Periodic Acid-Schiff Staining of Cells: An In Vitro Technique to Detect Glycogen Levels in Peripheral Blood Mononuclear Cells
Published: April 30, 2023
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. Carefully pour 10-15 ml of whole blood from the heparinized (anti-coagulant) blood collection tubes into a 50 ml sterile conica…
Representative 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 centrifugation, the right panel shows it after centrifugation where the buffy coat containing the PBMC is observed in the center of the tube. (B) Isolated PBMCs are fixed onto the slide using formalin-ethanol…
開示
The authors have nothing to disclose.
Materials
| 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. |
