布魯克於ASMS2019上推出高度創新的新型質譜產品timsTOF fleX™質譜儀，包括適配於ESI timsTOF Pro™質譜的軟體可切換的MALDI源。這種新型ESI/MALDI組合功能，可在單臺質譜上實現空間定位組學SpatialOMx™。timsTOF fleX配備布魯克獨有的10 kHz SmartBeam™ 3D雷射源，具有真正的像素保真度，可在高空間解析度下進行快速、無標記的MALDI成像，同時完全保持timsTOF Pro在ESI模式下無與倫比的4D蛋白質組學和表型組學的靈敏度。
利用這種獨特的SpatialOMx方法，研究人員可以深入洞悉MALDI成像組織中的空間分子分布，從而指導4D組學分子表達研究，例如蛋白質、低水平癌抗原肽、脂質、聚糖、代謝物或那些用傳統染色或標記技術無法觀察到的分子。在MALDI指導下的SpatialOMx方法中，使用者可用隨後由ESI-TIMS/PASEF的dda（數據依賴採集），或者dia（非數據依賴採集）4D蛋白質組學或4D脂質組學/代謝組學的數據，來開展細胞亞群的特異性靶向研究。兩者現在都可以在同一臺強大的儀器上進行，即timsTOF fleX，具有推進單細胞生物學研究所需的超高靈敏度，成為用RNA測序（RNA-seq）進行單細胞轉錄組學研究的完美補充。
The microenvironment of cancers contains a combination of healthy cells, tumor cells, connective tissue, blood vessels, and inflammation in different ratios at different time points. Each of these components will have their own unique-molecular signature of compounds. Researchers who study disease states rely heavily on interpreting tissue pathology and creating these maps within the context of biomolecules bridges the gap between traditional OMICS and understanding disease.
Cancer cells and other disease states have significant genetic and epigenetic modifications that influence the genomic expression cascade. Whether you’re looking at the proteome, lipidome or metabolome, the spatial distribution of compounds contains valuable Information for understanding your sample. If certain compounds are highly spatially concentrated or if molecules are co-distributed on specific structures, this vital information is missing when only examining homogenized samples. OMICS based biomarker discovery can be limited without the benefit of spatial information to add important contextual clues as tissue-level communications networks are integral to cancer growth.
The timsTOF fleX is a versatile instrument for probing the molecular content of your sample. Built on Bruker’s pioneering timsTOF Pro platform, the timsTOF fleX is a fully functional high speed, high sensitivity ESI instrument for all your X-Omics analyses, combined with a high spatial resolution MALDI source and stage specifically designed for resolving molecular distributions and bringing a spatial dimension to OMICS analyses. Transform proteomics analysis into spatial proteomics, lipidomics into spatial lipidomics, and metabolomics into spatial metabolomics.
Mapping complex molecular networks exposes intricate systems of interacting molecules within intracellular networks. Using SpatialOMx to create insightful atlases of communication mechanisms means learning more about the way cells communicate with each other to support change:
how cells interact with the local microenvironment to formulate promotion and mediation processes
how molecular networks of specific cells interact to perform specific biological functions
what effect epigenetic modifications have within these networks.
The timsTOF fleX – the flexibility to switch between spatial metabolomics, spatial lipidomics or spatial proteomics with ease.
Built on the standard for shotgun proteomics, the timsTOF fleX combines best in class 4D X-Omics with Bruker’s cutting edge MALDI Imaging technology, including SmartBeam 3D laser optics for fast measurement all in one platform. A dual source instrument ideal for SpatialOMx, timsTOF fleX conducts robust ESI measurements and spatially resolves a wide range of molecules directly from tissue using one platform. No single instrument has before provided access to both essential capabilities for the most advanced OMICS researchers.
SpatialOMx demands PASEF enabled LC-MS/MS for deep 4D OMICS combined with the ability to create molecular images of tissue sections with MALDI Imaging. To do this under one platform, Bruker engineers started with the award winning TIMS dual-stage ion funnel and incorporated target stage and laser elements from the rapifleX axial TOF instrument. During operation, software activation of the SmartBeam 3D laser is the only change in the source region. No complicated changeover means making zero compromises in productivity and the ability to move effortlessly from world class OMICS identification and quantification workflows to creating high definition molecular maps of tissue sections to see what matters most.
Since ions produced by MALDI and ESI travel the same path to the detector from the source, MALDI workflows can take advantage of the most advanced features found in the timsTOF Pro, including TIMS ion mobility separation based on the collisional cross section of detected molecules and high-speed high-sensitivity PASEF fragmentation. Tuning and calibration can be performed in ESI mode and used for the MALDI experiment for additional ease of optimization.
MALDI Imaging is a label-free tool for examining the distribution of molecules directly from thin tissue sections. Applicable for examining a wide range of analytes, including metabolites, lipids or glycans, and seamlessly compatible with microscopy workflows, MALDI Imaging can identify regionally specific compound distributions for SpatialOMx analyses. Use Bruker’s IntelliSlides for automated MALDI Imaging and SpatialOMx workflows on the timsTOF flex.
Besides the ability to generate high definition 「True Pixel」 MALDI images, the SmartBeam 3D laser source provides rock solid laser guided targeting down to 10 μM with Bruker’s new zoom mode technology. Users now have the flexibility to adjust their laser spot size and spacing to accommodate a wider range of granularity depending on biological features of interest.
Combining X-Omics performance with high MALDI sensitivity
Whether it’s proteomics, lipidomics, glycomics, or metabolomics, the timsTOF fleX is the ideal platform for deep SpatialOMx analyses. Use patented SmartBeam™ 3D technology to conduct fast, label-free MALDI Imaging for mapping the distribution of molecules in your sample and identify regions of interest for more specific analyses. Apply PASEF powered LC-MS/MS for the highest level of identifications and for greatest confidence in your results.
Combining the power of trapped ion mobility spectrometry (TIMS) and Parallel Accumulation Serial Fragmentation (PASEF), the timsTOF fleX can deliver quantitative information from smaller amounts of highly complex mixtures. Using less than 200 ng sample load and a 90 min gradient length, more than 5400 protein groups can be identified from a typical human cell line lysate. The high sensitivity reduces sample preparation costs and decreases time lost to maintenance.
With existing protocols for lipidomics, metabolomics or glycomics, MALDI Imaging spectra can be used as a molecular fingerprint for specific regions, or to investigate how compounds are localized across an entire sample for biomarker discovery. This can also be used for investigation of temporal changes, quantification, and correlation of compounds to tissue states. The timsTOF fleX allows for fast high mass resolution MALDI imaging – with the SmartBeam 3D and specialized MALDI source and stage, a high spatial resolution measurement of lipids from a rat brain section consisting of 370,000 pixels takes less than 5 hours.
Traditional workflows that sample small tissue sections (<100 μM) provide deep molecular details for a variety of chemical classes including RNA/DNA, proteins, glycans, lipids and metabolites. However, processing times are long and tissue excise locations are determined with optical images that lack molecular signal information. SpatialOMx on the timsTOF fleX rapidly provides a high definition, label-free overview that merges chemical information with vital morphological context. Choose your regions of interest manually or use SCiLS enabled automated segmentation to know exactly where your extraction will have the highest impact.
Using Bruker’s industry leading software, it is now possible to annotate target molecules directly from tissue. Simply load your data into SCiLS Lab, define regions of interest, and export the peak lists to MetaboScape. Annotate your peaks using databases or compound lists from LC-MS/MS analyses, then review and export your annotations back to SCiLS for visualization. From SCiLS you can then visualize results with pathways and familiar nomenclature rather than mass labels to reduce the time from data to results.
南卡羅來納醫科大學蛋白質組學中心主任Richard Drake教授表示：「使用新型timsTOF fleX系統運行的樣本，其數據在空間解析度和聚糖覆蓋深度方面是前所未有的。聚糖已成為組織和血清中潛在的臨床標誌物，用以監測身體的免疫狀態，正常或非正常的衰老，timsTOF fleX的獨特功能將大大加速該研究進程。 timsTOF fleX使得我們可將為癌症和免疫療法的組織和生物流體分析開發的方法匯集在一個平臺上。我可以看到這款儀器在許多研究領域中的無限應用，比如快速聚糖組織成像和生物流體4D組學分析。」
布魯克道爾頓執行副總裁Rohan Thakur博士補充說：「大多數組織蛋白質組學研究融合了來自不同亞群的細胞，其不期望的平均效應掩蓋了許多重要的生物學和病理生物學信息。在timsTOF fleX上，MALDI引導的SpatialOMx方法能夠在空間上對指定的組織區域進行分析，從而允許隨後的4D蛋白質組學選擇性地對細胞類型的亞群進行靶向分析。功能強大、具有超高靈敏度的timsTOF fleX，在同一儀器上還提供nanoLC-TIMS-MS/MS，從而彌合了分子組織成像和體液分析間的鴻溝。這種獨特的組合將使timsTOF fleX成為寶貴的研究工具，用於空間分辨的4D蛋白質組學和表型組學，並推動單細胞生物學、藥物蛋白質組學和大細胞數量或患者隊列的轉化4D交叉組學的工作流程。」
Helmholtz藥物研究所（HIPS）助理教授Daniel Krug博士評論說：「我們在藥物研究和空間代謝組學領域使用質譜技術；尤其是分析新型天然產物的粘細菌次級代謝組。我們實驗室多年來一直使用布魯克的儀器和軟體，特別是SCiLS Lab和MetaboScape；新分子成像工作流程集成了這兩種軟體解決方案，極大地簡化並加速了整個MALDI成像流程。該流程將離子圖像轉換為分子圖像，並提高了注釋分子式的置信度。」
創新點：
布魯克推出新型timsTOF fleX™質譜儀，包括適配於ESI timsTOF Pro™質譜的軟體可切換的MALDI源。這種新型ESI /MALDI組合功能，可在單臺質譜上實現空間定位組學SpatialOMx™。 timsTOF fleX配備布魯克獨有的10 kHz SmartBeam™ 3D雷射源，具有真正的像素保真度，可在高空間解析度下進行快速、無標記的MALDI成像，同時完全保持timsTOF Pro在ESI模式下無與倫比的4D蛋白質組學和表型組學的靈敏度。南卡羅來納醫科大學蛋白質組學中心主任Richard Drake教授表示：「使用新型timsTOF fleX系統運行的樣本，其數據在空間解析度和聚糖覆蓋深度方面是前所未有的。聚糖已成為組織和血清中潛在的臨床標誌物，用以監測身體的免疫狀態，正常或非正常的衰老，timsTOF fleX的獨特功能將大大加速該研究進程。 timsTOF fleX使得我們可將為癌症和免疫療法的組織和生物流體分析開發的方法匯集在一個平臺上。我可以看到這款儀器在許多研究領域中的無限應用，比如快速聚糖組織成像和生物流體4D組學分析。」