Cellular & Molecular Imaging

Fluorescence imaging across the near-infrared (NIR, 700–1000 nm) and shortwave infrared (SWIR, 1000–2000 nm) regions offers significant advantages for biomedical applications, yet photophysical enhancements achieved with NIR and SWIR chromophores observed in solution often fail to translate to complex biological environments. Fluorous-soluble fluorophores, fluorofluorophores, face additional challenges, exhibiting poor brightness and photostability when dissolved in perfluorocarbons (PFCs) due to unfavorable interactions with the fluorous phase. Here, we report counterion exchange as a strategy to enhance the photophysical properties of two heptamethine cyanine fluorofluorophore for NIR and SWIR imaging. Exchanging the small chloride counterion with a large, fluorinated aryl borate counterions significantly improved the brightness (10-fold) and photostability (57-fold) in PFCs. These enhancements were successfully translated across multiple biological systems from macrophage cells to NIR imaging zebrafish retinal tissue and finally to SWIR imaging in mice. These results demonstrate that strategic counterion modification provides a straightforward approach to optimize fluorofluorophores, with solution-phase improvements that translate to in vivo NIR and SWIR imaging.

The Darwin Tree of Life project has the goal to sequence the genomes of 70,000 species of eukaryotic organisms in Britain and Ireland. This is a collection of photographs of the samples included in the study, provided by the National History Museum (NHM).

Lepidoptera, i.e. butterflies and moths, are vital components of the global ecosystem. Project Psyche is a scientific research project established to sequence the genomes of all butterflies and moths of Europe; helping to conserve, protect and drive innovation.

The Darwin Tree of Life project has the goal to sequence the genomes of 70,000 species of eukaryotic organisms in Britain and Ireland. This is a collection of photographs of the samples included in the study, provided by the National History Museum (NHM).

OME-NGFF converted study from idr0090. Drug resistance threatens the effective prevention and treatment of an ever-increasing range of human infections. This highlights an urgent need for new and improved drugs with novel mechanisms of action to avoid cross-resistance. Current cell-based drug screens are, however, restricted to binary live/dead readouts with no provision for mechanism of action prediction. Machine learning methods are increasingly being used to improve information extraction from imaging data. Such methods, however, work poorly with heterogeneous cellular phenotypes and generally require time-consuming human-led training. We have developed a semi-supervised machine learning approach, combining human- and machine-labelled training data from mixed human malaria parasite cultures. Designed for high-throughput and high-resolution screening, our semi-supervised approach is robust to natural parasite morphological heterogeneity and correctly orders parasite developmental stages. Our approach also reproducibly detects and clusters drug-induced morphological outliers by mechanism of action, demonstrating the potential power of machine learning for accelerating cell-based drug discovery.

DNA-FISH and BRD4 immunofluorescence confocal stacks targeting PTC regions described in Klonizakis et al. 2023

Immunofluorescence microscopy images supporting expression of HA-tagged PfMD3 in specific stages of P. falciparum development

Raw data corresponding to the publication, "Cell surface localisation of GPI-anchored receptors in Trypanosoma brucei", published in eLife (https://doi.org/10.7554/eLife.107191.1)

Trypanosoma brucei bloodstream form tagging protein localisation data. Widefield epifluorescence microscope images of protein subcellular localisation in the bloodstream form life cycle stage of the unicellular eukaryotic pathogen Trypanosoma brucei by endogenous tagging with mNeonGreen (mNG). This deposition includes a summary of the localisations, primer sequences, provided in a directory structure analogous to the TrypTag genome-wide procyclic form project: https://doi.org/10.6019/S-BIAD1866 .

Raw data corresponding to the publication "Continuous endosomes form functional subdomains and orchestrate rapid membrane trafficking in trypanosomes" published in eLife (https://doi.org/10.7554/eLife.91194.2).