Thiazole orange: a new dye for reticulocyte analysis

Area of Science:

• Biomedical science
• Analytical chemistry
• Hematology

Background:

• Reticulocyte analysis is crucial for diagnosing and monitoring various anemias and bone marrow disorders.
• Current methods often rely on dyes with suboptimal excitation wavelengths, limiting their compatibility with standard flow cytometry equipment.
• Thioflavin T, a known dye, requires 457-nm excitation, prompting the search for a more versatile alternative.

Purpose of the Study:

• To identify and synthesize a novel fluorescent dye excitable at 488 nm for reticulocyte analysis.
• To evaluate the efficacy of newly synthesized dyes using fluorescence microscopy and flow cytometry.
• To establish a reliable and efficient method for reticulocyte enumeration.

Main Methods:

• Chemical synthesis of novel fluorescent dyes based on the thioflavin T structure.
• Evaluation of dye properties including quantum yield determination.
• Assessment of dye performance using fluorescence microscopy and fluorescence-activated cell cytometry (flow cytometry).
• Comparison of results with manual reticulocyte percentage determination.

Main Results:

• Several novel dyes were synthesized and tested.
• A dye named "thiazole orange" demonstrated superior performance.
• Thiazole orange is excitable at 488 nm, aligning with common flow cytometer lasers.
• Analysis of blood samples using thiazole orange showed a high correlation coefficient (0.97) with manual reticulocyte counts.

Conclusions:

• Thiazole orange is an effective 488-nm excitable fluorescent dye for reticulocyte analysis.
• This dye offers a significant improvement for flow cytometry-based reticulocyte enumeration.
• The high correlation with manual methods validates thiazole orange as a reliable diagnostic tool in hematology.