title: A new and improved "quick-hot Gram-chromotrope" technique that differentially stains microsporidian spores in clinical samples, including paraffin-embedded tissue sections.
abstract: OBJECTIVE
  This report describes a new and improved "quick-hot Gram-chromotrope" staining technique that detects microsporidian spores in clinical specimens, such as stool, urine, saliva, nasopharyngeal fluid, and bronchoalveolar lavage samples, as well as in formalin-fixed and paraffin-embedded tissue sections.
  
  
  METHODS
  In this procedure, the samples are stained in heated (50 degrees C to 55 degrees C) solutions of crystal violet and iodine used in Gram's stain, followed by a modified chromotrope solution (heated to 50 degrees C to 55 degrees C). The modified stain is composed of chromotrope 2R (1%), fast green (0.15%), and phosphotungstic acid (0.25%).
  
  
  RESULTS
  With this stain and the new protocol, microsporidian spores are stained dark violet against a pale green background, and the total staining time is shortened to 5 minutes.
  
  
  CONCLUSIONS
  This new technique is fast, reliable, and simple. It can be easily adapted for use in clinical laboratories.
authors.affiliation: Division of Parasitic Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30341-3724, USA.
authors.forename: H
authors.initials: H
authors.lastname: Moura
authors.forename: D A
authors.initials: DA
authors.lastname: Schwartz
authors.forename: F
authors.initials: F
authors.lastname: Bornay-Llinares
authors.forename: F C
authors.initials: FC
authors.lastname: Sodré
authors.forename: S
authors.initials: S
authors.lastname: Wallace
authors.forename: G S
authors.initials: GS
authors.lastname: Visvesvara
mesh_terms: D000818:Animals; D001826:Body Fluids / Q000469:parasitology; D018929:Cell Culture Techniques; D005243:Feces / Q000469:parasitology; D005840:Gentian Violet / Q000737:chemistry*; D006801:Humans; D007411:Intestinal Diseases, Parasitic / Q000175:diagnosis*; D016816:Microsporida / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D016881:Microsporidiosis / Q000175:diagnosis*; D016612:Paraffin Embedding; D010619:Phenazines / Q000737:chemistry*; D013170:Spores / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: C035863:Gram's stain; D010619:Phenazines; D005840:Gentian Violet