Flow cytometry (Figure
10) is a technique used for measuring the chemical or physical characteristics of cells or particles. The cells or particles of interest are suspended in solution, labelled with fluorescent markers, and run through a cell analyser ‐ a flow cytometer instrument. The cells or particles flow through a small tube one at a time, and a laser is focused on them, scattering light and fluorescence of different wavelengths then recorded by the flow cytometer. Data are presented on a computer as either single parameter histograms or two‐parameter plots called cytograms.
The protocol describing handling and preparation of the blood samples before running on the flow cytometer is described thoroughly in papers II, III and IV (Appendix
A page 35).
We used a
BD FACSCantoII flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), and to analyse the data, we used the software
Kaluza Analysis (
Kaluza Analysis version 2.1; Beckman Coulter, Inc., Pasadena, CA, USA). For measuring white blood cell count (WBC) to obtain 1.0 x 10
6 leukocytes in the test tubes, we used a
Sysmex KX‐21NTM (Sysmex Corporation, Kobe, Japan).
Table
5 is an overview of the monoclonal antibodies used for flow cytometry. A gating strategy for the flow cytometric analyses is shown in paper IV (page 35).
, & Liisborg C. (2022). Age‐related macular degeneration and myeloproliferative neoplasms – A common pathway. Acta Ophthalmologica, 100(Suppl 271), 3-35.