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 sub-classification of circulating tumor cells, a key step towards the identification of true metastasis-initiating cells. Thus, this nano-enabled platform holds great potential for studying the biology of rare tumor cells and for differential diagnosis of cancer progression and metastasis.

Nanobiotechnology for the Therapeutic Targeting of Cancer Cells in Blood

“…5,10,47 To solve this problem, nanomaterials have been developed to enable high-density coatings of different capture molecules such as antibodies for improved sensitivity of CTC detection. 148 Recently, silicon nanopillars, 144 quartz nanowires 81 and TiO 2 nanofibers 159 have been used to trap CTCs, with enhanced capture efficiency due to the higher aspect ratio of the nanomaterials. Moreover, when such nanomaterial-based platforms have been integrated with flowbased systems such as microfluidic devices, a significant increase in capture yield is observed via continuous flow of patient blood through the devices.…”

Morphology of CD4+T Lymphocytes Bound on Nano-Patterened Substrates for Sensing the Size of Nanoholes

“…The QNHA samples are ready to collect incoming cell suspension containing primary mouse CD4 and CD8 T-lymphocytes, natural killer (NK), natural killer T (NKT), and B cells. We used this surface functionalized method to separate targeting specific cells (in our case CD4 + T-lymphocytes) among different kinds of cells via novel STR-biotin conjugation technique to capture the incoming targeting cells in PBS solution as we reported previously [8,9]. More detailed information can be found in previous reports [8,9].…”

“…Furthermore, nanostructures have been increasingly used for studies on cell interaction with solid nanostructures because unique properties of nanostructured surfaces enable a variety of novel functions of immobilized cells on the nanostructured surfaces [4,5]. To date, these nanostructures were prepared by various methods including the polymeric templating method [6], vaporliquid-solid (VLS) growth [7], dry-etching process using nanosphere lithography technique [8,9]. Recently, we have also demonstrated a novel platform for separating CD4 + T lymphocytes from mouse splenocytes using streptavidin (STR)-functionalized and vapor-liquid-solid (VLS)-grown silicon nanowire (SiNW) [7] as well as transparent quartz nanopillar (QNP) arrays [8,9] having a higher separation efficiency of 93-95.3%.…”

“…To date, these nanostructures were prepared by various methods including the polymeric templating method [6], vaporliquid-solid (VLS) growth [7], dry-etching process using nanosphere lithography technique [8,9]. Recently, we have also demonstrated a novel platform for separating CD4 + T lymphocytes from mouse splenocytes using streptavidin (STR)-functionalized and vapor-liquid-solid (VLS)-grown silicon nanowire (SiNW) [7] as well as transparent quartz nanopillar (QNP) arrays [8,9] having a higher separation efficiency of 93-95.3%. However, there is no study on how the various size and shape-matched nanostructures and nanomaterials interact in the primary cells on the nanostructured substrates.…”

Cuprous oxide nanoparticle-inhibited melanoma progress by targeting melanoma stem cells

“…7 Many nano medicines such as gold nanoparticles, metallic multi-segments, silicon nanowire, iron core-gold shell nanoparticles, chitosan nanoparticles, and gold nanorods have been designed for tumor therapy. [8][9][10][11] As a new member of potential antitumor drugs, cuprous oxide nanoparticles (CONPs) have been proven not only to induce apoptosis of tumor cells selectively in vitro but also to inhibit the growth and metastasis of melanoma by targeting mitochondria with little hepatic and renal toxicities in mice. 12,13 In the meantime, a lot of studies focusing on the biological properties of melanoma provided more strategies for finding or designing new drugs and therapies.…”