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Nanowire substrate-based laser scanning cytometry for quantitation of circulating tumor cells.

ABSTRACT: We report on the development of a nanowire substrate-enabled laser scanning imaging cytometry for rare cell analysis in order to achieve quantitative, automated, and functional evaluation of circulating tumor cells. Immuno-functionalized nanowire arrays have been demonstrated as a superior material to capture rare cells from heterogeneous cell populations. The laser scanning cytometry method enables large-area, automated quantitation of captured cells and rapid evaluation of functional cellular parameters (e.g., size, shape, and signaling protein) at the single-cell level. This integrated platform was first tested for capture and quantitation of human lung carcinoma cells from a mixture of tumor cells and leukocytes. We further applied it to the analysis of rare tumor cells spiked in fresh human whole blood (several cells per mL) that emulate metastatic cancer patient blood and demonstrated the potential of this technology for analyzing circulating tumor cells in the clinical settings. Using a high-content image analysis algorithm, cellular morphometric parameters and fluorescence intensities can be rapidly quantitated in an automated, unbiased, and standardized manner. Together, this approach enables informative characterization of captured cells in situ and potentially allows for subclassification of circulating tumor cells, a key step toward the identification of true metastasis-initiating cells. Thus, this nanoenabled platform holds great potential for studying the biology of rare tumor cells and for differential diagnosis of cancer progression and metastasis.

SUBMITTER: Lee SK

PROVIDER: S-EPMC3381426 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature

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Nanowire substrate-based laser scanning cytometry for quantitation of circulating tumor cells.

Lee Sang-Kwon SK Kim Gil-Sung GS Wu Yu Y Kim Dong-Joo DJ Lu Yao Y Kwak Minsuk M Han Lin L Hyung Jung-Hwan JH Seol Jin-Kyeong JK Sander Chantal C Gonzalez Anjelica A Li Jie J Fan Rong R

Nano letters 20120531 6

We report on the development of a nanowire substrate-enabled laser scanning imaging cytometry for rare cell analysis in order to achieve quantitative, automated, and functional evaluation of circulating tumor cells. Immuno-functionalized nanowire arrays have been demonstrated as a superior material to capture rare cells from heterogeneous cell populations. The laser scanning cytometry method enables large-area, automated quantitation of captured cells and rapid evaluation of functional cellular ...[more]

PMID: 22646476

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Project description:To develop and test the application of an adaptive optics scanning laser ophthalmoscope (AOSLO) with eye tracking for high-resolution microperimetric testing.An AOSLO was used to conduct simultaneous high-resolution retinal imaging and visual function testing in six normal subjects. Visual sensitivity was measured at test locations between the fovea and 5.0° eccentricity via an increment threshold approach using a 40-trial, yes-no adaptive Bayesian staircase procedure (QUEST). A high-speed eye tracking algorithm enabled real-time video stabilization and the delivery of diffraction-limited Goldmann I-sized stimuli (diameter = 6.5 arc min = ?32 ?m; ? = 680 nm) to targeted retinal loci for 200 ms. Test locations were selected either manually by the examiner or automatically using Fourier-based image registration. Cone spacing was assessed at each test location and sensitivity was plotted against retinal eccentricity. Finally, a 4.2 arc min stimulus was used to probe the angioscotoma associated with a blood vessel located at 2.5° eccentricity.Visual sensitivity decreases with eccentricity at a rate of -1.32 dB/deg (R = 0.60). The vertical and horizontal errors of the targeted stimulus delivery algorithm averaged 0.81 and 0.89 arc min (?4 ?m), respectively. Based on a predetermined exclusion criterion, the stimulus was successfully delivered to its targeted location in 90.1% of all trials. Automated recovery of test locations afforded the repeat testing of the same set of cones over a period of 3 months. Thresholds measured over a parafoveal blood vessel were 1.96 times higher (p < 0.05; one-tailed t-test) than those measured in directly adjacent retina.AOSLO-based microperimetry has the potential to test visual sensitivity with fine retinotopic precision. Automated recovery of previously tested locations allows these measures to be tracked longitudinally. This approach can be implemented by researchers interested in establishing the functional correlates of photoreceptor mosaic structure in patients with retinal disease.

Dataset Information

Nanowire substrate-based laser scanning cytometry for quantitation of circulating tumor cells.

Publications

Nanowire substrate-based laser scanning cytometry for quantitation of circulating tumor cells.

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