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More than 20 years after their discovery, fluorescent proteins (FPs) continue to be the subject of massive engineering efforts yielding continued improvements. Among these efforts are many aspects that should be of great interest to quantitative imaging users. With new variants frequently introduced into the research community, "tried and true" FPs that have been relied on for many years may now be due for upgrades to more modern variants. However, the dizzying array of FPs now available can make the initial act of narrowing down the potential choices an intimidating prospect. This chapter describes the FP properties that most strongly impact their performance in quantitative imaging experiments, along with their physical origins as they are currently understood. A workflow for evaluating a given FP in the researcher's chosen experimental system (e.g., a specific cell line) is described.
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http://dx.doi.org/10.1016/B978-0-12-420138-5.00006-9 | DOI Listing |
Proc Natl Acad Sci U S A
September 2025
Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.
Multivalent binding and the resulting dynamical clustering of receptors and ligands are known to be key features in biological interactions. For optimizing biomaterials capable of similar dynamical features, it is essential to understand the first step of these interactions, namely the multivalent molecular recognition between ligands and cell receptors. Here, we present the reciprocal cooperation between dynamic ligands in supramolecular polymers and dynamic receptors in model cell membranes, determining molecular recognition and multivalent binding via receptor clustering.
View Article and Find Full Text PDFJ Vis Exp
August 2025
Institut de recherches cliniques de Montréal (IRCM); Programmes de biologie moléculaire, Université de Montréal; Département de Médecine, Université de Montréal;
Embryonic tissue growth and patterning are largely controlled by signals exchanged locally between cell populations within the tissues themselves. Cytonemes are a type of signaling filopodia first identified in Drosophila that connect and mediate exchange between signal-producing and signal-receiving cells. In the developing Drosophila wing imaginal disc, cytonemes are involved in signal exchange between distinct populations of cells within the disc proper (DP) epithelium, which will form the adult wing, as well as between DP cells and cells in adjacent disc-associated tissues.
View Article and Find Full Text PDFJ Bacteriol
September 2025
Department of Microbiology and Immunology, Carver College of Medicine, University of Iowa, Iowa City, Iowa, USA.
Essential genes are interesting in their own right and as potential antibiotic targets. To date, only one report has identified essential genes on a genome-wide scale in , a problematic pathogen for which treatment options are limited. That foundational study used large-scale transposon mutagenesis to identify 404 protein-encoding genes as likely to be essential for vegetative growth of the epidemic strain R20291.
View Article and Find Full Text PDFInvest Ophthalmol Vis Sci
September 2025
Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States.
Purpose: Adeno-associated viruses (AAVs) have become the preferred vector for gene therapy in ophthalmology. However, requirements for specific cell surface receptors limit AAV-mediated retinal cell transduction efficiency. This led to the need to engineer novel AAV vectors for widespread retinal transduction and transgene expression.
View Article and Find Full Text PDFBrain
September 2025
Okinawa Institute of Science and Technology Graduate University, Okinawa, 904-0495, Japan.
Animal models of the pathology of Parkinson's disease (PD) have provided most of the treatments to date, but the disease is restricted to human patients. In vitro models using human pluripotent stem cells (hPSCs)-derived neural organoids have provided improved access to study PD etiology. This study established a method to generate human striatal-midbrain assembloids (hSMAs) from hPSCs for modeling alpha-synuclein (α-syn) propagation and recapitulating basal ganglia circuits, including nigrostriatal and striatonigral pathways.
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