To quantify numbers of circulating reticulocytes, 5 [micro]l of blood was stained with 500 [micro]l thiazole orange
solution (Retic-Count, BD Biosciences) for 1 h at room temperature, centrifuged at 800x g for 5 min, and resuspended in 1% formaldehyde.
Kienast and Schmitz (5) were the first to describe a flow cytometric technique for analyzing retPLTs based on thiazole orange staining.
The Sapphire method is rather similar to the widely used flow cytometric method with thiazole orange, but it is fully automated and fully standardized regarding incubation time and concentration of the fluorescent RNA dye.
Flow cytometric analysis of thiazole orange uptake by platelets: a diagnostic aid in the evaluation of thrombocytopenic disorders.
The flow cytometric enumeration of reticulocytes uses fluorescent dyes that bind the residual RNA, such as thiazole orange (37, 38).
The use of thiazole orange, a fluorescent dye that binds RNA, allows immature platelets (also referred to as reticulated platelets) to be quantified (86-88).
Flow cytometric reticulocyte quantification using thiazole orange provides clinically useful reticulocyte maturity index.
Enrichment of reticulocytes was determined by means of thiazole orange labeling of reticulocytes in combination with flow cytometric analysis.
To 5 [micro]L of each combined cell fraction or whole blood, 1 mL of thiazole orange (Becton Dickinson) was added.