Photoreceptor loss does not recruit neutrophils despite strong microglial activation
Data files
Aug 07, 2025 version files 9.12 GB
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Lesion_paper_data.zip
9.12 GB
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README.md
1.64 KB
Abstract
In response to central nervous system (CNS) injury, tissue resident immune cells such as microglia and circulating systemic neutrophils are often first responders. The degree to which these cells interact in response to CNS damage is poorly understood, and even less so, in the neural retina which poses a challenge for high resolution imaging in vivo. In this study, we deploy fluorescence adaptive optics scanning light ophthalmoscopy (AOSLO) to study microglia and neutrophils in mice. We simultaneously track immune cell dynamics using label-free phase-contrast AOSLO at micron-level resolution. Retinal lesions were induced with 488 nm light focused onto photoreceptor (PR) outer segments. These lesions focally ablated PRs, with minimal collateral damage to cells above and below the plane of focus. We used in vivo (AOSLO, SLO and OCT) imaging to reveal the natural history of the microglial and neutrophil response from minutes-to-months after injury. While microglia showed dynamic and progressive immune response with cells migrating into the injury locus within 1-day after injury, neutrophils were not recruited despite close proximity to vessels carrying neutrophils only microns away. Post-mortem confocal microscopy confirmed in vivo findings. This work illustrates that microglial activation does not recruit neutrophils in response to acute, focal loss of PRs, a condition encountered in many retinal diseases.
Dataset DOI: 10.5061/dryad.w3r228143
Description of the data and file structure
Data files within are primarily videos and images acquired for a paper "Photoreceptor loss does not recruit neutrophils despite strong microglial activation" submitted to eLife.
Files and variables
File: Lesion_paper_data.zip
Description: 3 sub-folders contain data acquired using Adaptive optics scanning light ophthalmoscopy (AOSLO or AO for short), a scanning laser ophthalmoscope (Heidelberg Spectralis, or HRA), and ex-vivo confocal microscopy. Within each sub-folder, as follows: AOdata-->mouse strain and other label-->acquisition modality (confocal or phase-contrast, this folder is not always present)-->mouse # and date of acquisition. exVivoConfocaldata-->mouse strain and other label for histology-->mouse # and date of acquisition. HRAdata-->mouse # and acquisition data. For folders with a # and a date, the first five digits signify a particular mouse that was imaged and the remaining digits indicate the date the data was acquired. Each imaging session has a manually-recorded note sheet; these PDFs are provided as supplemental information via Zenodo. Note that raw AO videos were quite large (the entire dataset was 3TB), and only data relevant for this paper is included.
Code/software
Most videos and images are in .avi and .tif format and can be opened by many common media software. The ND2 files can be opened with ImageJ by choosing "load as hyperstack". All files can be viewed with ImageJ.