title: [Technical recommendations and best practice guidelines for May-Grünwald-Giemsa staining: literature review and insights from the quality assurance].
abstract: May-Grünwald-Giemsa (MGG) stain is a Romanowsky-type, polychromatic stain as those of Giemsa, Leishman and Wright. Apart being the reference method of haematology, it has become a routine stain of diagnostic cytopathology for the study of air-dried preparations (lymph node imprints, centrifuged body fluids and fine needle aspirations). In the context of their actions of promoting the principles of quality assurance in cytopathology, the French Association for Quality Assurance in Anatomic and Cytologic Pathology (AFAQAP) and the French Society of Clinical Cytology (SFCC) conducted a proficiency test on MGG stain in 2013. Results from the test, together with the review of literature data allow pre-analytical and analytical steps of MGG stain to be updated. Recommendations include rapid air-drying of cell preparations/imprints, fixation using either methanol or May-Grünwald alone for 3-10minutes, two-step staining: 50% May-Grünwald in buffer pH 6.8 v/v for 3-5minutes, followed by 10% buffered Giemsa solution for 10-30minutes, and running water for 1-3minutes. Quality evaluation must be performed on red blood cells (RBCs) and leukocytes, not on tumour cells. Under correct pH conditions, RBCs must appear pink-orange (acidophilic) or buff-coloured, neither green nor blue. Leukocyte cytoplasm must be almost transparent, with clearly delineated granules. However, staining may vary somewhat and testing is recommended for automated methods (slide stainers) which remain the standard for reproducibility. Though MGG stain remains the reference stain, Diff-Quik(®) stain can be used for the rapid evaluation of cell samples.
authors.affiliation: Centre de pathologie Est, hospices civils de Lyon, hôpitaux de Lyon, université Claude-Bernard Lyon 1, 59, boulevard Pinel, 69677 Bron cedex, France. Electronic address: eric.piaton@chu-lyon.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Piaton
authors.affiliation: Service d'anatomie et cytologie pathologiques, hôpital universitaire Necker-Enfants malades, université Paris Descartes, 75015 Paris, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Fabre
authors.affiliation: Laboratoire d'anatomopathologie, centre hospitalier René-Dubos, 95300 Cergy-Pontoise, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Goubin-Versini
authors.affiliation: Cabinet de pathologie, 67000 Strasbourg, France.
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Bretz-Grenier
authors.affiliation: Département d'anatomie et cytologie pathologiques, IUC Toulouse Oncopole, 31100 Toulouse, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Courtade-Saïdi
authors.affiliation: Unité de cytopathologie, institut Gustave-Roussy, 94805 Villejuif cedex, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Vincent
authors.affiliation: Service d'anatomie et cytologie pathologiques, hôpital Haut-Lévêque, CHU de Bordeaux, 33600 Pessac, France.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Belleannée
authors.affiliation: Centre de pathologie Est, hospices civils de Lyon, hôpitaux de Lyon, université Claude-Bernard Lyon 1, 59, boulevard Pinel, 69677 Bron cedex, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Thivolet
authors.affiliation: UF de cytologie pathologique, institut de biologie et de pathologie, CHU de Grenoble, 38700 La Tronche, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Boutonnat
authors.affiliation: Cabinet de pathologie, SCP des Drs Bloget et Declerck, 77210 Avon, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Debaque
authors.affiliation: Service d'anatomie et cytologie pathologiques, hôpital Tenon, GHU Est, 75020 Paris, France.
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Fleury-Feith
authors.affiliation: Unité de cytopathologie, institut Gustave-Roussy, 94805 Villejuif cedex, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Vielh
authors.affiliation: Service d'anatomie et cytologie pathologiques, université Paris Descartes, hôpitaux universitaires Paris Centre, site Cochin, AP-HP, 75014 Paris, France.
authors.forename: Béatrix
authors.initials: B
authors.lastname: Cochand-Priollet
authors.affiliation: Département de pathologie, hôpital de Hautepierre, CHU de Strasbourg, 67200 Strasbourg, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Egelé
authors.affiliation: Département de pathologie, hôpital de Hautepierre, CHU de Strasbourg, 67200 Strasbourg, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Bellocq
authors.affiliation: Service d'anatomie et cytologie pathologiques, hôpital Pasteur, CHU de Nice, 06001 Nice cedex 1, France.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Michiels
mesh_terms: D001331:Automation; D001399:Azure Stains; D003585:Cell Biology / Q000458:organization & administration; D004396:Coloring Agents / Q000737:chemistry; D003581:Cytodiagnosis / Q000379:methods / Q000592:standards*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D004912:Erythrocytes / Q000648:ultrastructure; D005602:France; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007962:Leukocytes / Q000648:ultrastructure; D008751:Methylene Blue / Q000737:chemistry; D015388:Organelles / Q000648:ultrastructure; D017410:Practice Guidelines as Topic; D011785:Quality Assurance, Health Care; D015203:Reproducibility of Results; D012958:Societies, Scientific; D013194:Staining and Labeling / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D016707:Tissue Fixation / Q000379:methods; D014966:Xanthenes
chemical_list: D001399:Azure Stains; D004396:Coloring Agents; C067461:Diff Quik; C043260:May-Grunwald Giemsa; D014966:Xanthenes; D008751:Methylene Blue; D004801:Eosine Yellowish-(YS)
keywords: Assurance qualité; Coloration; Cytologie; Cytopathology; Giemsa; May-Grünwald; May–Grünwald; Quality assurance; Romanowsky; Stain; Standardisation; Standardization; Wright