Michael Steidel
@michaelsteidel.bsky.social
📤 268
📥 383
📝 83
#Cellzome
#TeamMassSpec
#Proteomics
opinions are my own
pinned post!
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/...
loading . . .
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.
https://pubs.acs.org/doi/10.1021/acs.jproteome.4c00932?articleRef=test
8 months ago
5
57
17
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?
about 9 hours ago
1
1
0
Astral Zoom hits >7,000 protein groups & 67,000 precursors — on a 500 SPD EvoSep ENO run.
www.biorxiv.org/content/10.1...
2 months ago
0
5
0
reposted by
Michael Steidel
Phil W
3 months ago
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...
loading . . .
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
https://github.com/pwilmart/quantitative_proteomics_comparison
3
41
14
Sciex in the game. Pretty impressed by ScanningSWATH data on the new ZenoTOF 8600.
#TeamMassSpec
3 months ago
1
11
2
#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...
loading . . .
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...
https://www.nature.com/articles/s41592-025-02719-x
3 months ago
1
3
0
reposted by
Michael Steidel
Nikolai Slavov
3 months ago
The videos from the 8th Single-Cell Proteomics Conference (#SCP2025) will be joining this growing YouTube playlist.
www.youtube.com/playlist?lis...
0
9
4
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?
4 months ago
2
0
0
www.biorxiv.org/content/10.1...
SPEC :The better SP3?
loading . . .
A Solid-Phase Extraction Capture (SPEC) workflow in nanoliter volumes for fast, robust and ultrasensitive proteomics
Despite great progress, sample preparation remains an area for improvement in proteomics, particularly for low-input samples where conventional protocols lead to losses and incomplete digestion. We pr...
https://www.biorxiv.org/content/10.1101/2025.05.31.657165v1
4 months ago
0
1
0
reposted by
Michael Steidel
Chris Ashwood
4 months ago
Introducing the new timsMetabo! A metabolomics focused timsTOF: - enhanced ion capacity of the dual-stage TIMS-MX ion funnel - Athen Ion Processor-equipped timsMetabo, up to 300 Hz PRM Also QSee software (a nightmare at talks, QC or Qsee?). As found by Biswapriya Misra
www.bruker.com/en/news-and-...
2
18
3
www.biorxiv.org/content/10.1...
timePlex enables time-domain sample multiplexing in LC-MS — boosting proteomics throughput up to 9× with no labels and minimal compromise in quant accuracy. Combine with plexDIA for 27 samples/run.
loading . . .
Increasing mass spectrometry throughput using time-encoded sample multiplexing
Liquid chromatography-mass spectrometry (LC-MS) can enable precise and accurate quantification of analytes at high-sensitivity, but the rate at which samples can be analyzed remains limiting. Throughp...
https://www.biorxiv.org/content/10.1101/2025.05.22.655515v1
4 months ago
1
4
0
reposted by
Michael Steidel
Ludwig Sinn
4 months ago
I am excited to see our performance assessment of the successor of Scanning SWATH on the Zeno TOF7600+ mass spectrometer - ZT Scan DIA - pre-printed (
www.biorxiv.org/content/10.1...
loading . . .
Performance Characteristics of Zeno Trap Scanning DIA for Sensitive and Quantitative Proteomics at High Throughput
Proteomic experiments, particularly those addressing dynamic proteome properties, time series, or genetic diversity, require the analysis of large sample numbers. Despite significant advancements in p...
https://www.biorxiv.org/content/10.1101/2025.05.06.652368v1
4
16
6
"when compared to DIA-NN, DIA-BERT demonstrated a 51% increase in protein identifications and 22% more peptide precursors"
www.nature.com/articles/s41...
loading . . .
DIA-BERT: pre-trained end-to-end transformer models for enhanced DIA proteomics data analysis - Nature Communications
Data-independent acquisition mass spectrometry (DIA-MS) has emerged as a key technology in quantitative proteomics. Here, the authors introduce DIA-BERT, a transformer model pre-trained on existing DI...
https://www.nature.com/articles/s41467-025-58866-4
5 months ago
1
4
0
#PipetocalypseNow
6 months ago
1
1
0
reposted by
Michael Steidel
PastelBio
6 months ago
Significant impact of consumable material and buffer composition for low-cell number proteomic sample preparation
chemrxiv.org/engage/...
---
#proteomics
#prot-preprint
0
5
2
Hey
#TeamMassSpec
#SingleCell
#Proteomics
, when using DDM in your workflow do you see it eluting from the column? If so at which m/z. Desperately looking for [M+H]+ at m/z 511.32
6 months ago
2
1
0
„Our method clusters peptides with similar quantitative behavior, providing a new approach to the protein grouping problem and enabling identification of regulated proteoforms directly from bottom-up data.“
www.biorxiv.org/content/10.1...
loading . . .
Tree-based quantification infers proteoform regulation in bottom-up proteomics data
Quantitative readout is essential in proteomics, yet current bioinformatics methods lack a framework to handle the inherent multi-level nature of the data (fragments, MS1 isotopes, charge states, modi...
https://www.biorxiv.org/content/10.1101/2025.03.06.641844v1
7 months ago
0
5
0
reposted by
Michael Steidel
Jan Broder Engler
7 months ago
New
#tidyplots
cheatsheet 🤩
tidyplots.org/cheatsheet
#rstats
#dataviz
#phd
0
39
10
Hi
#TeamMassSpec
, Does anyone have a clue how to assign multiple MS methods to single run via Bruker HyStar?
pubs.acs.org/doi/10.1021/...
loading . . .
Optimized Time-Segmented Acquisition Expands Peptide and Protein Identification in TIMS-TOF Pro Mass Spectrometry
We introduce here a novel approach, termed time-segmented acquisition (Seg), to enhance the identification of peptides and proteins in trapped ion mobility spectrometry (TIMS)-time-of-flight (TOF) mas...
https://pubs.acs.org/doi/10.1021/acs.jproteome.4c00690
7 months ago
2
0
1
reposted by
Michael Steidel
Fred Strathmann
7 months ago
Protein IDs for early
#PAMAF
performance on an Agilent QTOF (fyi - not our final detector)
#USHUPO
2
4
2
To those attending USHUPO: Is anyone willing to leak the absolute IDs in addition to the relative Mobilion IDs from the whitepaper?
#TeamMassSpec
#Proteomics
#PAMAF
7 months ago
1
3
0
reposted by
Michael Steidel
ProteomicsNews (Ben Orsburn)
7 months ago
The Trump administration is so on top of things that scientists EXTERNALLY FUNDED BY INDUSTRY WHO MAKE THE HHS MONEY are receiving termination letters. These fucking morons have no idea how the government even works. AT ALL
1
8
2
Popularity of current DIA data analysis tools (vendor-independent, based on citations of initial publication) Updated for 2024:
7 months ago
1
20
9
reposted by
Michael Steidel
Mariya Mardamshina
8 months ago
Calling all
#TeamMassSpec
experts. I’m planning to revisit the classic SCX fractionation protocol but ran into a challenge - I can’t find a vendor for syringes with metal springs to make stage tips. I’d be so grateful for any suggestions or advice you might have 🙏 *attaching picture as an example
4
3
7
reposted by
Michael Steidel
Vadim Demichev
8 months ago
DIA-NN 2.0 is released! We consider it the biggest step forward in the history of DIA-NN. On modern LC-MS almost all identifications are now peptidoform-confident, with major improvements e.g. for phospho. Some other cool things too:
github.com/vdemichev/Di...
loading . . .
Release DIA-NN 2.0 · vdemichev/DiaNN
We are excited to announce DIA-NN 2.0, the most significant milestone in the history of DIA-NN development. Key Breakthroughs Proteoform Confidence mode: DIA-NN 2.0 solves the long-standing chall...
https://github.com/vdemichev/DiaNN/releases/tag/2.0
6
150
41
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/...
loading . . .
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.
https://pubs.acs.org/doi/10.1021/acs.jproteome.4c00932?articleRef=test
8 months ago
5
57
17
reposted by
Michael Steidel
Samuel Pazicky
8 months ago
I am planning to switch from DDA to
#DIA
for our proteomics experiment on Orbitrap Exploris. Question: FAIMS yes or no? If I do not use FAIMS for library generation, can I use it for DIA measurements? Or the other way around?
#massspectrometry
#proteomics
#teammassspec
4
3
2
reposted by
Michael Steidel
Patricia Skowronek
8 months ago
🚀 Robust and high sensitivity
#proteomics
: Our Nature protocol demystifies
#PASEF
workflows and provides ready-to-use dia-PASEF & synchro-PASEF methods. Find out how to achieve 7,000 protein groups or 29,000 phosphosites in 21min. Let's explore!
#TeamMassSpec
#Bruker
doi.org/10.1038/s415...
1/đź§µ
loading . . .
https://doi.org/10.1038/s41596-024-01104-w1/đź§µ
4
35
11
Hi
#TeamMassSpec
, Is it expected that diaPASEF file size triples from Pro2 to Ultra2? (From 2 GB to 6 GB, 60 SPD) ...
8 months ago
1
0
0
reposted by
Michael Steidel
Witold Szymański
8 months ago
- Bruker did a full calibration remotely (didn't help) - I connected the CaptiveSpray Ultra to the other TimsTof Pro with the other nanoElute and the same effect so it is not the MS and not LC The problem is most probably in ITI or the whole CaptiveSpray source.
2
3
2
reposted by
Michael Steidel
Kyle Swovick
9 months ago
This may be an incredibly naive mass spec question, but what's a PSM for DIA data, particularly in Spectronaut? Is it simply referring to a confidently ID'd precursor?
3
2
2
Trying to estimate popularity of search engines. (Something widespread I missed out?) Based on citations/year of initial publication it seems as MSFragger would take over soon...
over 1 year ago
1
0
0
Hey
#TeamMassSpec
, Anyone there having both timsTOF SCP/Ultra +
#OrbiTOF
Astral in their lab? Would be interested in comparisons. Where do both have their respective sweet spots?
almost 2 years ago
1
0
0
Anyone having a good explanation why all Orbi systems require frequent cleaning while timsTOFs just continue to run stable ?
almost 2 years ago
3
1
0
Has anyone tried already cleaning-up proteins from Trizol samples using SP2/SP3/SP4 ? Acetone precipitation does not work robustly..
almost 2 years ago
1
0
1
Hey # TeamMassSpec, Have you ever considered DIA for small pulldown studies? I know that should definitely be one of the TMT sweetspots, but have not seen any back-to-back comparison so far..
almost 2 years ago
2
1
1
Hi
#TeamMassSpec
,
#timsTOF-users
Has anyone so far attempted to automatically trigger a first pass DIA-NN analysis of diaPASEF files directly from the HyStar queue?
almost 2 years ago
1
0
0
Hi
#TeamMassSpec
, Does anyone have a simple script at hand to filter fasta files for a list genes or uniprot accessions?
almost 2 years ago
0
0
0
reposted by
Michael Steidel
Lindsay K Pino
almost 2 years ago
FDR, as currently implemented, is probably broken. Short gradients, DIA, chimeric spectra, PTMs, emphasis on quant. Everyone talks about "how" to do FDR "right" but maybe we need to ask WHY we're doing it and if we should rethink the entire process based on the purpose.
3
12
3
you reached the end!!
feeds!
log in