Metabolomics

Metabolomics

The Metabolomics Unit is one of the newest extensions of the CCP expanding the method portfolio of the Metabolic and Clinical Biochemistry Units. The analysis of blood is part of our standard first-line phenotyping. We analyze about 30 biochemical parameters following IMPC IMPReSS. More info

Measuring only a limited number of biochemical markers, increases the risk of missing the physiological impact of a studied gene or a treatment, or the early onset of a disease. Therefore, we implemented state-of-the-art metabolomics technology to analyze blood for molecules that may even give a hint to the mechanistic basis of a disease-relevant phenotype.

MALDI imaging, which is part Metabolomics Unit, offers the mass spectrometry technology applied to histological samples, as it enables measurements of multiple analytes in single experimental run in spatial context, correlated with histological annotations. Currently we are measuring metabolites, lipids and proteins – mainly in various histological samples (healthy tissue, tumours) but biological fluid as well (plasma, urine). The unit is equipped with state-of-the-art imaging mass spectrometer and matrix sprayer for fine-crystal sample covering which in tandem enables nearly cell-size resolution (5 µm). With connection to MALDI Biotyper ® database we can identify broad portfolio of bacteria at high speed and confidence.

Our instrument is of TOF/TOF type, so the analyte may undergo fragmentation for further species identity confirmation (LIFT technology). Imaging data are process with dedicated software for MALDI imaging datasets (SCiLS Lab).

Standard services Metabolomics

Discovery of diagnostic biomarkers, drug metabolization and their effects on whole metabolome, and progression of diseases are examples where studying metabolite profiles provided additional value also regarding translation to human disease.

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Standard Services MALDI Imaging Analysis

Sample Processing

With collaboration of our Histopathology Unit we are preparing histological cryo-sections from flash-frozen or paraffin-embedded tissues and placing them onto conductive histological size glass (ITO glass).

MALDI image of Mouse Embryo E18.5. Cryocuts coated with Norharman matrix, spectra acquired in negative mode. Lipids and metabolites are merged into single image.

Matrix Spraying

Next step in sample processing is spraying the matrix with TM-Sprayer machine in order to obtain homogenous crystal layer of matrix substance. This enables the extraction of analyte without loss of spatial resolution. Our sprayer is capable to spray virtually any matrix within broad range of conditions (matrix type and concentration, temperature, solvent composition, gas pressure) in appreciable speed.

MALDI Image of Mouse Ovary. Cryocuts were sprayed with Norharman matrix, spectra acquired in negative mode. Different lipids are displayed as single image and as merged image, respectively.
MALDI image of PDX. Tumour xenograft orthotopicaly implanted to colon. Sample sprayed with 9-aminoacridine matrix. Different lipids are displayed as a single and merged image, respectively.

MALDI Measurements

Rapiflex mass spectrometer is dedicated for MALDI imaging with high speed of image acquisition (laser frequency 10 kHz) with high spatial resolution (down to 5 µm). These enable to process even large samples or many tissue cuts to be analyzed in shortest time as possible and to utilize even matrices which quickly evaporates in high vacuum (such as DHB or Norharmane). Rapiflex mass spectrometer has good sensitivity and resolution. LIFT technology may be applied for fragmentation and confirmation of sample identity.

Data Processing

With high speed of data acquisition and high spatial resolution of images resutling datasets may become large. SCiLS Lab software together with dedicated processing PC station are fast enough to deal this task. SCiLS Lab software offers the possibility to display the mass spectrometry imaging data, correlate the images with histological staining and performs various statistical computations on multiple sections or datasets in one run.

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Custom services Custom Analysis

Pharmacokinetics of drugs after oral or intravenouse application

Metabolites profiling

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