title: Detecting glycogen in peripheral blood mononuclear cells with periodic acid schiff staining.
abstract: Periodic acid Schiff (PAS) staining is an immunohistochemical technique used on muscle biopsies and as a diagnostic tool for blood samples. Polysaccharides such as glycogen, glycoproteins, and glycolipids stain bright magenta making it easy to enumerate positive and negative cells within the tissue. In muscle cells PAS staining is used to determine the glycogen content in different types of muscle cells, while in blood cell samples PAS staining has been explored as a diagnostic tool for a variety of conditions. Blood contains a proportion of white blood cells that belong to the immune system. The notion that cells of the immune system possess glycogen and use it as an energy source has not been widely explored. Here, we describe an adapted version of the PAS staining protocol that can be applied on peripheral blood mononuclear immune cells from human venous blood. Small cells with PAS-positive granules and larger cells with diffuse PAS staining were observed. Treatment of samples with amylase abrogates these patterns confirming the specificity of the stain. An alternate technique based on enzymatic digestion confirmed the presence and amount of glycogen in the samples. This protocol is useful for hematologists or immunologists studying polysaccharide content in blood-derived lymphocytes.
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University.
authors.forename: Mahdieh
authors.initials: M
authors.lastname: Tabatabaei Shafiei
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University.
authors.forename: Catalina M
authors.initials: CM
authors.lastname: Carvajal Gonczi
authors.affiliation: Department of Chemistry and Biochemistry, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University.
authors.forename: Mohammed Samiur
authors.initials: MS
authors.lastname: Rahman
authors.affiliation: Department of Exercise Science, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University.
authors.forename: Ashley
authors.initials: A
authors.lastname: East
authors.affiliation: Department of Exercise Science, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University.
authors.forename: Jonathan
authors.initials: J
authors.lastname: François
authors.affiliation: Department of Biology, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University; Department of Exercise Science, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University; peter.darlington@concordia.ca.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Darlington
mesh_terms: D000681:Amylases / Q000737:chemistry; D006003:Glycogen / Q000032:analysis / Q000097:blood*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000378:metabolism; D010503:Periodic Acid-Schiff Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D006003:Glycogen; D000681:Amylases