Towards an improved photonic force microscope: a novel technique for biological microscopy

Una delle tecniche più note nello studio topografico di campioni biologici è l’AFM. Ci sono però limitazioni dovute alla presenza del cantilever, il quale pone un limite nella forza minima applicabile su un campione per ottenere un’immagine topografica. Questa forza (ordine dei 10 pN) può essere sufficiente a danneggiare il campione e a deformare i dettagli topografici che si vorrebbero evidenziare. Per superare questo problema si può usare un Photonic Force Microscope, dove il cantilever è sostituito da Optical Tweezers. Questa tecnica permette di effettuare scansioni di campioni biologici applicando forze dell’ordine dei 100 fN. All’interno della trappola ottica viene posizionata una microparticella che agisce da sonda, attraverso la quale possono essere rilevati dettagli topografici del campione. La differenza rispetto al PFM tradizionale si trova proprio nel tipo di sonda utilizzata durante la scansione. Lo standard prevede l’utilizzo di una sonda sferica, di dimensioni dell’ordine dei 100 nm mentre l’ipotesi è che si possano utilizzare delle sonde cilindriche con alla base un dettaglio acuminato che richiama la punta dell’AFM. Questo tipo di sonda consentirebbe di raggiungere una risoluzione maggiore, rispetto al PFM tradizionale, che risente del limite dato dal diametro della sfera. Due differenti setup per la PFM sono stati costruiti e testati durante questo periodo di tesi. Sono state testate diverse microparticelle cilindriche, di dimensioni differenti in termini di aspect ratio con lo scopo di osservare la stabilità di questo tipo di sonda. Nei risultati viene proposto un metodo per controllare la stabilità e l’orientazione della microparticella cilindrica all’interno della trappola ottica. Viene inoltre fatta un’ipotesi su un metodo per stimare la lunghezza della punta che dovrà essere verificata da una misura sistematica. I risultati preliminari riguardanti la scansione di strutture note suggeriscono la validità dell’uso di questo nuovo tipo di sonda

Application of light-sheet mesoscopy to image host-pathogen interactions in intact organs

Human African Trypanosomiasis (HAT) is a disease caused by the extracellular parasite Trypanosoma brucei that affects the central nervous system (CNS) during the chronic stage of the infection, inducing neuroinflammation, coma, and death if left untreated. However, little is known about the structural change happening in the brain as result of the infection. So far, infection-induced neuroinflammation has been observed with conventional methods, such as immunohistochemistry, electron microscopy, and 2-photon microscopy only in small portions of the brain, which may not be representative of the disease. In this paper, we have used a newly-developed light-sheet illuminator to image the level of neuroinflammation in chronically infected mice and compared it to naïve controls. This system was developed for imaging in combination with the Mesolens objective lens, providing fast sub-cellular resolution for tens of mm3-large imaging volumes. The mouse brain specimens were cleared using CUBIC+, followed by antibody staining to locate Glial Fibrillary Acid Protein (GFAP) expressing cells, primarily astrocytes and ependymocytes, used here as a proxy for cell reactivity and gliosis. The large capture volume allowed us to detect GFAP+ cells and spatially resolve the response to T. brucei infection. Based on morphometric analyses and spatial distribution of GFAP+ cells, our data demonstrates a significant increase in cell dendrite branching around the lateral ventricle, as well as dorsal and ventral third ventricles, that are negatively correlated with the branch extension in distal sites from the circumventricular spaces. To our knowledge, this is the first report highlighting the potential of light-sheet mesoscopy to characterise the inflammatory responses of the mouse brain to parasitic infection at the cellular level in intact cleared organs, opening new avenues for the development of new mesoscale imaging techniques for the study of host-pathogen interactions

Challenges and advances in optical 3D mesoscale imaging

Optical mesoscale imaging is a rapidly developing field that allows the visualisation of larger samples than is possible with standard light microscopy, and fills a gap between cell and organism resolution. It spans from advanced fluorescence imaging of micrometric cell clusters to centimetre-size complete organisms. However, with larger volume specimens, new problems arise. Imaging deeper into tissues at high resolution poses challenges ranging from optical distortions to shadowing from opaque structures. This manuscript discusses the latest developments in mesoscale imaging and highlights limitations, namely labelling, clearing, absorption, scattering, and also sample handling. We then focus on approaches that seek to turn mesoscale imaging into a more quantitative technique, analogous to quantitative tomography in medical imaging, highlighting a future role for digital and physical phantoms as well as artificial intelligence

Strategic wine plans: comparative study

El presente trabajo tiene como objetivo comparar doce planes estratégicos para el sector vitivinícola en tres variables: fecha de inicio del plan y horizonte temporal; ámbito de gobernanza: territorial y temático; contenido del plan: objetivos, estrategias planteadas y financiamiento. la metodología es comparativa. en la muestra no probabilística se han seleccionado en forma intencional planes estratégicos provenientes de nueve países y la bibliografía académica sobre ellos. el énfasis está puesto en sintetizar las diferentes opciones o alternativas para la planificación estratégica vitivinícola. El estudio pretende contribuir al estado del arte sobre las políticas para el desarrollo en general y para el sector vitivinícola en particular.This paper aims to compare twelve strategic plans for the wine sector in three variables: start date of the plan and time horizon; governance scope: territorial and thematic; content of the plan: objectives, proposed strategies and financing. The methodology is comparative. the non-probabilistic sample intentionally selected strategic plans from nine countries and the academic literature on them. the emphasis is put on synthesizing the different options or alternatives for strategic wine planning. The study aims to contribute to the state of the art on policies for development in general and for the wine sector in particular.EEA San JuanFil: Hernández, Juan Jesús. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Juan; ArgentinaFil: Novello, Jose Raul. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Juan; Argentina.Fil: Parera, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro Regional Mendoza-San Juan; ArgentinaFil: Perez Peña, Jorge Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; Argentina.Fil: Diaz Bruno, Elda Analia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Santi, Carina Eliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro Regional Mendoza - San Juan; ArgentinaFil: Troilo, Liliana Graciela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Sari, Santiago Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Battistella, Maximiliano. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Juan; Argentin

Light-sheet mesoscopy with the Mesolens provides fast sub-cellular resolution imaging throughout large tissue volumes

Rapid imaging of large biological tissue specimens such as ultra thick sections of mouse brain cannot easily be performed with a standard microscope. Optical mesoscopy offers a solution, but thus far imaging has been too slow to be useful for routine use. We have developed two different illuminators for light-sheet mesoscopy with the Mesolens and we demonstrate their use in high-speed optical mesoscale imaging of large tissue specimens. The first light-sheet approach uses Gaussian optics and is straightforward to implement. It provides excellent lateral resolution and high-speed imaging, but the axial resolution is poor. The second light-sheet is a more complex Airy light-sheet that provides sub-cellular resolution in three dimensions that is comparable in quality to point-scanning confocal mesoscopy, but the light-sheet method of illuminating the specimen reduces the imaging time by a factor of 14. This creates new possibilities for high-content, higher-throughput optical bioimaging at the mesoscale

El Plan Estratégico Vitivinícola 2030 en Argentina = The 2030 Strategic Wine Plan in Argentina

El objetivo del artículo es analizar el proceso de elaboración y las características de la actualización a 2030 del Plan Estratégico Vitivinícola de Argentina (PEVI 2030). Para ello se describen su metodología de redacción, los mecanismos de participación desarrollados, los actores involucrados, los problemas que enfrentó y su contenido. El análisis se realiza en base a bibliografía sobre planificación estratégica en general y para el sector vitivinícola en particular, e identifica la forma en que la prospectiva territorial se incorporó durante el proceso. El artículo concluye que el PEVI 2030 de Argentina es un caso de planificación para el desarrollo de una agroindustria que debe competir en mercados (nacionales e internacionales) y cumplir principios de sostenibilidad. Para la elaboración del Plan se planteó una metodología con mecanismos participativos amplios que encontraron obstáculos por la pandemia, por la reticencia de algunos actores a ser parte del proceso y por la falta de una etapa final de priorización de acciones. Sin embargo, se logró construir una misión y visión prospectiva de una década, con objetivos, estrategias y acciones estratégicas, con un alcance nacional y para todas las cadenas de base vitícola y con una entidad identificada para su implementación.This article has with the objective to analyze the elaboration process and the characteristics of the update to 2030 of the Argentine Vitiviniculture Strategic Plan (PEVI 2030). For this, its writing methodology, the participation mechanisms developed, the actors involved, the problems it faced and its content are described. The analysis is carried out based on bibliography on strategic planning in general and for the wine sector in particular, and identifies the way in which territorial prospective was incorporated during the process. The article concludes that Argentina’s PEVI 2030 is a case of planning for the development of an agro-industry that must compete in national and international markets and comply with sustainability principles. For the elaboration of the Plan, a methodology was proposed with broad participatory mechanisms that encountered obstacles due to the pandemic, due to the reluctance of some actors to be part of the process and due to the lack of a final stage of prioritization of actions. Nonetheless, it was possible to build a mission and prospective vision for the next decade, with objectives, strategies and strategic actions, with national scope and for all wine-based chains and with an entity identified for its implementation.EEA San JuanFil: Hernández, Juan Jesús. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Juan; ArgentinaFil: Santi, Carina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Battistella, Maximiliano. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Juan; Argentina.Fil: Perez Peña, Jorge Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Parera, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Dirección Nacional; Argentin