Omics in 4D
TIMS adds 4th dimension of separation
Mass spectrometry (MS)-based proteomics has become a powerful technology for the identification and quantification of thousands of proteins. However, the coverage of complete proteomes is still very challenging due to the limited speed, sensitivity and resolution of current mass spectrometers.
被困的离子迁移谱法(蒂姆斯)是第一位and foremost a separation technique in gas phase, which resolves sample complexity with an added dimension of separation in addition to HPLC and mass spectrometry, increasing peak capacity and confidence in compound characterization. Equally as important, the TIMS device also serves to accumulate and concentrate ions of a given mass and mobility, enabling a unique increase in sensitivity and speed along with the additional dimension of separation.
Achieve near 100% duty cycle for shotgun proteomics
A near 100% duty cycle for high sensitivity, high speed shotgun proteomics can now be achieved with the dual TIMS technology. The novel design allows for ions to be accumulated in the front section, while ions in the rear section are sequentially released depending on their ion mobility. This process is called Parallel Accumulation Serial Fragmentation, or PASEF®.
The second generation dual TIMS analyzer is optimized for shotgun proteomics needs. Due to its unique geometry ions are released dependent on their mobility from the second section of the TIMS analyzer while the further incoming ions can be accumulated in parallel in the first part of the TIMS analyzer. With the parallel accumulation technology a duty cycle of nearly 100% is achieved resulting in nearly no ion loss.
考虑到速度，布鲁克专家重新设计了MS/MS技术，以满足shot弹枪蛋白质组学的要求。使用被困的离子迁移率光谱法分离肽离子，洗脱（〜100 ms）并在QTOF中检测到，生成TIMS MS热图。在Pasef中®method the same TIMS separation is used with the quadrupole isolating a certain ion species during its elution and immediately shifting to the next precursor. Parent and fragment spectra are aligned by mobility values. With the Parallel Accumulation Serial Fragmentation (PASEF®) Technology >100 Hz sequencing speed can be achieved. With the PASEF®method the MS/MS spectra quality of the low abundant peptides can be increased by selecting them several times.
PASEF®: the perfect fit for shotgun proteomics
由Pasef提供动力的timstof Pro®offers sequencing speed >100 Hz without losing sensitivity or resolution. This is achieved by synchronizing the quadrupole isolation mass window with the elution time of the specific peptide packages from the TIMS funnel.
Bruker has introduced the TIMS (Trapped Ion Mobility Spectrometry) technology in 2016 as a revolutionary ion mobility technology, achieving unprecedented levels of ion mobility resolution in an extremely compact device. The second generation of dual TIMS analyzer now enables PASEF®(Parallel Accumulation Serial Fragmentation) that is setting new standards of uncompromised speed, sensitivity and resolution for shotgun proteomics.
许多女士仪器used for shotgun proteomics require biweekly or monthly cleaning when run 24 hours a day on large sample cohorts, and performance degradation is noticeable over even shorter time periods. The superior robustness of the timsTOF Pro means that the instrument can be run 24/7 over many days or weeks without noticeable loss of sensitivity or other performance metrics.
The timsTOF Pro allows the analysis of hundreds of samples with minimal sample load (<200 ng) at sequencing speeds exceeding 100 Hz. All of this with uncompromised proteomic depth. This has changed the way proteomics is done, but it also unleashes a new constraint - data analysis. Modern researchers must process hundreds of samples of information dense content with the timsTOF Pro. For most pipelines this creates a bottleneck at the data analysis step. Bruker is innovating this step by introducing real-time database search capabilities called PaSER (Parallel database Search Engine in Real-time) which removes the data analysis hurdle. Using PaSER, as soon as a LC-MS run is completed, results are in hand or Run & Done.
实现高通量，高灵敏度的靶向蛋白质组学。PRM-PASEF®已经开发了采集方法来翻译并行积累序列碎片的优势（Pasef®) acquisition strategy to the targeted proteomics field.
The combination of integrated ion mobility with warp-speed MS/MS acquisition on the timsTOF Pro paves the way for deep, molecular level understanding of β-cell physiology and potential new targetable pathways for diabetes.
高通量4D-Proteomics™ – Application of dia-PASEF®and the Evosep One for short gradients
The timsTOF Pro offers a combination of two unique technologies, namely a 4th dimension provided by Trapped Ion Mobility Spectrometry (TIMS) to enhance ion separation and sensitivity and Parallel Accumulation Serial Fragmentation (PASEF®)