- A second kind of techniques requires short incubation (hours) or long incubation (days) of cells; the analysis is performed either on the cells or on the soluble molecules released in the culture supernatant.

- The third type is image analysis.

- The fourth is molecular analysis.

Quantitation of cytokine secretion in biological fluids and in the culture supernatants of stimulated immune cells, by ELISA and flowcytometry

• TNF-, IFN-, IL-13, IL-10, IL-4, IL-2 etc

Immunophenotyping of circulating lymphocytes, normal or leukemic, from bone marrow aspirate, pathological inflammatory exudates or bronchoalveolar liquid by flow cytometry

• gradient density separation of cells

• labelling with fluorescent antibodies (1-4-8 colors)

Flowcytometric evaluation of intracellular cytokine panel induced by various diseases

• cell isolation

• antigenic or mitogenic stimulation of cells in short-term or long-term cultures

• intracellular labelling for IFN-, IL-2, TNF-/, IL-5, IL-10, IL-13

• flow cytometric analysis

Cell proliferation measurement following in vitro antigenic stimulation

• PBMC density gradient separation

• carbofluorescein diacetat succinimidil ester (CFSE) staining

• cell culture, in vitro mitogenic stimulation

• surface staining with fluorescent monoclonal antibodies and flow-cytometry

Cell cycle measurement for evaluation of neoplasic status

• tissue disruption for generating cell suspensions

• staining with propidium iodide

• flow cytometry

Image analysis

(Immunohistochemistry/ Immunochemistry/Immunofluorescence/FISH and TissueFaxs)

• processing of bioptic material

• incubation with different monoclonal

Phenotyping for HLA

• Molecular biology

Constitutional chromosome analysis

• Lymphocyte culture

• Syncronised lymphocyte culture

• Metaphasic chromosome labeling

• Prometaphasic chromosome labeling

• Chromosome analysis and imaging

• Karyotyping and evaluation

FISH for microdeletion and other genetic disordes

(Wolf-Hirschorn, Williams, Preder Willi/Angelmann, Velo-Cardio-Facial, Down syndromes, X,Y aneuploidies)

Chromosome analysis of bone marrow samples (malignant hemopathies)

Molecular Biology

Multiplex PCR for BRCA investigation

PCR for fragile X, Y microdeletion, fetal DNA in maternal plasma, FLT3 mutation in LAM

RT-PCR (Real Time PCR): Cytokine polymorphism IL-1A, IL-1B, IL-6, TNF, IL-4, IL-4delta2, IFNgamma; BCR/ABL translocation; fetal DNA in maternal plasma

MLPA for BRCA mutations, mentally retarded child.

Sequencing for BRCA mutations analysis

Serological tests for identification and quantitation of tumor markers

• ELISA quantitation of tumor markers

Serological tests for the identification of viral markers

• ELISA identification of viral antigens in sera or specific antibodies

Quantitation of autoantibodies for autoimmunity diagnosis

ELISA and Chemiluminescence

Quantitation of circulating IgM, IgG, IgA,IgE CRP, C3 fraction and circulating immune complexes

Nephelometry, Chemiluminescence