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.
TIMSTOF PRO使用平行的积累序列碎片(Pasef®)采集方法,以使用低样本量,提供极高的速度和灵敏度,以达到shot弹枪蛋白质组学和磷蛋白组学的新深度。
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.
在Pro中,高级分段四极质量滤波器用于高离子传输和隔离效率。四极质量位置与TIMS分析仪的特定离子的洗脱时间同步。由于其超高的质量位置切换时间(<1 ms),因此可以为PASEF提供最佳性能®方法。
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.
唯一的pasef®- 可增强的质谱仪
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.
卓越的鲁棒性,无需清洁
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.
Maxquant/Perseus和Peaks Studio数据处理
开放文件格式使研究人员可以直接与原始数据合作,并使用行业领先的软件。Maxquant已适应用于管理空间中的4维(4D)特征,该空间由保留时间,离子移动性,
质量和信号强度有益于肽,蛋白质和翻译后修饰的识别和定量。Peaks Studio将从头测序与传统数据库搜索结合在一起,并经过优化用于处理TimStof原始数据。
PRM-PASEF®:启用高通量,高灵敏度针对性蛋白质组学
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.
原代胰岛素分泌胰腺β细胞的蛋白质组学询问
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®)
提高离子利用率效率和数据采集速度。
“我们现在知道,在二维分析时(保留时间和m/z)时,肽混合物仍然非常复杂。在原则上,添加一个维度应该为我们带来很长的前进。除了分离的附加维度,timstof Pro为我们提供了极高的速度和灵敏度,以更深入蛋白质组并使用较少的样品材料。”
仅用于研究。不用于临床诊断程序。