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In vivo quantitative analysis of anterior chamber white blood cell mixture composition using spectroscopic optical
Ruobing Qian1, Ryan P McNabb2, Kevin C Zhou1
1Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
Insights
Spectroscopic optical coherence tomography (SOCT) can now estimate white blood cell (WBC) composition in the anterior chamber. This non-invasive method shows promise for diagnosing intraocular inflammation like uveitis.
Area of Science:
- Ophthalmology
- Biomedical Optics
- Cellular Biology
Background:
- Anterior uveitis, a common eye inflammation, is diagnosed by detecting white blood cells (WBCs) in the anterior chamber (AC).
- Current methods like AC paracentesis are invasive, requiring needle insertion to analyze AC fluid.
- Previous work established spectroscopic optical coherence tomography (SOCT) for differentiating blood cells (RBCs, WBC subtypes) in vitro and in excised eyes.
Purpose of the Study:
- To develop and validate a non-invasive SOCT method for estimating the composition of mixed blood cells in the anterior chamber.
- To assess the accuracy of this SOCT method for in vitro and in vivo applications.
Main Methods:
- Utilized SOCT to analyze backscattered light spectra from individual cells, applying Mie theory to determine size distributions.
- Developed a method to estimate cell mixture composition by fitting observed SOCT distributions with weighted combinations of reference size distributions (WBC subtypes) derived from kernel density estimation.
- Validated the method in vitro by comparing SOCT-derived concentrations with hemocytometer counts and pre-mixing SOCT images.
Main Results:
- Demonstrated accurate estimation of in vitro blood cell mixtures.
- A pilot in vivo study showed congruence between the SOCT method and AC paracentesis in two uveitis patients.
- SOCT successfully differentiated between granulocytes, lymphocytes, and monocytes based on their characteristic size distributions.
Conclusions:
- The developed SOCT-based method offers a promising non-invasive approach for quantifying cellular composition in the anterior chamber.
- This technique has the potential to provide crucial diagnostic information for conditions like uveitis.
- Further validation is warranted for broader clinical application.
Abstract:
Anterior uveitis is the most common form of intraocular inflammation, and one of its main signs is the presence of white blood cells (WBCs) in the anterior chamber (AC). Clinically, the true composition of cells can currently only be obtained using AC paracentesis, an invasive procedure to obtain AC fluid requiring needle insertion into the AC. We previously developed a spectroscopic optical coherence tomography (SOCT) analysis method to differentiate between populations of RBCs and subtypes of WBCs, including granulocytes, lymphocytes and monocytes, both in vitro and in ACs of excised porcine eyes. We have shown that different types of WBCs have distinct characteristic size distributions, extracted from the backscattered reflectance spectrum of individual cells using Mie theory. Here, we further develop our method to estimate the composition of blood cell mixtures, both in vitro and in vivo. To do so, we estimate the size distribution of unknown cell mixtures by fitting the distribution observed using SOCT with a weighted combination of reference size distributions of each WBC type calculated using kernel density estimation. We validate the accuracy of our estimation in an in vitro study, by comparing our results for a given WBC sample mixture with the cellular concentrations measured by a hemocytometer and SOCT images before mixing. We also conducted a small in vivo quantitative cell mixture validation pilot study which demonstrates congruence between our method and AC paracentesis in two patients with uveitis. The SOCT based method appears promising to provide quantitative diagnostic information of cellular responses in the ACs of patients with uveitis.
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