SAMPLES

  • Blood and Urine

    Blood and Urine
    50 – 500 µL

  • Animal tissues

    Animal tissues
    20 – 100 mg

  • Culture cells

    Culture cells
    106 -107 cells

  • Microorganism

    Microorganism
    108 -109 cells

  • Others

    Others
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Because the metabolome includes a variety of compounds with different chemical properties, sampling procedures with stability and reproducibility is key to obtaining reliable data.

We can provide established procedures for a variety of sample types, and our study coordinator can assist your sample preparation depending on the study design.

Sample type Example Required volume*2 Preparation*3
Blood, Body fluids Plasma, Serum, Urine, CSF, fecal extract etc. 50 ~ 500 µL Frozen and shipped
Animal tissue Liver, Brain, Adipocytes, Muscle, Nerve, vessel etc. 30 ~ 100 mg
Plant tissue Leaf, Fruit, Loot, Alga
Culture cells*1 Culture / Primary cell line,
Hemocyte, Sorted cells
2 ~ 5 x 106 cells Extracted in your Lab
(or frozen and shipped)
Microorganism*1 Bacteria, Yeast, Fungus,
Microbiome etc.
1 ~ 10 x 108 cells
5 ~ 20 OD600 x mL
Medium etc. Culture medium, Beverage etc. 50 ~ 100 mL Frozen and shipped
Others Food, Soil, Insect etc. Solids: ~ 500 mg
Liquids: ~ 1,000 µL
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  • *1 LC-MS analysis (Dual Scan) can not be applicable for these sample types.
  • *2 Requires sample volume might be changed depending on analysis plan and study condition.
  • *3 Samples will be shipped to HMT America office in Boston from United States. If you are non-US researcher, please contact us.

Blood

Due to the less invasiveness and ease of sampling procedures, blood (plasma, serum and whole blood) is the most commonly used sample for biomarker discovery. Metabolome data from blood samples can vary greatly with different methods of sample preparation. HMT recommends the use of EDTA-treated plasma and whole blood samples because it does not undergo hemolysis and will provide data with small artificial metabolic changes. EDTA-treated plasma does not undergo hemolysis and will provide information about metabolites in pure plasma. In contrast, whole blood will offer information regarding metabolites in the whole blood, including blood cells. The required volume of blood sample is 50 ~ 100 µL for CE-MS analysis, and 200 – 500 µL for LC-MS analysis. The samples will be suspended in your laboratory, frozen and shipped to our office.

Urine

Urine, which reflects renal function, hepatic drug pharmacokinetics, and dietary habits, is a valuable sample for metabolome analysis. Advantages of urine are that it can be sampled in a non-invasive manner and that only deproteination by ultrafiltration, followed by dilution, is sufficient before urine can be subjected to CE-MS analysis. The required volume of urine sample is 50 ~ 100 µL for CE-MS analysis. The samples will be suspended in your laboratory, frozen and shipped to our office.

Animal or Plant Tissues

The tissue samples will be weighted, suspended in our transferring tube and frozen in your laboratory to prevent metabolite degradation. The required volume of tissue sample is 30 ~ 100 mg for CE-MS and LC-MS analysis, respectively. In HMT, metabolites are extracted according to established protocols selected by sample types, solvents, and other experimental conditions.

Selection of normalization methods is the starting point for the interpretation of results, but is also a critical task for metabolomics since no optimum methods that are applicable to all experimental systems are available. The present situation is that normalization methods are selected on a case-by-case basis according to experimental systems. Generally, normalization of data from tissue samples is based on tissue weight. However, normalization methods based on DNA levels or protein volume may be also used. As it is not easy to determine what types of normalization is the best, HMT recommends selecting different normalization methods for different experimental systems using statistical analysis approaches such as PCA on a case-by-case basis.

Cultured Cells

The culture cell samples will be extracted and inactivated in your laboratory to prevent metabolite degradation. The required volume of cell sample is 106 ~ 107, but might depends on cellular size. We recommends the use of our original protocol, which are reliable and easy work without any special reagent, but we can also treat frozen pellet or any other type of cellular samples. Because normal saline or buffered solutions such as phosphate buffered saline (PBS) contribute to the interference with CE-MS analysis, we will offer uses sugar alcohol solutions as wash solutions.

Normalization of data from cultures cells is usually based on cell count. However, normalization methods based on DNA levels and protein volumes may be also used when a comparison is made among cells with different volumes per cell.

Microorganisms

Single-cell microorganisms, such as E.coli, Yeast and Aspergillus, are used not only for basic biological research, but also in a variety of areas of study, including the production of useful materials such as proteins and fermented food. The samples will be extracted and inactivated in your laboratory to prevent metabolite degradation. The required volume of cell sample is 108 ~ 109. We recommends the use of our original protocol, which are reliable and easy work without any special reagent, but we can also treat frozen pellet or any other type of cellular samples.

Normalization of data from cultures cells is usually based on cell count. However, normalization methods based on DNA levels and protein volumes may be also used when a comparison is made among cells with different volumes per cell.

Others

We will provide appropriate sampling methods depending on research purpose. If your samples are not listed above, please contact us.

Please contact HMT for more information.