Graham D. Wright, Kerry A. Thompson, Yara Reis, Johanna Bischof, Philip Edward Hockberger, Michelle S. Itano, Lisa Yen, Stephen Taiye Adelodun, Nikki Bialy, Claire M. Brown, Linda Chaabane, Teng-Leong Chew, Andrew I. Chitty, Fabrice P. Cordelières, Mariana De Niz, Jan Ellenberg, Lize Engelbrecht, Eunice Fabian-Morales, Elnaz Fazeli, Julia Fernandez-Rodriguez, Elisa Ferrando-May, Georgina Fletcher, Graham John Galloway, Adan Guerrero, Jander Matos Guimarães, Caron A. Jacobs, Sachintha Jayasinghe, Eleanor Kable, Gregory T Kitten, Shinya Komoto, Xiaoxiao Ma, Jéssica Araújo Marques, Bryan A. Millis, Kildare Miranda, Peter JohnO'Toole, Sunday Yinka Olatunji, Federica Paina, Cora Noemi Pollak, Clara Prats, Joanna W. Pylvänäinen, Mai Atef Rahmoon, Michael A. Reiche, James Douglas Riches, Andres Hugo Rossi, Jean Salamero, Caroline Thiriet, Stefan Terjung, Aldenora dos Santos Vasconcelos, Antje Keppler
In the dynamic landscape of scientific research, imaging core facilities are vital hubs propelling collaboration and innovation at the technology development and dissemination frontier. Here, we present a collaborative effort led by Global BioImaging (GBI), introducing international recommendations geared towards elevating the careers of Imaging Scientists in core facilities. Despite the critical role of Imaging Scientists in modern research ecosystems, challenges persist in recognising their value, aligning performance metrics and providing avenues for career progression and job security. The challenges encompass a mismatch between classic academic career paths and service-oriented roles, resulting in a lack of understanding regarding the value and impact of Imaging Scientists and core facilities and how to evaluate them properly. They further include challenges around sustainability, dedicated training opportunities and the recruitment and retention of talent. Structured across these interrelated sections, the recommendations within this publication aim to propose globally applicable solutions to navigate these challenges. These recommendations apply equally to colleagues working in other core facilities and research institutions through which access to technologies is facilitated and supported. This publication emphasises the pivotal role of Imaging Scientists in advancing research programs and presents a blueprint for fostering their career progression within institutions all around the world.
{"title":"Recognising the importance and impact of Imaging Scientists: Global guidelines for establishing career paths within core facilities","authors":"Graham D. Wright, Kerry A. Thompson, Yara Reis, Johanna Bischof, Philip Edward Hockberger, Michelle S. Itano, Lisa Yen, Stephen Taiye Adelodun, Nikki Bialy, Claire M. Brown, Linda Chaabane, Teng-Leong Chew, Andrew I. Chitty, Fabrice P. Cordelières, Mariana De Niz, Jan Ellenberg, Lize Engelbrecht, Eunice Fabian-Morales, Elnaz Fazeli, Julia Fernandez-Rodriguez, Elisa Ferrando-May, Georgina Fletcher, Graham John Galloway, Adan Guerrero, Jander Matos Guimarães, Caron A. Jacobs, Sachintha Jayasinghe, Eleanor Kable, Gregory T Kitten, Shinya Komoto, Xiaoxiao Ma, Jéssica Araújo Marques, Bryan A. Millis, Kildare Miranda, Peter JohnO'Toole, Sunday Yinka Olatunji, Federica Paina, Cora Noemi Pollak, Clara Prats, Joanna W. Pylvänäinen, Mai Atef Rahmoon, Michael A. Reiche, James Douglas Riches, Andres Hugo Rossi, Jean Salamero, Caroline Thiriet, Stefan Terjung, Aldenora dos Santos Vasconcelos, Antje Keppler","doi":"10.1111/jmi.13307","DOIUrl":"10.1111/jmi.13307","url":null,"abstract":"<p>In the dynamic landscape of scientific research, imaging core facilities are vital hubs propelling collaboration and innovation at the technology development and dissemination frontier. Here, we present a collaborative effort led by Global BioImaging (GBI), introducing international recommendations geared towards elevating the careers of Imaging Scientists in core facilities. Despite the critical role of Imaging Scientists in modern research ecosystems, challenges persist in recognising their value, aligning performance metrics and providing avenues for career progression and job security. The challenges encompass a mismatch between classic academic career paths and service-oriented roles, resulting in a lack of understanding regarding the value and impact of Imaging Scientists and core facilities and how to evaluate them properly. They further include challenges around sustainability, dedicated training opportunities and the recruitment and retention of talent. Structured across these interrelated sections, the recommendations within this publication aim to propose globally applicable solutions to navigate these challenges. These recommendations apply equally to colleagues working in other core facilities and research institutions through which access to technologies is facilitated and supported. This publication emphasises the pivotal role of Imaging Scientists in advancing research programs and presents a blueprint for fostering their career progression within institutions all around the world.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.13307","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140839573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alastair J. McGinness, Susan A. Brooks, Richard Strasser, Jennifer Schoberer, Verena Kriechbaumer
Plant cells are a capable system for producing economically and therapeutically important proteins for a variety of applications, and are considered a safer production system than some existing hosts such as bacteria or yeasts. However, plants do not perform protein modifications in the same manner as mammalian cells do. This can impact on protein functionality for plant‐produced human therapeutics. This obstacle can be overcome by creating a plant‐based system capable of ‘humanising’ proteins of interest resulting in a glycosylation profile of synthetic plant‐produced proteins as it would occur in mammalian systems.For this, the human glycosylation enzymes (HuGEs) involved in N‐linked glycosylation N‐acetylglucosaminyltransferase IV and V (GNTIV and GNTV), β‐1,4‐galactosyltransferase (B4GALT1), and α‐2,6‐sialyltransferase (ST6GAL) were expressed in plant cells. For these enzymes to carry out the stepwise glycosylation functions, they need to localise to late Golgi body cisternae. This was achieved by a protein targeting strategy of replacing the mammalian Golgi targeting domains (Cytoplasmic‐Transmembrane‐Stem (CTS) regions) with plant‐specific ones. Using high‐resolution and dynamic confocal microscopy, we show that GNTIV and GNTV were successfully targeted to the medial‐Golgi cisternae while ST6GAL and B4GALT1 were targeted to trans‐Golgi cisternae.Plant cells are a promising system to produce human therapeutics for example proteins used in enzyme replacement therapies. Plants can provide safer and cheaper alternatives to existing expression systems such as mammalian cell culture, bacteria or yeast. An important factor for the functionality of therapeutic proteins though are protein modifications specific to human cells. However, plants do not perform protein modifications in the same manner as human cells do. Therefore, plant cells need to be genetically modified to mimic human protein modifications patterns. The modification of importance here, is called N‐linked glycosylation and adds specific sugar molecules onto the proteins.Here we show the expression of four human glycosylation enzymes, which are required for N‐linked glycosylation, in plant cells.In addition, as these protein modifications are carried out in cells resembling a factory production line, it is important that the human glycosylation enzymes be placed in the correct cellular compartments and in the correct order. This is carried out in Golgi bodies. Golgi bodies are composed of several defined stacks termed cis‐, medial and trans‐Golgi body stacks. For correct protein function, two of these human glycosylation enzymes need to be placed in the medial‐Golgi attacks and the other two in the trans‐Golgi stacks. Using high‐resolution laser microscopy in live plant cells, we show here that the human glycosylation enzymes are sent within the cells to the correct Golgi body s
{"title":"Suborganellar resolution imaging for the localisation of human glycosylation enzymes in tobacco Golgi bodies","authors":"Alastair J. McGinness, Susan A. Brooks, Richard Strasser, Jennifer Schoberer, Verena Kriechbaumer","doi":"10.1111/jmi.13311","DOIUrl":"https://doi.org/10.1111/jmi.13311","url":null,"abstract":"Plant cells are a capable system for producing economically and therapeutically important proteins for a variety of applications, and are considered a safer production system than some existing hosts such as bacteria or yeasts. However, plants do not perform protein modifications in the same manner as mammalian cells do. This can impact on protein functionality for plant‐produced human therapeutics. This obstacle can be overcome by creating a plant‐based system capable of ‘humanising’ proteins of interest resulting in a glycosylation profile of synthetic plant‐produced proteins as it would occur in mammalian systems.For this, the human glycosylation enzymes (HuGEs) involved in N‐linked glycosylation N‐acetylglucosaminyltransferase IV and V (GNTIV and GNTV), β‐1,4‐galactosyltransferase (B4GALT1), and α‐2,6‐sialyltransferase (ST6GAL) were expressed in plant cells. For these enzymes to carry out the stepwise glycosylation functions, they need to localise to late Golgi body cisternae. This was achieved by a protein targeting strategy of replacing the mammalian Golgi targeting domains (Cytoplasmic‐Transmembrane‐Stem (CTS) regions) with plant‐specific ones. Using high‐resolution and dynamic confocal microscopy, we show that GNTIV and GNTV were successfully targeted to the medial‐Golgi cisternae while ST6GAL and B4GALT1 were targeted to <jats:italic>trans‐</jats:italic>Golgi cisternae.Plant cells are a promising system to produce human therapeutics for example proteins used in enzyme replacement therapies. Plants can provide safer and cheaper alternatives to existing expression systems such as mammalian cell culture, bacteria or yeast. An important factor for the functionality of therapeutic proteins though are protein modifications specific to human cells. However, plants do not perform protein modifications in the same manner as human cells do. Therefore, plant cells need to be genetically modified to mimic human protein modifications patterns. The modification of importance here, is called N‐linked glycosylation and adds specific sugar molecules onto the proteins.Here we show the expression of four human glycosylation enzymes, which are required for N‐linked glycosylation, in plant cells.In addition, as these protein modifications are carried out in cells resembling a factory production line, it is important that the human glycosylation enzymes be placed in the correct cellular compartments and in the correct order. This is carried out in Golgi bodies. Golgi bodies are composed of several defined stacks termed <jats:italic>cis</jats:italic>‐, medial and <jats:italic>trans</jats:italic>‐Golgi body stacks. For correct protein function, two of these human glycosylation enzymes need to be placed in the medial‐Golgi attacks and the other two in the <jats:italic>trans</jats:italic>‐Golgi stacks. Using high‐resolution laser microscopy in live plant cells, we show here that the human glycosylation enzymes are sent within the cells to the correct Golgi body s","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2024-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140839393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nandini Aggarwal, Richard Marsh, Stefania Marcotti, Tanya J Shaw, Brian Stramer, Susan Cox, Siân Culley
SummaryMany biological structures take the form of fibres and filaments, and quantitative analysis of fibre organisation is important for understanding their functions in both normal physiological conditions and disease. In order to visualise these structures, fibres can be fluorescently labelled and imaged, with specialised image analysis methods available for quantifying the degree and strength of fibre alignment. Here we show that fluorescently labelled fibres can display polarised emission, with the strength of this effect varying depending on structure and fluorophore identity. This can bias automated analysis of fibre alignment and mask the true underlying structural organisation. We present a method for quantifying and correcting these polarisation effects without requiring polarisation‐resolved microscopy and demonstrate its efficacy when applied to images of fluorescently labelled collagen gels, allowing for more reliable characterisation of fibre microarchitecture.
{"title":"Characterisation and correction of polarisation effects in fluorescently labelled fibres","authors":"Nandini Aggarwal, Richard Marsh, Stefania Marcotti, Tanya J Shaw, Brian Stramer, Susan Cox, Siân Culley","doi":"10.1111/jmi.13308","DOIUrl":"https://doi.org/10.1111/jmi.13308","url":null,"abstract":"SummaryMany biological structures take the form of fibres and filaments, and quantitative analysis of fibre organisation is important for understanding their functions in both normal physiological conditions and disease. In order to visualise these structures, fibres can be fluorescently labelled and imaged, with specialised image analysis methods available for quantifying the degree and strength of fibre alignment. Here we show that fluorescently labelled fibres can display polarised emission, with the strength of this effect varying depending on structure and fluorophore identity. This can bias automated analysis of fibre alignment and mask the true underlying structural organisation. We present a method for quantifying and correcting these polarisation effects without requiring polarisation‐resolved microscopy and demonstrate its efficacy when applied to images of fluorescently labelled collagen gels, allowing for more reliable characterisation of fibre microarchitecture.","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140839398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sebastian Munck, Christof De Bo, Christopher Cawthorne, Julien Colombelli
Developing devices and instrumentation in a bioimaging core facility is an important part of the innovation mandate inherent in the core facility model but is a complex area due to the required skills and investments, and the impossibility of a universally applicable model. Here, we seek to define technological innovation in microscopy and situate it within the wider core facility innovation portfolio, highlighting how strategic development can accelerate access to innovative imaging modalities and increase service range, and thus maintain the cutting edge needed for sustainability. We consider technology development from the perspective of core facility staff and their stakeholders as well as their research environment and aim to present a practical guide to the ‘Why, When, and How’ of developing and integrating innovative technology in the core facility portfolio.
Core facilities need to innovate to stay up to date. However, how to carry out the innovation is not very obvious. One area of innovation in imaging core facilities is the building of optical setups. However, the creation of optical setups requires specific skill sets, time, and investments. Consequently, the topic of whether a core facility should develop optical devices is discussed as controversial. Here, we provide resources that should help get into this topic, and we discuss different options when and how it makes sense to build optical devices in core facilities. We discuss various aspects, including consequences for staff and the relation of the core to the institute, and also broaden the scope toward other areas of innovation.
{"title":"Innovating in a bioimaging core through instrument development","authors":"Sebastian Munck, Christof De Bo, Christopher Cawthorne, Julien Colombelli","doi":"10.1111/jmi.13312","DOIUrl":"10.1111/jmi.13312","url":null,"abstract":"<p>Developing devices and instrumentation in a bioimaging core facility is an important part of the innovation mandate inherent in the core facility model but is a complex area due to the required skills and investments, and the impossibility of a universally applicable model. Here, we seek to define technological innovation in microscopy and situate it within the wider core facility innovation portfolio, highlighting how strategic development can accelerate access to innovative imaging modalities and increase service range, and thus maintain the cutting edge needed for sustainability. We consider technology development from the perspective of core facility staff and their stakeholders as well as their research environment and aim to present a practical guide to the ‘Why, When, and How’ of developing and integrating innovative technology in the core facility portfolio.</p><p>Core facilities need to innovate to stay up to date. However, how to carry out the innovation is not very obvious. One area of innovation in imaging core facilities is the building of optical setups. However, the creation of optical setups requires specific skill sets, time, and investments. Consequently, the topic of whether a core facility should develop optical devices is discussed as controversial. Here, we provide resources that should help get into this topic, and we discuss different options when and how it makes sense to build optical devices in core facilities. We discuss various aspects, including consequences for staff and the relation of the core to the institute, and also broaden the scope toward other areas of innovation.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.13312","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140839396","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}