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Optofluidic device for label-free cell classification from whole blood.

ABSTRACT: We demonstrated a unique optofluidic lab-on-a-chip device that can measure optically encoded forward scattering signals. From the design of the spatial pattern, we can measure the position and velocity of each cell in the flow and generate a 2-D cell distribution plot over the cross section of the channel. Moreover, we have demonstrated that the cell distribution is highly sensitive to its size and stiffness. The latter is an important biomarker for cell classification and our method offers a simple and unequivocal method to classify cells by their size and stiffness. We have proved the concept using live and fixed HeLa cells. Due to the stiffness and size difference of neutrophils compared to other types of white blood cells, we have demonstrated detection of neutrophils from other blood cells. Finally, we have performed the test using 5 ?L of human blood. In a greatly simplified blood preparation process, skipping the usual steps of anticoagulation, centrifuge, antibody labelling or staining, filtering, etc., we have demonstrated that our device and detection principle can count neutrophils in whole human blood. Our system is compact, inexpensive and simple to fabricate and operate, having a commodity laser diode and a Si PIN photoreceiver as the main pieces of hardware. Although the results are still preliminary, the studies indicate that this optofluidic device holds promise to be a point-of-care and home care device to measure neutrophil concentration, which is the key indicator of the immune functions for cancer patients undergoing chemotherapy.

SUBMITTER: Wu TF

PROVIDER: S-EPMC3472004 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Optofluidic device for label-free cell classification from whole blood.

Wu Tsung-Feng TF Mei Zhe Z Lo Yu-Hwa YH

Lab on a chip 20121001 19

We demonstrated a unique optofluidic lab-on-a-chip device that can measure optically encoded forward scattering signals. From the design of the spatial pattern, we can measure the position and velocity of each cell in the flow and generate a 2-D cell distribution plot over the cross section of the channel. Moreover, we have demonstrated that the cell distribution is highly sensitive to its size and stiffness. The latter is an important biomarker for cell classification and our method offers a si ...[more]

PMID: 22875178

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Project description:Cancer cells metastasize through the bloodstream either as single migratory circulating tumor cells (CTCs) or as multicellular groupings (CTC-clusters). Existing technologies for CTC enrichment are designed primarily to isolate single CTCs, and while CTC-clusters are detectable in some cases, their true prevalence and significance remain to be determined. Here, we developed a microchip technology (Cluster-Chip) specifically designed to capture CTC-clusters independent of tumor-specific markers from unprocessed blood. CTC-clusters are isolated through specialized bifurcating traps under low shear-stress conditions that preserve their integrity and even two-cell clusters are captured efficiently. Highly parallel architecture of the chip allows deterministic screening of clinically relevant volumes of blood samples at slow, and hence, non-damaging flow rates. Using the Cluster-Chip, we identify CTC-clusters in 30-40% of patients with metastatic cancers of the breast, prostate and melanoma. RNA sequencing of CTC-clusters confirms their tumor origin and identifies leukocytes within the clusters as being tissue-derived macrophages. Together, the development of a device for efficient capture of CTC-clusters will enable detailed characterization of their biological properties and role in cancer metastasis. We used the Cluster-Chip to capture CTC-clusters from the blood of a breast cancer patient with high CTC counts, released CTC-clusters in solution, stained them with TexasRed-conjugated antibodies against the leukocyte cell surface markers CD45, CD14 and CD16, and then isolated intact CTC-clusters individually using a micromanipulator. From a single time point, we retrieved 15 CTC-clusters, and each of those clusters was individually subjected to RNA-sequencing analysis using a next generation platform (SOLiD 5500). In addition, two leukocytes were isolated from the blood of a healthy donor were individually subjected to RNA-sequencing analysis using the same platform.

Dataset Information

Optofluidic device for label-free cell classification from whole blood.

Publications

Optofluidic device for label-free cell classification from whole blood.

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