Biochemistry & Haematology
- Research assistant:
- Laboratory technicians:
Clinical chemistry analyses of plasma/serum and urine comprise of metabolites, ions, enzymes, and serological quantifications that could be used to assess metabolic and functional abnormalities of different organs of the body. Examination of whole blood for haematology may reveal pathologies or treatments that affect blood parameters. Both clinical chemistry and haematology are part of the core pipeline of the International Mouse Phenotyping Consortium (IMPC) with routine and standardized methods following IMPReSS guidelines.
In addition to IMPC compulsory parameters, the Biochemistry and Haematology Unit can further measure analytes within the capacity of the analyzers. We offer measurements of hormones, cytokines, serum and coagulation factors using immunoassays. As part of the metabolism platform of the CCP, we provide expert opinions and analysis for cell or tissue-specific characterization evaluating carbohydrate and lipid processing with established protocols.
Standard services Clinical Chemistry
The following analytes can be measured using the Beckman-Coulter AU480. The amount of samples needed depends on the desired parameters to be measured. Please refer to our attachment below for guidelines on sample collection and information regarding each assay and sample volume needed. For IMPC primary screens, Li-Heparin plasma is used. Some analytes can be quantified from urine and CSF. An additional 20 uL from each sample is needed to account for instrument dead volume and measurement of LIH (lipemia, icterus, & hemolysis assessment).
Calcium (Ca), Chloride (Cl), Inorganic Phosphorus (P), Iron (Fe), Magnesium (Mg), Potassium (K), Sodium (Na), Unsaturated-iron binding capacity (UIBC)
Albumin, Bilirubin (Conjugated &/or Total), Cholesterol (Total), HDL cholesterol (HDL),
LDL cholesterol (LDL), Creatinine, Ferritin, Fructosamines, Glucose, Glycerol, Lactate, Non-esterified Fatty Acids (NEFA), Total Protein, Transferrin, Triglycerides, Urea, Uric acid
Alpha-amylase, Alanine aminotransferase (ALT), Alkaline phosphatase (ALP), Aspartate aminotransferase (AST), Creatine Kinase (CpK), Hydroxybutyrate Dehydrogenase (HBDH), Lactate dehydrogenase (LDH), Lipase
Analytes can be analyzed as panels:
- Liver: Albumin, ALP, ALT, AST, Bilirubin, Total protein, Urea
- Kidney: Creatinine, Urea, Uric acid
- Pancreas: α-Amylase, Creatinine, Fructosamines, Glucose, Lipase
- Inflammation: ALP, ALT, AST, CK, Creatinine, Urea
- Lipid metabolism: Cholesterol (Total, HDL and LDL), Glycerol, Lipase, NEFA, Triglycerides
- Muscle (Cardiac) function: Na, K, Ca, CK, HBDH, LDH
- Bone: Ca, P, Albumin, ALP
- General: Na, K, Ca, P, Total protein, Albumin, ALP, AST, ALT, Urea, Creatinine, Cholesterol
- IMPC: ALT, Albumin, ALP, AST, Ca, Chloride, Creatinine, Glucose, HDL cholesterol, Fe, LDH, P, K, Na, Total bilirubin, Total cholesterol, Total protein, Triglycerides, Urea
Urine tests can be performed using the Beckman-Coulter AU480 and Siemens CLINITEK Advantus Urine Chemistry Analyzer.
With the Beckman-Coulter AU480 analyzer we can measure the following parameters quantitatively from the urine: urea, calcium, creatinine, glucose, magnesium, inorganic phosphate, uric acid, protein, microalbumin, potassium, sodium, and chloride. Note that for more in-depth assessment of renal function, urine collected over a period of 24 hours is preferable. Estimation of urinary protein excretion, glomerular filtration rate, and fractional excretion may require measurements of additional serum parameters.
The CLINITEK Advantus Urine Chemistry Analyzer is designed to read reagent strips for urinalysis and intended for the semi-quantitative and qualitative type of measurement of the following in the urine samples: Albumin, Bilirubin, Blood (occult), Creatinine, Glucose, Ketone, pH, Protein, Protein-to-Creatinine ratio, Albumin-to-Creatinine ratio, and Urobilinogen. Note that leukocyte esterase, specific gravity, and nitrite measurements using dipsticks might be unreliable for veterinary samples. The workflow includes automated quality checks and user-defined QC testing protocol.
Contact us for more information.
Please download the file below to check for other assays and reagents compatible with our analyzer:
Standard services Haematology
Basic haematological parameters from different animal species can be measured using the Mindray BC-5300 Vet. The instrument can measure at whole blood or pre-diluted modes. We require 25 uL of blood sample collected in EDTA-containing tubes. Assessing blood clotting and coagulation can be done using our automated Sysmex CA-560 system.
Complete Blood Count
The parameters that are measured are red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), and mean platelet volume (MPV).
Complete Blood Count with Differentials
This requires the same amount of EDTA-whole blood per sample and measures all the parameters above but also includes absolute and differential cell counts for neutrophils (Neu#, Neu%), lymphocytes (Lym#, Lym%), monocytes (Mon#, Mon%), eosinophils (Eos#, Eos%), and basophils (Bas#, Bas%).
Automated Blood Coagulation Analyzer
Our Sysmex Automated Blood Coagulation Analyzer is an in vitro diagnostic instrument capable of analysing up to 5 coagulation parameters, simultaneously. We can process parameters such as PT, APTT, Fibrinogen, Thrombin time (coagulation method), Antithrombin III (chromogenic method) or D-Dimer (immunoassay method). Anticoagulated plasma (in buffered sodium citrate) is needed for the tests. The plasma sample must be properly prepared from uncoagulated blood carefully extracted from the animal. Do not submit EDTA or heparin plasma, serum, serum separator or clot activator tubes.
Please click the link below to check for the different parameters that can be measured with the Sysmex® CA-560, the reagents, principle involved, and amount of sample and reagents needed.
Custom services Immunoassays
Using the Bio-Plex® 200 multiplex system, Epoch™ & Tecan Infinite F50 Spectrophotometers, we can measure multitude of biomolecules from a single sample. Samples that can be analyzed include serum/plasma, lavages, urine, milk, culture media, and cell/tissue culture supernatants. We currently offer different panels for multiplex assays and tested kits for individual analytes. For customization and complete lists of biomolecules that can be analysed in our multiplex system, please refer to the links.
Research Areas for Multiplexing:
Aging, Cardiovascular, Cell Signaling, Endocrinology, Epigenetics, Immunology, Metabolism, Neuroscience, Oncology, Osteology, and Toxicology
Aging, Acute phase proteins, Cardiovascular diseases, DNA damage, Signaling pathways & phosphoproteins, Pituitary hormones, Stress hormones, Sex & thyroid hormones, Histone post-translational modifications, Antibody isotyping, Cytokines/Chemokines, Diabetes, Metabolic hormones, Adipokines, Myokines, Neuropeptides, Amyloid beta, Angiogenesis, Immuno-oncology checkpoints, Bone metabolism, Vascular injury, Kidney toxicity, Liver injury
- Asthma/Airway Inflammation (Eotaxin, GM-CSF, IFNγ, IL-4, IL-5, IL-8, IL-10, IL-12, IL-13, IL-17, IL-18, TNFα)
- Cardiovascular Cytokine Profiling (CCL2 (MCP-1), CCL3 (MIP-1α), CCL4 (MIP-1β), CRP, CSF, CXCL16, Erythropoietin, FGF, Fractalkine (CX3CL1), G-CSF, GM-CSF, IFNγ, IL-1, IL-2, IL-5, IL-6, IL-8 (CXCL8), IL-10, IL-15, IL-18, M-CSF, PDGF, RANTES (CCL5), TNFα, VEGF)
- Cancer and Malignancy (Eotaxin, GM-CSF, IFNγ, IL-4, IL-5, IL-8, IL-10, IL-12, IL-13, IL-17, IL-18, TNFα)
- Rheumatoid Arthritis (IL-1β, IL-1RA, IL-6, IL-7, IL-10, IL-11, IL-12, IL-15, IL-17, IL-18, IL-23, MIP-3α, TGFβ, TNFα)
Pre-set and customized cytokines panels are available from Bio-Rad, ThermoFisher Scientific, Merck Millipore, and R&D Systems. Please download the PDF document to check for the comprehensive list of multiplex assays that the CCP can offer.
Insulin, Corticosterone, Leptin, Ghrelin, Hepcidin, TSH, T3, Thyroxine (T4), Testosterone, Estrogens, Progestins, PTH, FGF, Erythropoietin, Thrombopoietin, Pentraxin2/SAP, Cardiac Troponin-I, NT-proBNP, among others available upon request.
Custom services Assays for Metabolic Assessment on Different Tissues
In addition to standard services and immunoassays, the Unit offers tested and validated assays to further characterize carbohydrate and lipid metabolic states of different tissues like the liver, muscle, adipose, brain, and pancreas. Below is the partial list of services available. Please contact us for details regarding each assay and if you have further questions.
List of methods
- Lipid extraction and quantification from cells, tissues, or feces
- Intermediate term monitoring of glycemia using 1,5-Anhydroglucitol
- Isolation and quantification of liver glycogen
- Insulin ELISA for samples obtained during glucose tolerance test
- Fractionation of serum for lipoproteins using ultracentrifugation
- Other assays for cell & tissue lysates can be prepared and optimized on demand.
Custom services Clinical Pathology Panels
The Czech Centre for Phenogenomics offers multi-faceted phenotyping of animal models to characterized specific pathophysiology of interest. This involves clinical chemistry and haematology screenings together with analysis from other phenotyping units. In this customize option, combinations of analytes may be quantified from the samples depending on the research question. Examples of specific panels that the Biochemistry & Haematology Unit offers in coordination with other CCP units are:
Acute myocardial infarction
Cardiac troponin I, NT-proBNP, CPK, LDH, α-HBDH, Multiplex CVD Panel (see above)
RBC, HCT, Hb, RBC morphometry, WBC, Bilirubin, Fe, Ferritin, Mg, Transferrin, UIBC
Complete blood count (CBC), Uric acid, Total protein, Rheumatoid factor, Serum complement, Immunoglobulins, Serum antinuclear antibodies, Streptozyme and ASO tests
Core Tests: Albumin, ALP, ALT, Bilirubin, CRP, Creatinine, CBC, Potassium, Total protein, Sodium, Urea, Multiplex Airway Inflammation Panel (see above)
Extended Tests: Ferritin, cTNI, LDH, D-dimer, Procalcitonin
CBC+DIFF, Serum Bilirubin, ALP, AST, Amylase
Glucose (blood), Glucose (urine), Ketones (Acetoacetate, Acetone, β-Hydroxybutyrate), Potassium, Sodium, Chloride, Lactic acid, BUN, Phosphorus, HbA1c, Urine albumin, Multiplex Hormone/Diabetes Panel (see above)
Blood in stool, CBC, Urinalysis, BUN, Sodium, Potassium, Chloride, Total & direct bilirubin, AST, ALP, Albumin, Amylase
Prolactin, Growth hormone, ACTH, FSH, LH, TSH, T4, T3, Cortisol/Corticosterone, Testosterone, Progesterone, Estradiol
Standard Regulatory Toxicity Studies
Total Protein, Albumin, Cholesterol, Triglyceride, Glucose, ALT, AST, ALP, GGT, Total Bilirubin, Na, Cl, K, Ca, P, Urea, Creatinine, Multiplex Toxicity Panel (see above)
Other panels can be measured upon request.
Biochemistry and haematology unit was upgraded with the support from OP RDE project CZ.02.1.01/0.0/0.0/18_046/0015861 CCP Infrastructure Upgrade II