Bringing the EuroFlow Concept

Bringing the EuroFlow Concept

Cytognos - EuroFlow Supporting Company Company Overview Cytognos provides through worldwide distribution a broad range of reagents and software for flow cytometry applications in clinical laboratories. Cytognos, EuroFlow supporting company, plays the role of providing solutions for flow cytometry. That purpose led to the release of the EuroFlow product line: Screening Tubes, Infinicyt Software and Specialized Technical Support. Product Line Screening Tubes 8-color immunostaining for fast and easy flow cytometric analysis. Cytognos screening tubes (LST-11, ALOT-7 and SST) include markers that have been selected to allow accurate identification of cell populations with a single tube. Infinicyt Software Easy handling of large datasets and integration of 8-color staining into a single multicolor file. Infinicyt facilitates interpretation and automated pattern recognition of normal, reactive, and aberrant leucocyte populations. Specialized Technical Support Cytognos technical specialists are available to provide fieldor email-based assistance and support. Cytognos specialists are highly qualified personnel who can help on: Calibration and compensation protocols. Preparation and staining protocols. Infinicyt analysis. EuroFlow Consortium EuroFlow is a scientific consortium composed of research groups and companies aiming for the development and standardization of fast, accurate, and highly sensitive flow cytometry tests for diagnosis and prognostic classification of hematological malignancies as well as for evaluation of treatment effectiveness during follow-up (1). EuroFlow antibody panels are unique due to its multicenter validation process performed by several laboratories in successive evaluation rounds and large number of patients (1,2). EuroFlow 8-color antibody panels and EuroFlow standard operating procedures (SOPs) have been designed to be used in combination with Infinicyt software for multivariate data analysis (1-6). The publications are available at www.euroflow.org. Contact us at support@cytognos.com and ask for specialized technical support

EuroFlow Screening Tubes EuroFlow screening tubes are intended for a rapid differential identification of all relevant cell subsets. Subsequent to initial sample screening, specific antibody panels are applied for further characterization and classification of the suspected cells. Novel fluorochromes Cytognos offers two new fluorochromes, OC515 and APC-C750, that have been conjugated with different antibodies to complement the EuroFlow panels. OC515 is compatible to Pacific Orange and APC-C750 to APC-H7. These new fluorochromes reach EuroFlow high standards in relation to stability, brightness, limited spectral overlap and compensation requirements. Validated antibodies Cytognos offers a list of antibodies which can be used as a starting point for all EuroFlow antibody panels. Ask for a sample at admin@cytognos.com

Screening Kits LST-11 KIT Lymphoid Screening Tube The LST-11 kit is designed for detecting aberrant mature lymphocyte populations (B, T and NK lineage). It is used to evaluate several clinical manifestations such as peripheral blood lymphocytosis, bone marrow lymphoid infiltrates, enlargement of one or multiple other tissues (e.g. lymphadenopathy, splenomegaly or other organomegalies) or emergence of a serum monoclonal component. PacB OC515 FITC PE PerCP-Cyanine5.5 PE-Cyanine7 APC APC-C750 CD20 CD4 CD45 CD8 SmIgλ CD56 SmIgκ CD5 CD19 SmCD3 CD38 Pathological peripheral blood sample stained with LST-11 kit. Clonal Ig κ B-cell population (purple), remaining normal B-cell population (blue) and T-cell population (green); remaining events are shown in grey in CD45:OC515 vs. SSC dot plot. Normal B cells Clonal B cells T cells Outstanding Features (LST and SST) Ready-to-use EuroFlow combination Lyophilized format Selected fluorochromes Time saving Data accuracy Reliable results High stability Fewer storage requirements Exceptional brightness Minimum spectral overlap Give an extra value to your laboratory work flow with our EuroFlow product line Contact us at support@cytognos.com and ask for specialized technical support

SST KIT Small Sample Screening Tube The SST kit is an enhanced version of the EuroFlow LST. It is optimized for the analysis of low cellularity samples such as cerebrospinal fluid, vitreous humour, fine needle aspirates, etc. PacB OC515 FITC PE PerCP-Cyanine5.5 PE-Cyanine7 APC APC-C750 CD20 CD45 CD8 SmIgλ CD56 SmIgκ CD4 CD19 SmCD3 CD14 CD38 Pathological cerebrospinal fluid stained with SST kit. Clonal Ig κ B-cell population (purple) and T-cell population (green). Clonal B cells T cells Marker CD3 CD4 CD5 CD8 CD14 CD19 CD20 CD38 CD45 CD56 SmIgκ SmIgλ Reactivity Mature T cells T helper cells, monocytes, macrophages and dendritic cells Mature T cells, subpopulation of B cells T cytotoxic cells, subpopulation of NK cells Monocytes B cells (precursors to plasma cells) Mature B cells (except plasma cells) Plasma cells, activated T cells, B cell precursors Leucocytes NK cells, subpopulation of T cells Kappa light chain (B cells) Lambda light chain (B cells) Ask for a sample at admin@cytognos.com

Screening Kits ALOT-7 KIT Acute Leukemia Orientation Tube The ALOT-7 kit is designed for initial assessment of the nature of immature populations of hematopoietic cells in samples with suspicion of acute leukemia (B- or T-lymphoid vs. non-lymphoid lineage or mixed phenotype). PacB OC515 FITC PE PerCP-Cy5.5 PE-Cyanine7 APC APC-C750 CyCD3 CD45 CyMPO CyCD79a - CD19 CD7 SmCD3 Pathological bone marrow sample stained with ALOT-7 kit. B lineage blast cells (purple), remaining normal B-cell population (blue), normal T-cell population (yellow) and neutrophils (green). Normal B cells Blast cells T cells Neutrophils Euroflow Panels The ALOT-7 kit, in combination with CD34- PerCP-Cy5.5, can orientate with exactitude further analysis of acute leukemia samples. Together with BCP-ALL, B-CLPD, and AML/MDS panels, the complete detection or exclusion of virtually all types of hematological diseases can be achieved. Infinicyt Compass tool helps to orientate later analysis by comparison of pathological samples with the ALOT screening tube database. Outstanding Features Cytoplasmic antibodies in a single vial EuroFlow combination Selected fluorochromes Marker CyMPO CD3 CD7 CyCD79a CD19 CD45 Reactivity Time saving Data accuracy Reliable results Exceptional brightness Minimum spectral overlap Myeloid lineage Immature T cells (Cy) and mature T cells (Sm) Mature T cells, NK cells, hematopoietic cells B cell precursors B cells (precursors to plasma cells) Leucocytes Contact us at support@cytognos.com and ask for specialized technical support

Validated antibodies Validated Antibodies Cytognos has included remarkable antibodies within EuroFlow reference antibody list. Consult www.euroflow.org to obtain the last updated list. Markers CD8-FITC CD38-Pure CD38-FITC CD45-OC515 CD56-PE cycd79a-pe SmIgΚ-PE SmIgλ-FITC cytcrβ-apc Application in EuroFlow LST, SST and B-CLPD PCD PCD ALOT, LST, SST, AML, B-CLPD, BCP-ALL, NK-CLPD, T-ALL and T-CLPD LST, SST, PCD, AML and B-CLPD ALOT LST, SST, PCD, B-CLPD and BCP-ALL LST, SST, PCD, B-CLPD and BCP-ALL T-ALL Reference antibodies Alternative antibodies Technical support Cytognos technical specialists are highly qualified personnel who have been trained to provide specialized support for EuroFlow product line to our customers. Our specialists are available to provide field- or email-based assistance on: EuroFlow Standard Operating Protocol for instrument setup and compensation. EuroFlow Standard Operating Procedures for sample preparation and staining. Analysis of flow cytometric data with Infinicyt software. Ask for a sample at admin@cytognos.com

Literature 1 Van Dongen JJM, Orfao A. EuroFlow: Resetting leukemia and lymphoma immunophenotyping. Basis for Companion Diagnostics in personalized medicine. LEUKEMIA 2012, in press. 2 Van Dongen JJM, Lhermitte L, Böttcher S, Almeida J, van der Velden VHJ, Flores-Montero J, Rawstron A, Asnafi V, Lécrevisse Q, Lucio P, Mejstrikova E, Szczepanski T, Kalina T, de Tute R, Brüggemann M, Sedek L, Cullen M, Langerak AW, Mendonça A, Macintyre E, Martin-Ayuso M, Hrusak O, Vidriales MB, Orfao A. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. LEUKEMIA 2012, in press. 3 Kalina T, Flores-Montero J, van der Velden VHJ, Martin-Ayuso M, Böttcher S, Ritgen M, Almeida J, Lhermitte L, Asnafi V, Mendonça A, de Tute R, Cullen M, Sedek L, Vidriales MB, Pérez JJ, te Marvelde JG, Mejstrikova E, Hrusak O, Szczepanski T, van Dongen JJM, Orfao A. EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols. LEUKEMIA 2012, in press. 4 Pedreira CE, Costa ES, Barrena S, Lecrevisse Q, Almeida J, van Dongen JJM, Orfao A. Generation of flow cytometry data files with a potentially infinite number of dimensions. CYTOMETRY, PART A, 2008, 73A: 834 846. 5 Pedreira CE, Costa ES, Almeida J, Fernández C, Quijano S, Flores J, Barrena S, Lecrevisse Q, van Dongen JJM, Orfao A. A probabilistic approach for the evaluation of minimal residual disease by multiparameter flow cytometry in leukemic B-cell chronic lymphoproliferative disorders. CYTOMETRY, PART A, 2008, 73A: 1141 1150. 6 Costa ES, Peres RT, Almeida J, Lécrevisse Q, Arroyo ME, Teodósio C, Pedreira CE, van Dongen JJ, Orfao A. Harmonization of light scatter and fluorescence flow cytometry profiles obtained after staining peripheral blood leucocytes for cell surface-only versus intracellular antigens with the Fix & Perm reagents. CYTOMETRY, PART B, 2010, 78B: 11 20. More publications related to EuroFlow consortium and EuroFlow product line can be found at: http://www.euroflow.org/publications.php and http://www.infinicyt.com/index.php/publications. Spain Pol. La Serna, Nave 9 37900 Santa Marta de Tormes Salamanca 34 923 125 067 Contact information The Netherlands Erasmus MC (Faculteitsgebouw) Department of Immunology Room Ee 818 Dr. Molewaterplein 50 3015 GE Rotterdam Consult which Cytognos distributor is in your area at www.cytognos.com cytognos Flow Cytometry Solutions Design and development of reagents, software and techniques that provide innovative solutions in the flow cytometry field. www.cytognos.com admin@cytognos.com support@cytognos.com www.infinicyt.com Follow us