Single cell magnetic microscopy with multicolor superparamagnetic probes.

具有多色超顺磁探针的单细胞磁显微镜。

• 批准号: 9789307
• 负责人: Victor Marcel Acosta
• 金额: $ 18.34万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789307
• 关键词: Antibodies; BT 474; Biochemical; Breast Cancer Cell; Caliber; Cancer Biology; Cancer Diagnostics; Cells; Chemistry; Color; Contrast Media; Cryoultramicrotomy; Data; Dependence; Detection; Development; Diagnosis; Diamond; Electron Microscopy; Evolution; Film; Filtration; Fluorescence; Goals; Image; Imaging Phantoms; Immunogenetics; In Vitro; Individual; Label; Magnetic Resonance Imaging; Magnetic nanoparticles; Magnetism; Membrane; Methods; Microfluidic Microchips; Microfluidics; Microscope; Microscopy; Monitor; Morphology; Neoplasm Circulating Cells; Non-Invasive Cancer Detection; Nuclear; Physics; Physiologic pulse; Physiological; Population; Property; Quality Control; Radioactivity; Relaxation; Research; Residual state; Resolution; Screening for cancer; Signal Transduction; Sorting - Cell Movement; Specificity; Stains; Subgroup; Surface; Techniques; Testing; Thick; Time; Tissues; base; bioimaging; cancer cell; clinically relevant; experimental study; imaging modality; imaging platform; improved; in vivo; innovation; iron oxide nanoparticle; magnetic field; millimeter; nanoparticle; off-label use; optical imaging; particle; programs; response; scale up; sensor; superparamagnetism; time use; tissue phantom; tumor

Project Summary: ​Single-cell magnetic microscopy with multicolor superparamagnetic probes. The ability to image small tumors or individual circulating tumor cells non-invasively could have a major impact on the early detection of cancer. However, this remains a major technical challenge, as target cells must be abundantly labeled with contrast agents with a high degree of specificity, and ultra-sensitive imaging modalities must be employed that retain sensitivity even when separated by thick tissue. Superparamagnetic iron-oxide nanoparticles (SPIONs) are promising probes for these purposes, as they are relatively non-toxic and magnetic fields are not attenuated by tissue. Our group has recently developed a technique, diamond magnetic microscopy, capable of imaging individual 22-nm SPIONs within ~1 µm of the diamond surface with high spatial resolution (~300 nm), wide field of view (up to several mm), and >1 kHz frame rate. Here we propose a research program to extend the platform to image immunomagnetically labeled cells using multicolor SPION probes and diamond magnetic microscopy. Individual “colors” of SPIONs will be sorted based on their relaxation times using a microfluidic chamber outfitted with miniature magnets. We will use these probes in combination with diamond magnetic microscopy for two proof-of-principle experiments. In the first, two SPION colors will be introduced to breast cancer cells ​in vitro​; one batch will be functionalized with the Her2 antibody and the other will be labeled with isotype-matched non-specific antibody. The cells will be fixed at various timesteps afterwards and imaged using a dual-channel diamond magnetic microscope. The microscope can differentiate SPION “colors” based on their different temporal response to pulsed magnetic fields, revealing images of the sub-cellular distribution of specific and non-specifically-bound SPIONs. These data will be used to unravel the evolution of specific and non-specific internalization dynamics. In the second experiment, we will use a scaled-up version of the diamond magnetic microscope to detect individual magnetically-labeled cells flowing at physiological rates (up to 1 cm/s) through a 1-mm-thick tissue phantom. To accomplish these aims, we have assembled a team with broad strengths, including SPION chemistry, cancer biology, and sensor physics. If successful, the detection platform may eventually be used to detect CTCs and small tumors ​in vivo​.

项目摘要：单细胞磁显微镜与超顺磁性探针。 对小肿瘤或单个循环肿瘤细胞进行非侵入性成像的能力可能具有 对癌症的早期发现有重大影响。然而，这仍然是一个主要的技术问题。 这是一个挑战，因为靶细胞必须用造影剂大量标记， 特异性和超灵敏成像模式，必须采用保持灵敏度， 被厚厚的组织隔开超顺磁性氧化铁纳米颗粒（SPION）是 有前途的探针，为这些目的，因为他们是相对无毒和磁场是 没有被组织削弱。 我们小组最近开发了一种技术，金刚石磁显微镜，能够 对金刚石表面~1 µm范围内的单个22 nm SPION进行成像， 分辨率（~300 nm）、宽视场（高达几mm）和>1 kHz帧速率。这里我们 提出一项研究计划，将该平台扩展到免疫磁性标记细胞的成像 使用磁探针和金刚石磁性显微镜。 SPION的各个“颜色”将根据它们的弛豫时间进行分类， 装有微型磁铁的房间我们将结合使用这些探头， 金刚石磁性显微镜用于两个原理验证实验。在第一，两个SPION 颜色将在体外被引入乳腺癌细胞;一批将被功能化的 Her 2抗体，另一个将用同种型匹配的非特异性抗体标记。的 细胞将被固定在不同的时间步后，并使用双通道钻石成像 磁性显微镜显微镜可以区分SPION“颜色”的基础上，他们的不同 脉冲磁场的时间响应，揭示了亚细胞分布的图像， 特异性和非特异性结合SPION。这些数据将被用来揭示 特异性和非特异性内化动力学。在第二个实验中，我们将使用 放大版的金刚石磁性显微镜，用于检测单个磁性标记 细胞以生理速率（高达1 cm/s）流过1 mm厚的组织模型。 为了实现这些目标，我们组建了一个具有广泛优势的团队，包括SPION 化学、癌症生物学和传感器物理学。如果成功，检测平台可以 最终用于检测体内CTC和小肿瘤。