FTMS Booster X1 High-performance data acquisition system for Orbitrap FTMS

Transcription

1 FTMS Booster X1 High-performance data acquisition system for Orbitrap FTMS

2 What is FTMS Booster? The Spectroswiss FTMS Booster X1 is a high-performance data acquisition and analysis system based on state-of-the-art FPGA (field-programmable gate array) technology. Together with our Peak by Peak software, FTMS Booster X1 is a complete state-of-the-art solution for acquisition of time-domain signals (transients), real-time digital signal processing, trigger decoding, and off-line data visualization and processing. Compatible with any Orbitrap FTMS platform (LTQ Orbitrap, Exactive, and Fusion series), FTMS Booster X1 gives your existing instrument a quantum boost in data quality and processing sophistication, helping you to take on the most complex and challenging applications. 2

3 Data Acquisition Module Embedded computer Processing and visualization of time-domain signals and mass spectra. High-bandwidth chassis PXI Express backplanes for high-speed data transfer. Low footprint and lightweight. High-performance digitizer / FPGA High sample rate analog-to-digital conversion, in-line digital signal processing and advanced triggering. Differential amplifier Low-noise, high-bandwidth amplification of transients. 3 Data storage buffer High-speed streaming and buffering of both transients and mass spectra.

4 New Generation Architecture Reduced Noise for Improved Sensitivity, Dynamic Range Full Transient Recording for Improved Resolution, Sensitivity Reduced Phase Distortions for Improved Resolution, Mass Accuracy High-throughput in-line FPGA technologies are the basis of the new architecture in FTMS Booster X1. Compared to traditional systems, the fidelity, accuracy and extent of the acquired time-domain data are substantially improved. This translates directly into improved analytical performance. Signal conditioning ADC FPGA High-performance Data bus Host PC Block diagram of FTMS Booster X1. The analog signal from the Orbitrap FTMS passes through our signal conditioning sub-system, before digitization in the analog-to-digital converter (ADC). The digital data stream is continuously processed by a high-performance FPGA chip with proprietary firmware. The high definition but compact digitized transients are transferred to the embedded (host) computer for processing and visualization. 4

5 Seamless Hardware and Software Integration High-Throughput Data Streaming Compatible with Any Orbitrap FTMS Open File Formats for Greater Flexibility Operates Parallel to Existing Data Acquisition FTMS Booster X1 is compatible with any contemporary Orbitrap FTMS instrument and requires no major modification or replacement of existing components. Both on-line and off-line software operate parallel to installed products. Two complementary data sets are simultaneously and synchronously acquired. Both Booster-produced and conventional transients or mass spectra can be processed by Peak by Peak and AutoVectis software tools. Booster data is synchronized with metadata produced by the existing software. Parallel, simultaneous acquisition and processing paths maximize instrument and analysis flexibility. 5

6 Improved Performance Owing to Full Transient Recording Reduced Phase Distortion On-line Absorption Mode FT The FTMS Booster X1 in combination with Peak by Peak software can increase the resolution, sensitivity, and limit of detection (LOD) in Orbitrap FTMS applications, including proteomics : Resolution performance of Orbitrap Fusion Lumos FTMS (Thermo Scientific) with and without FTMS Booster X1. Panel (A) shows total ion current (TIC) chromatogram and (B) single MS / MS scan from LC MS / MS experiment of HeLa tryptic digest. Panels (C) and (D) show expanded views into a selected MS / MS spectrum. The data are represented in (black) enhanced FT (eft, Thermo Scientific) and (red) absorption mode FT (aft, AutoVectis). Both mass spectra are normalized to the base peak of the eft mass spectrum. The threshold of nine standard deviations of noise (blue dashed line) was calculated from the aft mass spectrum. Sensitivity and LOD performance with Orbitrap Fusion Lumos FTMS (Thermo Scientific) with and without FTMS Booster X1. Panels (E) and (F) show expanded views. The data are represented in (black) enhanced FT (eft, Thermo Scientific) and (red) absorption mode FT (aft, AutoVectis). Both mass spectra are normalized to the base peak of the eft mass spectrum. Green arrows indicate appearance of additional analyte peaks in mass spectrum acquired with the FTMS Booster X1. The threshold of nine standard deviations of noise (blue dashed line) was calculated from an aft mass spectrum. 6

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8 Improved sensitivity, limit of detection and limit of quantitation owing to Full Transient Recording On-line Absorption Mode FT Full Profile Mode Mass Spectra Reduced Analog and Digital Noise The FTMS Booster X1 increases sensitivity, limit of detection (LOD), and limit of quantitation (LOQ) in comparison with standard built-in data acquisition systems for Orbitrap FTMS applications, including proteomics : A B C Sensitivity, LOD, LOQ performance with Orbitrap QExactive Plus FTMS (Thermo Scientific) with and without FTMS Booster X1. Panel (A) shows a total ion current (TIC) chromatogram, panels (B) and (C) each show mass spectra from LC MS / MS experiment of 6x5 reference peptide mix (Promega). The data are represented in (black) enhanced FT (eft, Thermo Scientific) and (red) absorption mode FT (aft, AutoVectis). Mass spectra are normalized to the base peak of the eft mass spectrum. Expected ratios of signal amplitudes of peaks in eft and aft mass spectra in panels (B) and (C) are four orders of magnitude. Green arrows indicate appearance of additional analyte peaks in aft mass spectra. Theoretical mass spectra of the two peptides from 6x5 LC MS / MS reference peptide mixture (Promega). Five isotopic clusters for each peptide represent four orders of concentration difference. Target peaks considered in the mass spectra above are shown in red. 8

9 Improved protein quantitation in proteomics FTMS Booster X1 and allied super-resolution signal processing (N-Plex Suite) and data analysis (Peak by Peak ) software increase productivity of Orbitrap FTMS in protein quantitation with isobaric mass tags, namely N-plex TMT and itraq technologies : FTMS Booster X1 acquires transients and absorption mode FT mass spectra in parallel to mass spectra acquisition with Orbitrap FTMS in enhanced FT mode (eft,.raw). Peak by Peak ensures synchronization between the two data sets. In case resolution provided by absorption mode FT is not sufficient, N-Plex Suite is used to perform super-resolution (least squares fitting, LSF) signal processing. Synergy of aft and LSF signal processing allows for 2-4 fold increase in resolution compared to eft, thus speeding up protein quantitation and allowing one to quantify more peptides / proteins per LC MS / MS experiment. Both methods, aft and LSF, demonstrate quantitatively-accurate performance even for very low SNR peaks. An example of 10-plex TMT protein quantitation with FTMS Booster X1 and on-line aft signal processing : 9

10 Improved sequence coverage and confidence in Top-Down Mass Spectrometry A workflow based on FTMS Booster X1 and Peak by Peak improves top-down protein analysis. Multiple consecutive LC MS / MS experiments are performed on a protein or a protein mixture of interest. Data are acquired in parallel using built-in electronics of Orbitrap FTMS and FTMS Booster X1 : mass spectra (.raw) and transients (.dat or.hdf5). Either spectral or transient summation is performed off-line using Peak by Peak software. Absorption mode FT signal processing of transients is enabled by AutoVectis software, co-developed with Dr. David Kilgour (Nottingham Trent University, UK). AutoVectis uses a genetic algorithm to optimize the phase correction function that relates ion frequency to the appropriate corrective phase angle to apply [1]. The final mass spectra or peak lists in the open formats (.csv,.mgf, or.mzxml) are provided for processing using external data analysis software. Top-down MS analysis of a ~20 kda hedgehog protein using electron transfer dissociation (ETD) on an Orbitrap Elite FTMS in 10 consecutive LC MS / MS experiments. A broadband ETD mass spectrum and its expanded views show the comparison of (black) 1 LC MS / MS experiment in eft representation and (green) transient-summed and aft-processed 10 LC MS / MS ones. The shown product ions were identified in the aft mass spectrum only. Dashed lines indicate corresponding noise thresholds (six standard deviations of noise). 1. Kilgour D. P. A., et al., Producing absorption mode FT-ICR mass spectra with non-quadratic phase correction functions. Rapid Commun. Mass Spectrom., 2015, 29,

11 Technical Data Content of delivery High-performance data acquisition system for commercial Orbitrap FTMS instruments System integration Data acquisition in parallel and synchronously with built-in commercial data acquisition systems Data file formats On-demand file formats of output data, including : HDF5, MGF, mzxml, and imzxml System installation System Trigger wire installation (1 x SMB connector): connection with the instrument s computer ; Signal wire installation (1 or 2 x SMB connectors): connection with the instrument s preamplifier Large data sets (e.g., mass spectra in full profile); High duty cycles (fast measurements, e.g. < 1 ms); Continuous recording of long transients (e.g. > 1 h) Embedded computer Intel Core i7 CPU 2.6 GHz, 8 GB RAM, USB 2.0 and 3.0, Ethernet OS : Windows 7 Professional Data visualization On-line and off-line representation of individual or summed transients, frequency or mass spectra Data storage buffer Analog bandwidth Differential input impedance Hardware inputs Dimensions Weight Power supply Warranty User-defined, e.g., 1-2 TB DC 100 MHz 1 MΩ, 50 Ω 3 x SMB connectors 185 x 300 x 210 mm 8.2 kg VAC, Hz, Max. 200 W 3 years 11

12 EPFL Innovation Park Building I 1015 Lausanne Switzerland T : info@spectroswiss.ch In a partnership with