Soft X-ray induced radiation damage in thin freeze-dried brain samples studied by FTIR microscopy

In order to push the spatial resolution limits to the nanoscale, synchrotron-based soft X-ray microscopy (XRM) experiments require higher radiation doses to be delivered to materials. Nevertheless, the associated radiation damage impacts on the integrity of delicate biological samples. Herein, the extent of soft X-ray radiation damage in popular thin freeze-dried brain tissue samples mounted onto Si3N4 membranes, as highlighted by Fourier transform infrared microscopy (FTIR), is reported. The freeze-dried tissue samples were found to be affected by general degradation of the vibrational architecture, though these effects were weaker than those observed in paraffin-embedded and hydrated systems reported in the literature. In addition, weak, reversible and specific features of the tissue–Si3N4 interaction could be identified for the first time upon routine soft X-ray exposures, further highlighting the complex interplay between the biological sample, its preparation protocol and X-ray probe

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Soft X-ray induced radiation damage in thin freeze-dried brain samples studied by FTIR microscopy

In order to push the spatial resolution limits to the nanoscale, synchrotron-based soft X-ray microscopy (XRM) experiments require higher radiation doses to be delivered to materials. Nevertheless, the associated radiation damage impacts on the integrity of delicate biological samples. Herein, the extent of soft X-ray radiation damage in popular thin freeze-dried brain tissue samples mounted onto Si3N4 membranes, as highlighted by Fourier transform infrared microscopy (FTIR), is reported. The freeze-dried tissue samples were found to be affected by general degradation of the vibrational architecture, though these effects were weaker than those observed in paraffin-embedded and hydrated systems reported in the literature. In addition, weak, reversible and specific features of the tissue–Si3N4 interaction could be identified for the first time upon routine soft X-ray exposures, further highlighting the complex interplay between the biological sample, its preparation protocol and X-ray probe

Chapter 9: Shock and Offence Online: The Role of Emotion in Participant Absent Research

Purpose - This chapter considers the implications of online participant absent research, that is where the researcher has no interaction with the author, on researcher emotion. Drawing on a study of data extracts ‘mined’ from the Internet without interaction with the author, lessons for practice are identified. It is argued that researchers should reflexively consider the ways in which passive data collection techniques may impact the researcher prior to undertaking research, in order to ensure that researchers are adequately supported. Methodology/approach –The data described in this example were constructed around a single case study where a woman had been asked to leave a sports shop in the UK because she was breastfeeding. Not allowing breastfeeding within a business is illegal in the UK, and resulted in a protest. The study involved analysis of user-generated data from an online news site and Twitter in relation to this case. Findings - Drawing on field notes and conversations with colleagues, I argue that in order to successfully manage my emotional response to the data, it was necessary to distance myself during analysis. Originality/value – Whilst the role of emotion is often considered as part of ethnographic practice in studies utilising face-to-face data collection, it is underexplored in the online domain. This chapter presents, through a detailed example, a reflective account of the emotion work required in one such study. Strategies to reflexively manage emotion for those undertaking participant absent research, which are applicable to both online and off-line situations, are highlighted

We were here, and we still are : negotiations of political space through unsanctioned art

In this chapter, we examine the work of the Sámi artist Anders Sunna and the Egyptian artist Bahia Shebab in order to address strategies of artistic criticism of the relations between states and their citizens. Both artists are protesting against contemporary processes relating to space, state and nation, and they express themselves in ways that are embedded in the aesthetics of unsanctioned street art. This expression constitutes an interesting form of politics, situated somewhere in-between, or alongside, party politics and the practices of civil society. Our aim is to describe and discuss what we see as specifically effective and dynamic themes in the chosen artwork — the use of space as object and methodology, and the production of iconic imageries within fantasies of protest. The stencils and spray paintings of Shehab and Sunna offer us keys to exploring efforts to artistically reveal and dismantle national and neocolonial power.The futures of genders and sexualities. Cultural products, transnational spaces and emerging communitie