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Improve DIA performance on Orbitrap mass spectrometers

Improve DIA performance on Orbitrap mass spectrometers

Improve DIA performance on Orbitrap mass spectrometers

Access the Velocity DIA information kit, infographic, and other tools to enhance DIA performance on Orbitrap mass spectrometers.

Access important DIA tools ›

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Faster throughput with greater proteome coverage for LFQ

The Thermo Scientific Orbitrap Astral Mass Spectrometer combines the  proven performance of Orbitrap technology with the speed (200 Hz) and sensitivity of the novel Astral mass analyzer, enabling faster throughput and deeper proteome coverage in label-free quantitation (LFQ) experiments . The mass spectrometer enables analysis of thousands of samples without loss in performance, delivers accurate and reproducible quantitation of thousands of proteins. 

Faster throughput with greater proteome coverage for LFQ

The Orbitrap Astral mass spectrometer combines the  proven performance of Orbitrap technology with the speed (200 Hz) and sensitivity of the novel Astral mass analyzer, enabling faster throughput and deeper proteome coverage in label-free quantitation (LFQ) experiments . The mass spectrometer enables analysis of thousands of samples without loss in performance, delivers accurate and reproducible quantitation of thousands of proteins. 

Label-free relative quantitation instruments
 Orbitrap Ascend
Orbitrap Ascend Tribrid Mass Spectrometer

Orbitrap Exploris 480 Mass Spectrometer

Orbitrap Exploris 240 Mass Spectrometer
Orbitrap Eclipse
Orbitrap Astral Mass Spectrometer
AdvantagesObtain maximum insights on complex samples. Facilitate advanced proteomics workflows: label-free quantitation, whole proteome profiling and single-cell proteomics with faster data acquisition. Orbitrap Ascend Mass Spectrometer’s new innovations deliver ultimate flexibility in experimental scope, accelerating your path to new, impactful results, to drive your science beyond today’s discovery.Get unparalleled insights from untargeted proteome profiles to targeted proteomics experiments with industry-leading single-cell sensitivity and accuracy. Curated workflows accelerate large-scale studies, delivering high-quality data and time savings for actionable outcomes.Optimized proteomics solutions for small-to-large-scale studies with fast turnaround times and best-in-class performance in a compact footprint. From protein identification to multiplexing, expand your capabilities with everyday versatility.By combining the proven performance of the Orbitrap mass analyzer with the speed (200 Hz) and sensitivity of the novel Astral mass analyzer, the Orbitrap Astral mass spectrometer enables faster throughput and deeper proteome coverage.
Resolving PowerUp to 480,000 (FWHM) at m/z 200Up to 480,000 at m/z 195Up to 240,000 FWHM at m/z 200Orbitrap resolution: settings range from 7500 to 480,000 (FWHM) at m/z 200

Astral resolution: 80,000 (FWHM) at m/z 524
Scan SpeedOrbitrap mass analyzer MSⁿ up to 45 Hz
Ion trap MSⁿ up to 50 Hz
Up to 40HzUp to 40HzOrbitrap MSn acquisition rates up to 40 Hz

Astral MS2 acquisition rates up to 200 Hz
Mass RangeStandard mass range m/z 40–2000, mid-mass range m/z 200–6000, and optional HMRn+ mass range m/z 500–16,00040 to 6,000 m/z (up to 8,000 m/z with the BioPharma option)40 to 6,000 m/z (up to 8,000 m/z with the BioPharma option)Orbitrap: Standard mass range m/z 40–6000, m/z 40–8000 with BioPharma option

Astral: Standard mass range m/z 40–6000
Dynamic Range>5000 with a single OTMS spectrum>5000:1>5000:1Orbitrap: >5000 within a single Orbitrap mass spectrum

Astral: >1000 within a single Astral mass spectrum
Mass AccuracyInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMSOrbitrap: Internal <1 ppm RMS; External: <3 ppm RMS

Astral: External <3 ppm RMS

Obtain broad peptide coverage

During label-free relative quantitation workflows, digested peptides are separated by LC prior to MS. RP separation on Thermo Scientific low-flow systems integrates well with label-free quantitation workflows and combines seamlessly with Thermo Scientific Orbitrap mass spectrometers. The Thermo Scientific Vanquish Neo UHPLC System is the system of choice. The Vanquish Neo UHPLC System combines an unrivaled degree of innovation to deliver 24/7 reproducible separations of complex mixtures at maximum performance for a variety of high-sensitivity LC-MS workflows



Software for label-free quantitation

Discover novel protein biomarkers

Thermo Scientific Proteome Discoverer Software, with its new label-free quantitation node, provides an easy-to-use automated solution for evaluating large amounts of LC-MS data generated by label-free experiments. The label-free quantitation node also features a full suite of quantitative capabilities previously only available for isotopically labeled workflows. The combination of label-free quantification workflows with the scaling, normalization, and study management features of Proteome Discoverer software offers powerful means for analyzing highly complex data.


Label-free quantitation workflow


Two different LFQ approaches to identification data acquisition

LFQ Data Dependent Analysis (DDA)

In a LFQ DDA approach, the ions for a given m/z range are individually isolated and fragmented. The quantitation involves extracting peptide chromatograms (MS 1 precursor ion) from LC-MS runs and integrating peak areas over the chromatographic time scale or using the intensity at the highest point of the chromatographic peak. The resulting areas or intensities are compared across sample set (i.e. control vs experimental) for quantitation.

In traditional data-dependent workflows, consistent precursor and thereby protein quantitation can be challenging to achieve due to the stochastic sampling nature of the mass spectrometer. Innovations in software algorithms now extract LC-MS peaks in the raw data files and map them to identified spectra, detecting these features across LC-MS data files using retention-time alignment and feature linking, minimizing "missing data points" and maximizing quantitative insights.

LFQ Data Independent Analysis (DIA)

In a LFQ DIA approach, a precursor mass range is selected and then divided into a series of relatively wide isolation windows: for example, 25 m/z each. Chimeric MS/MS data is acquired from all detected precursor ions in the first isolation window and then repeated for each consecutive, adjacent isolation window until the entire precursor mass range is covered. MS/MS spectral libraries are used to identify peptides of interest from the acquired data.

DIA significantly increases coverage and reproducibility by acquiring MS/MS data from all detected precursor ions; it also makes retrospective data analysis possible. LFQ DIA involves extracting peptide chromatograms (MS 1 precursor ion or MS 2 fragment ions) from LC-MS runs and integrating peak areas over the chromatographic time scale, and resulting areas are compared across the sample set for quantitation. The samples that are part of the comparison study are run individually.

Thermo Scientific Velocity LFQ DIA Platform

Velocity LFQ DIA workflow on an Orbitrap Exploris 480 mass spectrometer

Learn about the qualitative and quantitative performance of the Velocity LFQ DIA workflow on an Orbitrap Exploris 480 MS for large-scale proteomics analysis.

 Download technical note ›

Velocity LFQ DIA workflow for Orbitrap Ascend Tribrid MS

Velocity LFQ DIA workflow on an Orbitrap Ascend Tribrid mass spectrometer

Learn about the qualitative and quantitative performance of the Velocity LFQ DIA workflow on an Orbitrap Ascend Tribrid MS for large-scale proteomics analysis.

 Download technical note ›


Thermo Scientific Velocity LFQ DIA Platform

Explore a new dimension in proteomics quantitation

The new Velocity LFQ DIA platform sets a new standard for quantitative accuracy, precision and data completeness at deep proteome coverage for confident characterization..

Learn more ›