title: RCL2, a new fixative, preserves morphology and nucleic acid integrity in paraffin-embedded breast carcinoma and microdissected breast tumor cells.
abstract: Methacarn and RCL2, a new noncrosslinking fixative, were compared to formalin-fixed or frozen tissue samples of the same invasive breast carcinoma and were evaluated for their effects on tissue morphology and immunohistochemistry as well as DNA and RNA integrity. The histomorphology of methacarn- or RCL2-fixed paraffin-embedded tumors was similar to that observed with the matched formalin-fixed tissues. Immunohistochemistry using various antibodies showed comparable results with either fixative, leading to accurate breast tumor diagnosis and determination of estrogen and progesterone receptors, and HER2 status. Methacarn and RCL2 fixation preserved DNA integrity as demonstrated by successful amplification and sequencing of large DNA amplicons. Similarly, high-quality RNA could be extracted from methacarn- or RCL2-fixed paraffin-embedded MCF-7 cells, whole breast tumor tissues, or microdissected breast tumor cells, as assessed by electropherogram profiles and real-time reverse transcriptase-polymerase chain reaction quantification of various genes. Moreover, tissue morphology and RNA integrity were preserved after 8 months of storage. Altogether, these results indicate that methacarn, as previously shown, and RCL2, a promising new fixative, have great potential for performing both morphological and molecular analyses on the same fixed tissue sample, even after laser-capture microdissection, and can open new doors for investigating small target lesions such as premalignant breast lesions.
authors.affiliation: Laboratoire d'Anatomie Pathologique, Centre Hospitalier Universitaire de Montpellier, Hôpital Arnaud de Villeneuve, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Delfour
authors.forename: Pascal
authors.initials: P
authors.lastname: Roger
authors.forename: Caroline
authors.initials: C
authors.lastname: Bret
authors.forename: Marie-Laurence
authors.initials: ML
authors.lastname: Berthe
authors.forename: Philippe
authors.initials: P
authors.lastname: Rochaix
authors.forename: Nicolas
authors.initials: N
authors.lastname: Kalfa
authors.forename: Pierre
authors.initials: P
authors.lastname: Raynaud
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bibeau
authors.forename: Thierry
authors.initials: T
authors.lastname: Maudelonde
authors.forename: Nathalie
authors.initials: N
authors.lastname: Boulle
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D045744:Cell Line, Tumor; D002877:Chromosomes, Human / Q000235:genetics; D004247:DNA / Q000235:genetics; D005404:Fixatives; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009696:Nucleic Acids / Q000235:genetics*; D016612:Paraffin Embedding; D012313:RNA / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics
chemical_list: D004247:DNA; D005404:Fixatives; D009696:Nucleic Acids; D012313:RNA; D018719:Erb-b2 Receptor Tyrosine Kinases