Michael Steidel
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michaelsteidel.bsky.social
Michael Steidel
@michaelsteidel.bsky.social
#Cellzome #TeamMassSpec #Proteomics opinions are my own
Pinned

Weather’s going wild, and now your mass spec data’s a mess too?

Coindidence? Nope!

We reveal how weather-driven air pressure fluctuations impact diaPASEF-based high troughput proteomics - and how to fix it!

Check out our new paper! #diaPASEF #Weatheromics

pubs.acs.org/doi/10.1021/...
Impact of Local Air Pressure on Ion Mobilities and Data Consistency in diaPASEF-Based High Throughput Proteomics
Data-independent acquisition (DIA) on ion mobility mass spectrometers enables deep proteome coverage and high data completeness in large-scale proteomics studies. For advanced acquisition schemes such as parallel accumulation serial fragmentation-based DIA (diaPASEF) stability of ion mobility (1/K0) over time is crucial for consistent data quality. We found that minor changes in environmental air pressure systematically affect the vacuum pressure in the TIMS analyzer, causing ion mobility shifts. By comparing experimental ion mobilities with historical weather data, we attributed observed drifts to fluctuations in the ground air pressure. Moderate air pressure changes of e.g. fifteen mbar induce ion mobility shifts of 0.025 Vs/cm2. These drifts negatively impact peptide quantification across consecutively acquired samples due to drift-dependent abundance changes and increased missing values for ions located at the boundaries of diaPASEF isolation windows, which cannot be corrected by postprocessing. To address this, we applied an in-batch mobility autocalibration feature on a run-wise basis, leading to full elimination of ion mobility drifts.
pubs.acs.org
Reposted by Michael Steidel
We have released DIA-NN 2.7. Performance improvements (more proteins) and GUI improvements. In particular, added a data completeness filter for pathway analysis. Further feedback on how to improve statistics and visualisation in DIA-NN is very welcome!
September 16, 2026 at 10:26 AM
In the interest of a fair comparison, we intentionally made it unfair.

Orbitrap-TMT vs. timsTOF-diaPASEF across Kinobeads and 2D-TPP.
analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/...
Cross‐Platform Comparison of Chemoproteomics Workflows: Orbitrap‐TMT Versus timsTOF‐diaPASEF
Chemoproteomics aims to achieve precise and comprehensive quantification of protein–small molecule interactions, yet methodological comparisons across quantitative proteomics workflows remain scarce.....
analyticalsciencejournals.onlinelibrary.wiley.com
September 16, 2026 at 7:42 AM
Hi #TeamMassSpec,
Anyone out there already using EvoSep LUPO? First impressions?
July 29, 2026 at 7:33 AM
Reposted by Michael Steidel
We preprinted a method of measuring phosphosite stoichimetries (occupancies) in just two mass spectrometry acquisitions, using internal stable isotope labelled controls.
June 2, 2026 at 2:18 PM
#TeamMassSpec: any independent data on dual-column LC creating subtle batch effects in LFQ proteomics?

Thinking column A/B intensity shifts, missingness, RT drift, or condition-column confounding. Vendor claims are nice; real-world datasets would be nicer.
May 26, 2026 at 12:49 PM
#TeamMassSpec
Has anyone actually done a clean head-to-head comparison of TMT quant on AstralZoom-Orbi vs TOF?
May 21, 2026 at 5:03 PM
Reposted by Michael Steidel
Single-molecule peptide sequencing through reverse translation of peptides into DNA www.nature.com/artic...

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#proteomics #prot-paper
March 19, 2026 at 7:40 AM
Reposted by Michael Steidel
A Ground-truth validation of FDR & false localisation control in proteomics – a must-read from Stefan Tenzer’s lab!

The benchmarks are quite enlightening. We are pleased to see our DIA-NN 2.0 excelling in sensitivity - often by a wide margin - while controlling FDR and false localisation rates.
March 12, 2026 at 2:53 PM
Reposted by Michael Steidel
Solving the computational challenge of phosphoproteomics with 𝐏𝐡𝐨-𝐓𝐢𝐩: One-Pot Dephosphorylation for Rapid and Sensitive Analysis of DIA Phosphoproteomics Data. Now out in Analytical Chemistry!

Makes predicted phosphopeptide libraries 10x-20x smaller. Link below.
February 23, 2026 at 12:38 PM
Reposted by Michael Steidel
It's called the Agilent 6495D.
January 3, 2026 at 10:32 AM
Proteomic Ruler question:
In Wiśniewski et al., MCP 2014, the histone→DNA proxy seems implicit.
Is there any explicit reference stating that the Ruler uses only core histones (H2A/H2B/H3/H4) and excludes H1?
#proteomics #massspec
December 14, 2025 at 1:41 PM
Reposted by Michael Steidel
Bonus, info about DIA multiplex tags, up to 30-plex:
"trademark DXT for our DIA multiplex tags...advances have been made in DXT multiplexing since ASMS with the number of tags increased from 6 to 11 and with the potential to
increase these to beyond 30"
November 18, 2025 at 10:09 AM
Hey #TeamMassSpec,

Many non-human proteomics studies still search against taxon-filtered FASTAs.

❌ Redundant sequences
❌ Inflated search space
✅ Reference proteomes cut redundancy, improve annotation, and make results comparable.

👉 Time to move beyond taxon filters. #proteomics #massspec #uniprot
September 30, 2025 at 2:21 PM
Reposted by Michael Steidel
Without #2, a lower ion count is needed just to be sure that the full MS range is scanned, but with more accurate ion counts, you can go to the max S/N without losing ions on the edges.
This could also work for the Orbitrap Astral.

Bonus: DIAPASEF on Thermo - patentscope.wipo.int/search/en/de...
September 28, 2025 at 1:25 PM
Reposted by Michael Steidel
With 𝗗𝗜𝗔-𝗡𝗡 𝟮.𝟯.𝟬 Preview (Academia-only for now), we showcase the transformative new capabilities that have been developed in the past months. Download: github.com/vdemichev/Di...
September 26, 2025 at 9:52 AM
Hey #TeamMassSpec,

When you run proteomics on non-human species (mouse, rat, macaque, etc.) — which protein FASTA do you prefer?

Taxonomy-filtered UniProt (all entries)

Reference proteome (SwissProt+TrEMBL)

Ensembl/GENCODE

Something else?
September 25, 2025 at 12:41 PM
Astral Zoom hits >7,000 protein groups & 67,000 precursors — on a 500 SPD EvoSep ENO run.

www.biorxiv.org/content/10.1...
July 19, 2025 at 9:01 AM
Reposted by Michael Steidel
DIA, DOA, DUI, DDA, etc. Here is a comparisons of some quantitative proteomics methods from a POV you might not have seen before:
github.com/pwilmart/qua...
GitHub - pwilmart/quantitative_proteomics_comparison: Comparison of DIA to spectral counting and TMT quantitative techniques using animal lens studies
Comparison of DIA to spectral counting and TMT quantitative techniques using animal lens studies - pwilmart/quantitative_proteomics_comparison
github.com
July 4, 2025 at 5:53 PM
Sciex in the game.

Pretty impressed by ScanningSWATH data on the new ZenoTOF 8600.

#TeamMassSpec
June 27, 2025 at 10:13 AM
#TeamMassSpec, Any opinions on why not generally adding the relatively small yeast proteome to the anyway large human search space (*.fasta) as an internal FDR quality control?

www.nature.com/articles/s41...
Assessment of false discovery rate control in tandem mass spectrometry analysis using entrapment - Nature Methods
A theoretical foundation for entrapment methods is presented, along with a method that enables more accurate evaluation of false discovery rate (FDR) control in proteomics mass spectrometry analysis p...
www.nature.com
June 23, 2025 at 8:38 PM
Reposted by Michael Steidel
The videos from the 8th Single-Cell Proteomics Conference (#SCP2025) will be joining this growing YouTube playlist.

www.youtube.com/playlist?lis...
June 22, 2025 at 11:14 AM
Hi #TeamMassSpec #EvoSep
Anyone using WhisperZoom for standard inputs (~500 ng)?
Getting great data on timsTOF Pro Ultra (with ICC2), but repeatedly hit overpressure on Aurora columns - forcing me to discard them.
Anyone else seeing this?
June 7, 2025 at 6:56 AM