Mapping Cancer Metabolism by Mid-infrared Photothermal Microscopy

通过中红外光热显微镜绘制癌症代谢图

• 批准号: 10491322
• 负责人: Ji-Xin Cheng
• 金额: $ 38.56万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10491322
• 关键词: Address; Amino Acids; Amplifiers; Area; Biopsy; Caliber; Carbohydrates; Cells; Cellular Metabolic Process; Chemicals; Cholesterol; Cholesterol Esters; Cisplatin; Detection; Development; Digital Signal Processing; Drug resistance; Electronics; Fatty Acids; Fingerprint; Frequencies; Glucose; Goals; Image; Imaging Device; Lateral; Lead; Lighting; Lipids; Lipoproteins; Location; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Maps; Mass Spectrum Analysis; Measurement; Measures; Mediating; Metabolic; Metabolism; Microscope; Microscopy; Molecular; NMR Spectroscopy; Noise; Nucleic Acids; Organism; Pathogenesis; Pattern; Performance; Photons; Physiologic pulse; Play; Proteins; Pump; Raman Spectrum Analysis; Refractive Indices; Renal carcinoma; Research; Resistance; Resolution; Role; Sampling; Scanning; Scheme; Semiconductors; Serum; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Speed; Subcellular structure; System; Technology; Testing; Time; Tissue Extracts; Tissues; Universities; absorption; base; cancer cell; drug development; glucose uptake; high resolution imaging; imaging platform; imaging system; improved; indexing; infrared spectroscopy; innovation; leukemia; lipid biosynthesis; malignant breast neoplasm; malignant mouth neoplasm; metabolic imaging; metal oxide; microscopic imaging; nanoparticle; neoplastic cell; oxidation; programs; refractory cancer; submicron; success; tool; tumor metabolism; uptake; vibration; Address; Amino Acids; Amplifiers; Area; Biopsy; Caliber; Carbohydrates; Cells; Cellular Metabolic Process; Chemicals; Cholesterol; Cholesterol Esters; Cisplatin; Detection; Development; Digital Signal Processing; Drug resistance; Electronics; Fatty Acids; Fingerprint; Frequencies; Glucose; Goals; Image; Imaging Device; Lateral; Lead; Lighting; Lipids; Lipoproteins; Location; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Maps; Mass Spectrum Analysis; Measurement; Measures; Mediating; Metabolic; Metabolism; Microscope; Microscopy; Molecular; NMR Spectroscopy; Noise; Nucleic Acids; Organism; Pathogenesis; Pattern; Performance; Photons; Physiologic pulse; Play; Proteins; Pump; Raman Spectrum Analysis; Refractive Indices; Renal carcinoma; Research; Resistance; Resolution; Role; Sampling; Scanning; Scheme; Semiconductors; Serum; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Speed; Subcellular structure; System; Technology; Testing; Time; Tissue Extracts; Tissues; Universities; absorption; base; cancer cell; drug development; glucose uptake; high resolution imaging; imaging platform; imaging system; improved; indexing; infrared spectroscopy; innovation; leukemia; lipid biosynthesis; malignant breast neoplasm; malignant mouth neoplasm; metabolic imaging; metal oxide; microscopic imaging; nanoparticle; neoplastic cell; oxidation; programs; refractory cancer; submicron; success; tool; tumor metabolism; uptake; vibration

Program Summary While altered cell metabolism is emerging as a hallmark of cancer, there is an unmet need for new tools for quantitation of metabolites. NMR spectroscopy, mass spectrometry, FTIR, and Raman spectroscopy are widely used for molecular detection in tissue extracts or intact tissues. Yet, these tools do not indicate the spatial locations of the analytes inside the cell. We address this unmet need via development of a lock-in free, wide- field mid-infrared photothermal (MIP) microscope. Our technology will enable quantitative vibrational imaging of metabolites in live tumor cells and intact biopsies. In MIP microscopy recently developed in the PI lab (Sci Adv 2016), a visible beam probes the thermal effect (e.g. change of refractive index and thermal expansion) induced by a pulsed infrared beam. The MIP signal is then extracted through a lock-in amplifier. To match the IR/visible illumination area, the PI lab further developed a wide-field MIP microscope in which a complementary metal– oxide–semiconductor (CMOS) camera and synchronization electronics are harnessed for whole-field lock-in detection (Sci Adv 2019). Despite these initial successes, the sensitivity of MIP microscopy is limited by the detection schemes. First, the golden standard lock-in detection misses all the harmonic frequencies in the MIP signal. Second, the well-depth of a typical CMOS camera seriously limits the probe power to 0.01 mW at sample. Thus, many averages are needed to reach a reasonable signal to noise ratio. We overcome these difficulties through two innovations. The first one is to digitize the probe photons received by a fast photodiode. Then, in the frequency domain, a match filter is used to extract all MIP signals at fundamental and harmonic frequencies. The second one is to perform patterned probe illumination and collect photons with a photodiode which has a saturation threshold of tens of mW. Then, a MIP image is recovered by matrix inversion. In this “single-pixel camera” approach, the probe power can be increased by 1000 times, which indicates that the speed can be improved 30 times to reach the same signal to noise ratio of wide field MIP at the shot noise limit. The goal of this R33 proposal is to develop a digital signal processing, single pixel camera MIP microscope and validate its potential for high-content cancer metabolic imaging. In particular, we aim to validate a metabolic switch from glucose-mediated lipogenesis to fatty acids uptake/oxidation in ovarian cancers that become resistant to cisplatin. By accomplishing the proposed studies, we will generate a high-speed hyperspectral mid-infrared photothermal chemical imaging platform that is able to map the live cell metabolism at sub-micron spatial resolution. Metabolic imaging of live drug-resistant cancer cells by this platform opens new opportunities of unveiling hidden signatures that can potentially lead to adaptive therapies that inhibit the development of drug resistance in cancers.

程序摘要 虽然改变的细胞代谢正在成为癌症的标志，但对新工具的需求未满足 代谢物的定量。 NMR光谱，质谱法，FTIR和拉曼光谱广泛是广泛的 用于组织提取物或完整组织中的分子检测。但是，这些工具并未表示空间 细胞内分析物的位置。我们通过开发免费，广泛的锁定来解决这种未满足的需求 场中红外光热（MIP）显微镜。我们的技术将实现定量振动成像 活肿瘤细胞和完整活检中的代谢产物。在最近在PI实验室中开发的MIP显微镜中（Sci Adv 2016年），可见的光束探测热效应（例如，折射率和热膨胀的变化）诱导 通过脉冲红外光束。然后通过锁定放大器提取MIP信号。匹配IR/可见 Pi Lab Illumination区域进一步开发了宽场MIP显微镜，其中完整的金属 - 氧化物 - 血管导向器（CMOS）摄像头和同步电子设备用于全场锁定 检测（SCI ADV 2019）。尽管取得了这些最初的成功，但MIP显微镜的灵敏度受到 检测方案。首先，黄金标准锁定检测错过了MIP中的所有谐波频率 信号。其次，典型的CMOS摄像机的良好深度将探针功率严重限制为样品时的0.01兆瓦。 这是需要许多平均值来达到合理的信号与噪声比。我们克服了这些困难 通过两项创新。第一个是将快速光电二极管收到的探针照片数字化。然后，在 频域，匹配过滤器用于在基本和谐波频率下提取所有MIP信号。 第二个是执行模式的探针照明，并用具有一个具有光电二极管的照片 数十MW的饱和阈值。然后，通过矩阵反转恢复MIP图像。在这个“单像素”中 相机”方法，探针功率可以提高1000倍，这表明速度可以是 在射击噪声限制下，改进了30次，达到宽场MIP的相同信号与噪声比。目标 该R33建议是开发数字信号处理，单像素摄像头MIP显微镜并验证其 高含量癌症代谢成像的潜力。特别是，我们旨在验证从 葡萄糖介导的脂肪生成对卵巢癌的脂肪酸摄取/氧化，对顺铂具有抗性。 通过完成拟议的研究，我们将产生高速高光谱中红外光热 化学成像平台能够以亚微米空间分辨率绘制活细胞代谢。代谢 该平台对现场耐药性癌细胞进行成像开辟了新的机会，以揭示隐藏的签名 这可能会导致自适应疗法抑制癌症耐药性的发展。