MtoZ Biolabs offers PCT-DIA Service that combines pressure cycling technology (PCT) with data independent acquisition (DIA) based quantitative proteomics to unlock deep proteome coverage from challenging and precious samples. The PCT-DIA Service is tailored for FFPE tissues, small biopsies, and limited clinical specimens where conventional lysis and DDA workflows often fail to deliver robust, reproducible data.

What Is PCT-DIA?

Pressure cycling technology (PCT) is a sample preparation strategy that uses cycles of very high hydrostatic pressure and ambient pressure to efficiently disrupt cells and tissues. Repeated pressure cycling helps break tough membranes, loosen crosslinked structures, and release proteins and peptides from samples that are difficult to lyse by conventional methods. This approach is especially useful for fibrous tissues, heavily crosslinked matrices, and formalin fixed paraffin embedded specimens.

Data independent acquisition, or DIA, is a mass spectrometry acquisition mode that systematically fragments all ions within predefined mass windows. Instead of selecting only a few precursor ions for fragmentation as in data dependent acquisition, DIA repeatedly scans the full mass range in segments and collects fragment ion information for all detectable species within each window. The result is a highly consistent map of fragment ion traces that can be used for peptide identification and quantitative comparison across many samples.

PCT-DIA combines these two technologies into a single workflow. PCT improves protein extraction and digestion efficiency from challenging or low input samples. DIA then captures fragment ion data in a comprehensive and reproducible manner, enabling quantitative proteomics even when sample amount is limited or tissue quality is suboptimal. Together, they enable proteome analysis for sample types that are difficult to handle with standard lysis and DDA based methods.

PCT-DIA Service at MtoZ Biolabs

MtoZ Biolabs has established a dedicated PCT-DIA workflow to support projects that rely on scarce, heterogeneous, or archived samples. You only need to share your project objectives and send us the samples. MtoZ Biolabs handles the rest, from PCT based protein extraction through proteolysis, peptide cleanup, DIA acquisition, data processing, and bioinformatics interpretation, delivering ready to use results.

Guo, T. et al. iScience. 2019.

Figure 1. PCT-DIA Proteomics

Why Choose MtoZ Biolabs

• Specialized PCT experience for FFPE, micro-biopsies, and other difficult samples, maximizing protein recovery from limited material.
• DIA based quantitative workflows that deliver deep coverage, high data completeness, and robust reproducibility across cohorts.
• Optimized end-to-end process from PCT extraction to LC-MS/MS, with strict QC at each step to ensure stable performance.
• Flexible study design tailored to clinical, translational, or basic research needs.
• Professional proteomics and bioinformatics team providing clear reports, figures, and biologically meaningful interpretation.

Applications of PCT-DIA Service

PCT-DIA at MtoZ Biolabs can be applied in many biological and clinical contexts, particularly where sample amount or quality is limiting. Typical applications include:

1. Biomarker Discovery and Verification in FFPE Cohorts

Mining archived tissue collections for disease associated protein signatures.

2. Mechanism of Disease Studies

Understanding pathway changes in oncology, neurology, inflammation, fibrosis, and other disease areas using small or heterogeneous samples.

3. Drug Target and Pharmacodynamic Research

Assessing target engagement and proteome level responses in preclinical or translational models where tissue availability is limited.

4. Retrospective Studies Using Archived Specimens

Leveraging stored FFPE samples from clinical trials or registries for new proteomic analyses.

5. Spatially Informed or Microdissected Sampling

Analyzing microdissected tumor regions, border zones, or specific anatomical compartments where only tiny samples can be collected.

FAQ

Q1: What types of samples are suitable?

PCT-DIA is especially suited for difficult or low-input materials, including:

• Fresh tissue: ≥ 1-2 mg per sample

• Biopsy tissue: ≥ 1-2 mg per sample

• Pollen or fish fry: ≥ 1 mg per sample

• Cells: ≥ 5×10⁵ cells per sample

• FFPE: about 9 slices per sample (10 μm, total ≥ 1 mg)

If your available material is slightly below these amounts or of another type, please contact us to evaluate feasibility.

Q2: How should I prepare my samples?

1. FFPE tissues

• Provide intact blocks or clearly labeled sections on slides.

• Indicate tissue type, diagnosis, and region of interest if available.

2. Fresh frozen tissues and biopsies

• Snap freeze immediately after collection.

• Store at -80 °C and avoid repeated freeze thaw cycles.

3. Cell pellets

• Wash to remove excess media if possible.

• Spin down, remove supernatant, snap freeze, and store at -80 °C.

4. General recommendations:

• Use low binding tubes and avoid strong detergents, high salt, or stabilizers unless discussed in advance.

• Ship all samples on dry ice in insulated containers.

• Include a detailed sample list with IDs, groups, treatments, and basic clinical or experimental information.

Detailed requirements and recommendations can be found in the Sample Submission Guidelines for Proteomics.

Q3: What is the service general workflow?

Q4: What data formats are provided?

• Raw LC-MS/MS files in instrument specific formats
• Peptide and protein identification and quantification tables in Excel or CSV
• Differential analysis and pathway or functional enrichment results in spreadsheet format
• Publication ready figures such as heatmaps, volcano plots, and PCA plots in high resolution PNG or TIFF
• A structured PDF report summarizing workflow, QC metrics, key quantitative results, and main biological interpretations
• Additional formats and customized deliverables can be provided on request.

Start Your Project with MtoZ Biolabs

PCT-DIA Service from MtoZ Biolabs is designed for situations where traditional sample preparation and acquisition strategies are not enough, especially for FFPE tissue, scarce biopsies, and other difficult materials. If you are planning biomarker discovery, mechanism studies, or translational research with limited or archived samples, our team can help you unlock their proteomic potential.

Contact us to discuss your project goals, receive a tailored proposal, and turn challenging specimens into high quality, quantitative proteomics data.