DOI: 10.5061/dryad.w3r228143
title: Photoreceptor loss does not recruit neutrophils despite strong microglial activation

description: 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.
description: # Photoreceptor loss does not recruit neutrophils despite strong
 microglial activation Dataset DOI:
 [10.5061/dryad.w3r228143](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:  AO*data--&gt;mouse strain and other
 label--&gt;acquisition modality (confocal or phase-contrast, this folder
 is not always present)--&gt;mouse # and date of acquisition.
 exVivo*Confocal*data--&gt;mouse strain and other label for
 histology--&gt;mouse # and date of acquisition. HRA*data--&gt;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.