In addition, a recently developed methodology is applied which uses induced circular dichroism spectral profiles of G-quadruplex-ligand (Thiazole Orange) complexes to determine if G-rich sequences can adopt G-quadruplex structure.
Thiazole Orange was purchased form Sigma-Aldrich (cat.
The organism, suspended in physiological saline, was subjected for 5 min to either heat (55 or 60[degrees]C) or ultrasound (20 kHz), or to a combination of both, and analysed for viable, membrane-injured and dead cells using thiazole orange
(TO) and propidium iodide (PI).
The principle depends on enumeration of CD61 and thiazole orange positive cells.
Then one ml of thiazole orange was added to the previous tube, capped and mixed gently then incubated in the dark at room temperature for 15 minutes.
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.
Reticulocyte count using thiazole orange. A flow cytometry method.
In previous reports, different dyes such as Hoechst 33258 , acridine orange , thiazole orange
, or hydroethidine  were used to determine parasitemia in cultures of Plasmodium falciparum by FACS.
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.
The flow cytometric enumeration of reticulocytes uses fluorescent dyes that bind the residual RNA, such as thiazole orange
Enrichment of reticulocytes was determined by means of thiazole orange
labeling of reticulocytes in combination with flow cytometric analysis.