Periodic Acid-Schiff Staining of Cells: An In Vitro Technique to Detect Glycogen Levels in Peripheral Blood Mononuclear Cells

Abstract

Source: Tabatabaei Shafiei, M., et al. Detecting Glycogen in Peripheral Blood Mononuclear Cells with Periodic Acid Schiff Staining. J. Vis. Exp. (2014).

This video demonstrates the periodic acid-Schiff staining technique for assessing glycogen levels of peripheral blood mononuclear cells in vitro. The stain imparts a magenta color to intracellular glycogen granules, rendering them visible under a microscope.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. PBMC Isolation from Whole Blood

NOTE: Carry out this procedure in a biosafety cabinet using sterile technique and manufacturer-sterilized equipment.

1. Carefully pour 10-15 ml of whole blood from the heparinized (anti-coagulant) blood collection tubes into a 50 ml sterile conical tube. For minor blood spills, wipe with ddH₂O and 70% EtOH using cleaning tissue.
2. Dilute the blood in the conical tube to a 1:1....

Results

Figure 1: Step by step methodology of PAS staining on PBMC. (A) First, isolation of PBMC is achieved through ficoll gradient, the left panel shows the preparation before c.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Periodic Acid Shiff Kit	Sigma-Aldrich	395B	Bring to room temperature prior to use. Materials in this kit are toxic and harmful. Use caution.
α-Amylase from porcine pancreas	Sigma-Aldrich	A3176
Binocular Microscope	Carl Zeiss Microscopy	Axio Lab A0
Glycogen Assay Kit	Sigma-Aldrich	MAK016
Ficoll-Paque PLUS	VWR, GE Healthcare	17-1440-02	Nonionic synthetic polymer of sucrose.
Centrifuge			For PBMC isolation, swing buckets were used.