Bentley-Abbot, Calum and Heslop, Rhiannon and Pirillo, Chiara and Chandrasegaran, Praveena and McConnell, Gail and Roberts, Ed and Hutchinson, Edward and MacLeod, Annette (2024) An easy to use tool for the analysis of subcellular mRNA transcript colocalisation in smFISH data. Scientific Reports, 14: 8348. ISSN 2045-2322
AI Summary:
FISHtoFigure is a new software tool developed specifically for the analysis of mRNA abundance and co-expression in QuPath-quantified, multi-labelled smFISH data.AI Topics:
Single molecule fluorescence in situ hybridisation (smFISH) has become a valuable tool to investigate the mRNA expression of single cells. However, it requires a considerable amount of programming expertise to use currently available open-source analytical software packages to extract and analyse quantitative data about transcript expression. Here, we present FISHtoFigure, a new software tool developed specifically for the analysis of mRNA abundance and co-expression in QuPath-quantified, multi-labelled smFISH data. FISHtoFigure facilitates the automated spatial analysis of transcripts of interest, allowing users to analyse populations of cells positive for specific combinations of mRNA targets without the need for computational image analysis expertise. As a proof of concept and to demonstrate the capabilities of this new research tool, we have validated FISHtoFigure in multiple biological systems. We used FISHtoFigure to identify an upregulation in the expression of Cd4 by T-cells in the spleens of mice infected with influenza A virus, before analysing more complex data showing crosstalk between microglia and regulatory B-cells in the brains of mice infected with Trypanosoma brucei brucei. These analyses demonstrate the ease of analysing cell expression profiles using FISHtoFigure and the value of this new tool in the field of smFISH data analysis.
Title | An easy to use tool for the analysis of subcellular mRNA transcript colocalisation in smFISH data |
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Creators | Bentley-Abbot, Calum and Heslop, Rhiannon and Pirillo, Chiara and Chandrasegaran, Praveena and McConnell, Gail and Roberts, Ed and Hutchinson, Edward and MacLeod, Annette |
Identification Number | 10.1038/s41598-024-58641-3 |
Date | 9 April 2024 |
Divisions | College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine College of Medical Veterinary and Life Sciences > School of Cancer Sciences College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research |
Publisher | Nature Research |
Additional Information | The work to produce this dataset was funded by a Sir Henry Wellcome postdoctoral fellowship (221640/Z/20/Z to J.F.Q.) and a Wellcome Trust ISSF Catalyst grant awarded to J.F.Q. (204820/Z/16/Z to JFQ). We also thank Colin Loney for his assistance in the acquisition of the images forming the spleen dataset used during validation. We also thank Ruaridh Wilson for providing feedback from the perspective of a computational scientist during the writing of this manuscript. |
URI | https://pub.demo35.eprints-hosting.org/id/eprint/297 |
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Item Type | Article |
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Depositing User | Unnamed user with email ejo1f20@soton.ac.uk |
SWORD Depositor | Users 37347 not found. |
Date Deposited | 11 Jun 2025 16:36 |
Revision | 22 |
Last Modified | 12 Jun 2025 11:17 |
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