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Microparticle-tagged image-based cell counting (ImmunoSpin) for CD4 + T cells.

ABSTRACT: Affordable point-of-care (POC) CD4 + T lymphocyte counting techniques have been developed as alternatives to flow cytometry-based instruments caring for patients with human immunodeficiency virus (HIV)-1. However, POC CD4 enumeration technologies can be inaccurate. Here, we developed a microparticle-based visual detector of CD4 + T lymphocytes (ImmunoSpin) using microparticles conjugated with anti-CD4 antibodies, independent of microfluidic or fluorescence detection systems. Visual enumeration of CD4 + T cells under conventional light microscope was accurate compared to flow cytometry. Microparticle-tagged CD4 + T cells were well-recognized under a light microscope. ImmunoSpin showed very good precision (coefficients of variation of ImmunoSpin were ≤ 10%) and high correlation with clinical-grade flow cytometry for the enumeration of CD4 + T cells (y = 0.4232 + 0.9485 × for the %CD4 + T cell count, R² = 0.99). At thresholds of 200 and 350 cells/µL, there was no misclassification of the ImmunoSpin system compared to the reference flow cytometry. ImmunoSpin showed clear differential classification of CD4 + T lymphocytes from granulocytes and monocytes. Because non-fluorescence microparticle-tags and cytospin slides are used in ImmunoSpin, they can be applied to an automatic digital image analyzer. Slide preparation allows long-term storage, no analysis time limitations, and image transfer in remote areas.

SUBMITTER: Hwang SH

PROVIDER: S-EPMC8616869 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:To investigate the feasibility of using an image-based method to correct for distortions induced by various artifacts in the x-ray spectrum recorded with photon-counting detectors for their application in breast computed tomography (CT).The polyenergetic incident spectrum was simulated with the tungsten anode spectral model using the interpolating polynomials (TASMIP) code and carefully calibrated to match the x-ray tube in this study. Experiments were performed on a Cadmium-Zinc-Telluride (CZT) photon-counting detector with five energy thresholds. Energy bins were adjusted to evenly distribute the recorded counts above the noise floor. BR12 phantoms of various thicknesses were used for calibration. A nonlinear function was selected to fit the count correlation between the simulated and the measured spectra in the calibration process. To evaluate the proposed spectral distortion correction method, an empirical fitting derived from the calibration process was applied on the raw images recorded for polymethyl methacrylate (PMMA) phantoms of 8.7, 48.8, and 100.0 mm. Both the corrected counts and the effective attenuation coefficient were compared to the simulated values for each of the five energy bins. The feasibility of applying the proposed method to quantitative material decomposition was tested using a dual-energy imaging technique with a three-material phantom that consisted of water, lipid, and protein. The performance of the spectral distortion correction method was quantified using the relative root-mean-square (RMS) error with respect to the expected values from simulations or areal analysis of the decomposition phantom.The implementation of the proposed method reduced the relative RMS error of the output counts in the five energy bins with respect to the simulated incident counts from 23.0%, 33.0%, and 54.0% to 1.2%, 1.8%, and 7.7% for 8.7, 48.8, and 100.0 mm PMMA phantoms, respectively. The accuracy of the effective attenuation coefficient of PMMA estimate was also improved with the proposed spectral distortion correction. Finally, the relative RMS error of water, lipid, and protein decompositions in dual-energy imaging was significantly reduced from 53.4% to 6.8% after correction was applied.The study demonstrated that dramatic distortions in the recorded raw image yielded from a photon-counting detector could be expected, which presents great challenges for applying the quantitative material decomposition method in spectral CT. The proposed semi-empirical correction method can effectively reduce these errors caused by various artifacts, including pulse pileup and charge sharing effects. Furthermore, rather than detector-specific simulation packages, the method requires a relatively simple calibration process and knowledge about the incident spectrum. Therefore, it may be used as a generalized procedure for the spectral distortion correction of different photon-counting detectors in clinical breast CT systems.

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Microparticle-tagged image-based cell counting (ImmunoSpin) for CD4 + T cells.

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