of a binary neutron star merger

On 2017 August 17 a binary neutron star coalescence candidate (later designated GW170817) with merger time 12:41:04 UTC was observed through gravitational waves by the Advanced LIGO and Advanced Virgo detectors. The Fermi Gamma-ray Burst Monitor independently detected a gamma-ray burst (GRB 170817A) with a time delay of ~1.7 s with respect to the merger time. From the gravitational-wave signal, the source was initially localized to a sky region of 31 deg2 at a luminosity distance of 40+8-8 Mpc and with component masses consistent with neutron stars. The component masses were later measured to be in the range 0.86 to 2.26 Mo. An extensive observing campaign was launched across the electromagnetic spectrum leading to the discovery of a bright optical transient (SSS17a, now with the IAU identification of AT 2017gfo) in NGC 4993 (at ~40 Mpc) less than 11 hours after the merger by the One- Meter, Two Hemisphere (1M2H) team using the 1 m Swope Telescope. The optical transient was independently detected by multiple teams within an hour. Subsequent observations targeted the object and its environment. Early ultraviolet observations revealed a blue transient that faded within 48 hours. Optical and infrared observations showed a redward evolution over ~10 days. Following early non-detections, X-ray and radio emission were discovered at the transient’s position ~9 and ~16 days, respectively, after the merger. Both the X-ray and radio emission likely arise from a physical process that is distinct from the one that generates the UV/optical/near-infrared emission. No ultra-high-energy gamma-rays and no neutrino candidates consistent with the source were found in follow-up searches. These observations support the hypothesis that GW170817 was produced by the merger of two neutron stars in NGC4993 followed by a short gamma-ray burst (GRB 170817A) and a kilonova/macronova powered by the radioactive decay of r-process nuclei synthesized in the ejecta

Against the epistemicide. : Itinerant curriculum theory and the reiteration of an epistemology of liberation

Echoing Ettore Scola metaphor “Bruti, Sporchi & Cativi”, this chapter challenges how hegemonic and specific (or so called) counter hegemonic curriculum platforms – so connected with Western Eurocentric Modernity – have been able to colonize the field without any prudency to “fabricate” and impose a classed, raced and gendered philosophy of praxis, as unique, that drives the field to an ideological surrealism and collective suicide. Such collective suicide framed by a theoretical timesharing unleashed by both dominant and specific counter dominant platforms that tenaciously controlled the circuits of cultural production grooms the field as a ghetto, flooded with rudeness, and miserable ambitions, a theoretical caliphate that wipes out any episteme beyond the Western Eurocentric Modern terrain, insolently droving to sewage of society the needs and desires of students, teachers and the community. Drawing from key decolonial thinkers, this chapter examines the way Western eugenic curriculum of modernity created an abyssal thinking in which ‘this side’ of the line is legitimate and ‘the other side’ has been produced as ‘non-existent’ (Sousa Santos B, Another knowledge is possible. Verso, London, 2007). The paper suggests the need to move a post-abyssal curriculum that challenges dominant and counter dominant traditions within ‘this side’ of the line, and respects ‘the other’ side of the line. The paper challenges curriculum studies to assume a non-abyssal position one that respects epistemological diversity. This requires an Itinerant Curriculum Theory (Paraskeva JM, Conflicts in curriculum theory: Challenging hegemonic epistemologies. Palgrave Macmillan, London, 2011), which is a commitment and a ruthless epistemological critique of every existing epistemology

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Multi-messenger Observations of a Binary Neutron Star Merger

On 2017 August 17 a binary neutron star coalescence candidate (later designated GW170817) with merger time 12:41:04 UTC was observed through gravitational waves by the Advanced LIGO and Advanced Virgo detectors. The Fermi Gamma-ray Burst Monitor independently detected a gamma-ray burst (GRB 170817A) with a time delay of ∼ 1.7 {{s}} with respect to the merger time. From the gravitational-wave signal, the source was initially localized to a sky region of 31 deg2 at a luminosity distance of {40}-8+8 Mpc and with component masses consistent with neutron stars. The component masses were later measured to be in the range 0.86 to 2.26 {M}ȯ . An extensive observing campaign was launched across the electromagnetic spectrum leading to the discovery of a bright optical transient (SSS17a, now with the IAU identification of AT 2017gfo) in NGC 4993 (at ∼ 40 {{Mpc}}) less than 11 hours after the merger by the One-Meter, Two Hemisphere (1M2H) team using the 1 m Swope Telescope. The optical transient was independently detected by multiple teams within an hour. Subsequent observations targeted the object and its environment. Early ultraviolet observations revealed a blue transient that faded within 48 hours. Optical and infrared observations showed a redward evolution over ∼10 days. Following early non-detections, X-ray and radio emission were discovered at the transient’s position ∼ 9 and ∼ 16 days, respectively, after the merger. Both the X-ray and radio emission likely arise from a physical process that is distinct from the one that generates the UV/optical/near-infrared emission. No ultra-high-energy gamma-rays and no neutrino candidates consistent with the source were found in follow-up searches. These observations support the hypothesis that GW170817 was produced by the merger of two neutron stars in NGC 4993 followed by a short gamma-ray burst (GRB 170817A) and a kilonova/macronova powered by the radioactive decay of r-process nuclei synthesized in the ejecta.</p