Circularly Polarised Luminescence Laser Scanning Confocal Microscopy

圆偏振发光激光扫描共焦显微镜

• 批准号: BB/X001172/1
• 负责人: Robert Pal
• 金额: $ 57.28万
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX001172%2F1
• 关键词: Circularly; Polarised; Luminescence; Laser; Scanning

Marginal organelle specific intra-cellular temperature changes often signal the early onset of life-altering diseases, such as cancer or acute mitochondrial disorder. These currently cannot be monitored and there is also no optical microscopy technique that enables the bio-imaging community to study chiral molecular interactions based on chiroptical (chiral optical) activity. With our unique expertise, we seek to address this by combining lanthanide coordination chemistry and a versatile new instrumental design. Once fully developed and validated it could open new horizons towards disease progression studies.Temperature variations occur in cell division, gene expression, enzymatic reactions and natural cell metabolism. They may also signal the onset of pathological states and dysfunctions. Cancer tissue typically has a higher temperature (T) than surrounding healthy tissue, associated with higher cellular metabolic rates. The accurate monitoring of intracellular temperature may enable a better understanding of complex cellular events and aid the detection of diseases at the cellular level, allowing new methods of early diagnosis and therapy to be developed. Temperature monitoring is also of fundamental relevance in thermal therapies, such as hyperthermia and thermal ablation, explored as minimally invasive alternatives to surgical procedures. The proposed project will harness the phenomenon of Circularly Polarised Luminescence (CPL), where different enantiomers of the same chemical entity produce different handedness (left or right) of light. Luminescent lanthanide complexes have been shown to possess a remarkably unique photophysical property, where the ratio of their left- and right-handed luminescence is extremely sensitive to the temperature of their surroundings. Therefore, these could be exploited as high precision intracellular temperature probes.Our newly developed all solid-state CPL spectrometer is a paradigm shift in CPL spectroscopy, and due to its small size and versatility, it enables truly widespread application. Adapting this technique into a high spatial (optical precision) and temporal (acquisition speed) resolution microscope setup is straightforward due to our expertise and track record in instrument development. Therefore, we propose constructing and validating the world's first Confocal Laser Scanning CPL Microscope (CPL-LSCM). We are fully aware that deep tissue imaging using conventional optical microscopy is challenging. Hence, we also plan to incorporate low energy, biologically safe Near Infra-Red (NIR) multiphoton activation (MP) to increase the observable depth of tissue. It will enable unprecedented live-cell enantioselective chiroptical microscopy and provides immense scientific value, opening new horizons for studying and tracking emissive chiral molecules, both endogenous and engineered bio-probes.With our pioneering work in rapid CPL spectroscopy it is within our grasp to achieve. Our proposal has great potential beyond the benefits associated with the wide and diverse multidisciplinary scientific community. It could also generate immense commercial interest, initiate a new chapter in modern-day live-cell optical microscopy, and shed light on the previously unexplored biochemical processes that fundamentally underpin life and life-threatening disease progressions.Our overarching aim is to establish it as the go to 'research tool' in helping answer fundamental questions, such as why life has been founded on one particular (L) enantiomer of its chiral amino acid building blocks but needs the opposite (D) enantiomer of glucose. It can also expand our understanding of complex bio-molecules such as enzymes and elementary biochemical process such cell division, unearthing the explicit role and driving force of chirality in them.

边缘细胞器特异性细胞内温度变化通常是改变生命的疾病（如癌症或急性线粒体疾病）早期发作的信号。这些目前无法监测，也没有光学显微镜技术，使生物成像界研究手性分子相互作用的基础上手性光学（手性光学）活动。凭借我们独特的专业知识，我们试图通过结合镧系元素配位化学和多功能的新仪器设计来解决这个问题。一旦完全开发和验证，它将为疾病进展研究开辟新的视野。温度变化发生在细胞分裂，基因表达，酶促反应和自然细胞代谢中。它们也可能预示着病理状态和功能障碍的开始。癌症组织通常具有比周围健康组织更高的温度（T），与更高的细胞代谢率相关。细胞内温度的准确监测可以更好地理解复杂的细胞事件，并有助于在细胞水平上检测疾病，从而开发早期诊断和治疗的新方法。温度监测在热疗法中也具有基本相关性，例如热疗和热消融，被探索为外科手术的微创替代方案。拟议的项目将利用圆偏振发光（CPL）现象，其中相同化学实体的不同对映体产生不同的光旋向（左或右）。发光镧系元素配合物具有独特的光物理性质，其左旋和右旋发光的比例对周围环境的温度非常敏感。我们新开发的全固态CPL光谱仪是CPL光谱学的范式转变，由于其体积小和多功能性，它可以真正广泛应用。由于我们在仪器开发方面的专业知识和良好记录，将这种技术应用于高空间（光学精度）和时间（采集速度）分辨率的显微镜设置是很简单的。因此，我们建议构建和验证世界上第一个共聚焦激光扫描CPL显微镜（CPL-LSCM）。我们充分意识到使用传统光学显微镜进行深层组织成像是具有挑战性的。因此，我们还计划采用低能量、生物安全的近红外（NIR）多光子激活（MP）来增加组织的可观察深度。它将实现前所未有的活细胞对映选择性手性光学显微镜，并提供巨大的科学价值，为研究和跟踪内源性和工程生物探针的发射手性分子开辟新的视野。凭借我们在快速CPL光谱学方面的开创性工作，我们可以实现这一目标。我们的建议具有巨大的潜力，超越了与广泛和多样化的多学科科学界相关的好处。它还可能产生巨大的商业利益，开启现代活细胞光学显微镜的新篇章，并揭示以前未被探索的生化过程，这些过程从根本上支持生命和危及生命的疾病进展。我们的首要目标是将其建立为帮助回答基本问题的“研究工具”，例如，为什么生命建立在其手性氨基酸结构单元的一种特定（L）对映体上，而需要葡萄糖的相反（D）对映体。它还可以扩展我们对酶等复杂生物分子和细胞分裂等基本生物化学过程的理解，揭示手性在其中的明确作用和驱动力。

项目成果

Two-Photon Circularly Polarized Luminescence of Chiral Eu Complexes.

• DOI: 10.1021/jacs.3c05957
• 发表时间: 2023-11-22
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Willis, Oliver G.;Petri, Filippo;De Rosa, Davide F.;Mandoli, Alessandro;Pal, Robert;Zinna, Francesco;Di Bari, Lorenzo
• 通讯作者: Di Bari, Lorenzo

Hierarchical self-assembly in an RNA-based coordination polymer hydrogel.

• DOI: 10.1039/d3dt00634d
• 发表时间: 2023-05-02
• 期刊: Dalton transactions (Cambridge, England : 2003)

Distinguishing Molecular Mechanical Action from Photothermal and Photodynamic Behavior.

区分分子机械作用与光热和光动力行为。

• DOI: 10.1002/adma.202306669
• 发表时间: 2023
• 期刊: Advanced materials (Deerfield Beach, Fla.)
• 作者: Beckham JL

A Chirally Locked Bis-perylene Diimide Macrocycle: Consequences for Chiral Self-Assembly and Circularly Polarized Luminescence

• DOI: 10.1021/jacs.3c13191
• 发表时间: 2024-02-14
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Penty，Samuel E.;Orton，Georgia R. F.;Barendt，Timothy A.
• 通讯作者: Barendt，Timothy A.