Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry
Abstract
Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry Linda Rodgers This protocol was adapted from “Flow Cytometry,” Chapter 16, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998. This threevolume set is now out of print; however, some of the microscopy methods were republished in Basic Methods in Microscopy, by David L. Spector and Robert D. Goldman. INTRODUCTION This protocol describes a method for quantitative measurement of DNA in tissue culture cells using either propidium iodide (PI) or DAPI staining followed by flow cytometry. PI can be excited at 488 nm by the argon-ion laser, the most commonly used laser in flow cytometry. Alternatively, DAPI is best excited by a high-power UV laser, which is less commonly available. After staining, cells can be stored frozen for later sorting or analysis. MATERIALS Reagents Boiled RNase A, 10 mg/ml Citrate buffer DMSO (Optional, see Step 5) DNA staining/lysis solution (For PI staining; see Step 2.) NST-DAPI buffer (For DAPI staining; see Step 2.) Equipment Flow cytometer Nylon mesh, 35-μm pore size (Small Parts, Inc.) Page 1 of 3 Propidium Iodide (PI) or DAPI Staining of Unfixed Tissue Culture Cells for Flow Cytometry
Related Papers
- → Analysis of Cell Cycle Regulation and Apoptosis Using Annexin-V and Propidium Iodide (PI) Staining(2023)2 cited
- → Effects of pH on a fluorogenic vital dyes assay (4′,6′-diamidino-2-phenyl-indole and propidium iodide) for Cryptosporidium sp. oocysts(1999)7 cited
- → Replacement of DAPI with propidium iodide could extend the utilisation of archival tissue samples for immunofluorescent techniques(2022)1 cited
- → Propidium Iodide (PI) or DAPI Staining of Unfixed Solid Tissues for Flow Cytometry(2006)
- Application of Ki67 and nuclear acid fluorescent dye co-staining in analysis of cell cycle(2016)