MALDI Guided SpatialOMx on the timsTOF fleX

Match PASEF powered LC-MS/MS identification with label-free spatial localization to decode the molecular make up of your sample

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 10µm 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.

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. 

Specifically designed to simplify MALDI Imaging workflows, IntelliSlides are the optimal tool for mapping molecules across your samples.  Your effortless time-to-result has never been faster.

IntelliSlides are the ideal sample target for MALDI Imaging. Potential sources of inefficiency are removed by having permanent inscriptions on the conductive surface. These inscriptions streamline the User experience by aiding sample tracking and indicating where to place the tissue sample. Always correctly labelled, always oriented properly, IntelliSlides simplify the way to your results.

Perfect for SpatialOMx

In SpatialOMx workflows on the timsTOF fleX, MALDI Imaging is used to map molecule distributions across tissue sections, thereby identifying regions of interest and focusing X-Omics analyses to specific cell populations. With IntelliSlides and specialized software, the image registration process on the timsTOF fleX is automated at the touch of a button. Quickly aligning the target sample with the timsTOF fleX laser optics, the workflow set up is brought down to three minutes with IntelliSlides, allowing you to more rapidly and accurately match the chemical information to every pixel. 

IntelliSlides have the same dimensions of standard microscopy slides and are compatible with common histology stains and procedures, such as hematoxylin and eosin. With the permanent inscriptions, there’s no more concern that registration marks (also known as ‘teach marks’) will dissolve in histology solvents, allowing for easier co-registration of post-measurement stained tissues with the corresponding MALDI dataset.

Features and Benefits of IntelliSlides 

• Fulfills all requirements for reliable MALDI ionization
• All inscriptions on conductive slide surface
• Permanent registration marks (teach marks) indicate where to place the sample and are used for alignment within MALDI instrument
• Unique serial number and barcode to simplify sample tracking
• Automated image acquisition set up with fleXimaging 5.1 and above
• Compatible with histology and microscopy techniques
• IntelliSlides are a single-use consumable to avoid potential sources of contamination