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PLoS One. 2015 May 11;10(5):e0125598. doi: 10.1371/journal.pone.0125598. eCollection 2015.

Proflavine Hemisulfate as a Fluorescent Contrast Agent for Point-of-Care Cytology.

Author information

1
Biomedical Engineering Department, University of Arkansas, Fayetteville, Arkansas 72701, United States of America.
2
10810 Executive Center Dr., Nephropath Ste. 100, Little Rock, Arkansas 72211, United States of America.

Abstract

Proflavine hemisulfate, an acridine-derived fluorescent dye, can be used as a rapid stain for cytologic examination of biological specimens. Proflavine fluorescently stains cell nuclei and cytoplasmic structures, owing to its small amphipathic structure and ability to intercalate DNA. In this manuscript, we demonstrated the use of proflavine as a rapid cytologic dye on a number of specimens, including normal exfoliated oral squamous cells, cultured human oral squamous carcinoma cells, and leukocytes derived from whole blood specimens using a custom-built, portable, LED-illuminated fluorescence microscope. No incubation time was needed after suspending cells in 0.01% (w/v) proflavine diluted in saline. Images of proflavine stained oral cells had clearly visible nuclei as well as granular cytoplasm, while stained leukocytes exhibited bright nuclei, and highlighted the multilobar nature of nuclei in neutrophils. We also demonstrated the utility of quantitative analysis of digital images of proflavine stained cells, which can be used to detect significant morphological differences between different cell types. Proflavine stained oral cells have well-defined nuclei and cell membranes which allowed for quantitative analysis of nuclear to cytoplasmic ratios, as well as image texture analysis to extract quantitative image features.

PMID:
25962131
PMCID:
PMC4427403
DOI:
10.1371/journal.pone.0125598
[Indexed for MEDLINE]
Free PMC Article

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