Label free measurement of retinal blood cell flux, velocity, hematocrit and capillary width in the living mouse eye

2016/09/23 by Andrés Guevara–Torres, Aby Joseph, Jesse Schallek · 1 voice · 28 citations
Medicine · Biochemistry, Genetics and Molecular Biology · #Glaucoma and retinal disorders #Retinal Diseases and Treatments #Advanced Fluorescence Microscopy Techniques

paper · doi:10.1364/boe.7.004228

Abstract

Measuring blood cell dynamics within the capillaries of the living eye provides crucial information regarding the health of the microvascular network. To date, the study of single blood cell movement in this network has been obscured by optical aberrations, hindered by weak optical contrast, and often required injection of exogenous fluorescent dyes to perform measurements. Here we present a new strategy to non-invasively image single blood cells in the living mouse eye without contrast agents. Eye aberrations were corrected with an adaptive optics camera coupled with a fast, 15 kHz scanned beam orthogonal to a capillary of interest. Blood cells were imaged as they flowed past a near infrared imaging beam to which the eye is relatively insensitive. Optical contrast of cells was optimized using differential scatter of blood cells in the split-detector imaging configuration. Combined, these strategies provide label-free, non-invasive imaging of blood cells in the retina as they travel in single file in capillaries, enabling determination of cell flux, morphology, class, velocity, and rheology at the single cell level.

Citations

• Identification of Distinct Luminal Domains for Macromolecules, Erythrocytes, and Leukocytes Within Mammalian Capillaries

Cited by

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• High-speed adaptive optics line scan confocal retinal imaging for human eye. [europepmc]
• Capillary red blood cell velocimetry by phase-resolved optical coherence tomography. [europepmc]
• Spatio-temporal dynamics of cerebral capillary segments with stalling red blood cells. [europepmc]
• Noninvasive in vivo characterization of erythrocyte motion in human retinal capillaries using high-speed adaptive optics near-confocal imaging. [europepmc]
• Versatile multi-detector scheme for adaptive optics scanning laser ophthalmoscopy. [europepmc]
• Imaging single-cell blood flow in the smallest to largest vessels in the living retina. [europepmc]
• Mapping flow velocity in the human retinal capillary network with pixel intensity cross correlation. [europepmc]
• Cellular Control of Brain Capillary Blood Flow: In Vivo Imaging Veritas. [europepmc]
• Near infrared adaptive optics flood illumination retinal angiography. [europepmc]
• Imaging Retinal Activity in the Living Eye. [europepmc]
• Imaging relative stasis of the blood column in human retinal capillaries. [europepmc]
• Origin of cell contrast in offset aperture adaptive optics ophthalmoscopy. [europepmc]
• In vivo imaging of corneal nerves and cellular structures in mice with Gabor-domain optical coherence microscopy. [europepmc]
• Label-free imaging of immune cell dynamics in the living retina using adaptive optics. [europepmc]
• Intravital Imaging of Circulating Red Blood Cells in the Retinal Vasculature of Growing Mice. [europepmc]
• Association between erythrocyte dynamics and vessel remodelling in developmental vascular networks. [europepmc]
• Development of a Preclinical Laser Speckle Contrast Imaging Instrument for Assessing Systemic and Retinal Vascular Function in Small Rodents. [europepmc]
• Imaging the dynamics of individual processes of microglia in the living retina in vivo . [europepmc]
• Computational investigation of blood cell transport in retinal microaneurysms. [europepmc]
• In Vivo Capillary Structure and Blood Cell Flux in the Normal and Diabetic Mouse Eye. [europepmc]
• Adaptive optics for high-resolution imaging. [europepmc]
• Mechanical regulation of the early stages of angiogenesis. [europepmc]
• Methods to measure blood flow and vascular reactivity in the retina. [europepmc]
• Evolution of adaptive optics retinal imaging [Invited]. [europepmc]
• Changes to the shape, orientation and packing of red cells as a function of retinal capillary size. [europepmc]

Discussions

• If you scan a point of light fast enough on the retina👁️, you can image blood cells ONE-BY-ONE as they cross the beam Our lab is developing tech called "in vivo flow cytometry" @flaumeye.bsky.so [bsky, 15 points, 3 comments]

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