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1. (WO2019007925) METHOD FOR THE MULTI-PARAMETRIC ANALYSIS OF PROGRAMMED CELL DEATH BY FLOW CYTOMETRY
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Pub. No.: WO/2019/007925 International Application No.: PCT/EP2018/067875
Publication Date: 10.01.2019 International Filing Date: 03.07.2018
IPC:
G01N 33/58 (2006.01) ,G01N 33/92 (2006.01)
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
58
involving labelled substances
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
92
involving lipids, e.g. cholesterol
Applicants:
ISTITUTO SUPERIORE DI SANITA' [IT/IT]; Viale Regina Elena, 299 00161 Roma, IT
Inventors:
MARIOTTI, Sabrina; IT
NISINI, Roberto; IT
Agent:
VALENZA, Silvia; IT
GERVASI, Gemma; IT
PALLINI GERVASI, Diego; IT
CATTANEO, Elisabetta; IT
PALLADINO, Massimo; IT
ZACCARO, Elisabetta; IT
COPPO, Alessandro; IT
CELONA, Antonio; IT
DE BORTOLI, Tiziano; IT
BRIGHENTI, Livio; IT
SAVI, Massimiliano; IT
TORTI, Carlo Maria Emilio; IT
CINQUANTINI, Bruno; IT
CONTADIN, Giorgio; IT
Priority Data:
10201700007406003.07.2017IT
Title (EN) METHOD FOR THE MULTI-PARAMETRIC ANALYSIS OF PROGRAMMED CELL DEATH BY FLOW CYTOMETRY
(FR) PROCÉDÉ D'ANALYSE MULTIPARAMÉTRIQUE DE MORT CELLULAIRE PROGRAMMÉE PAR CYTOMÉTRIE EN FLUX
Abstract:
(EN) The present invention discloses a simple method (Fixed Apoptotic/Necrotic cells test: FAN) that is not influenced by fixation or permeabilization of cells, thus permitting complete multi-parametric flow cytometric analyses of cells undergoing programmed cell death even several days after the staining. FAN uses reagents that maintain their binding and fluorescence characteristics after fixation and/or permeabilization: a fluorochrome-labelled anti-phosphatidylserine antibody and a choice of fluorescent amine-binding dyes. FAN represents a new tool to study cells undergoing apoptosis and/or necrosis with exclusive characteristics, which make it the unique flow cytometry method in the study of infections when fixation procedures are required for safe sample handling.
(FR) La présente invention concerne un procédé simple (test de cellules apoptotiques/nécrotiques fixes : FAN) qui n'est pas influencé par la fixation ou la perméabilisation de cellules, permettant ainsi d'effectuer des analyses cytométriques en flux multiparamétriques complètes de cellules subissant une mort cellulaire programmée même plusieurs jours après la coloration. Le FAN utilise des réactifs qui conservent leurs caractéristiques de liaison et de fluorescence après fixation et/ou perméabilisation : un anticorps anti-phosphatidylsérine marqué par un fluorochrome et un choix de colorants de liaison à une amine fluorescente. Le FAN représente un nouvel outil permettant d'étudier des cellules subissant une apoptose et/ou une nécrose à l'aide de caractéristiques exclusives, ce qui en fait le procédé de cytométrie en flux unique dans l'étude d'infections lorsque des procédures de fixation sont exigées pour une manipulation d'échantillon sûre.
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)