Fig 1.
Custom epi-fluorescence microscopy system.
Comprises a Flea 3 USB camera, 525/40nm bandpass emission filter, 475nm dichroic filter set, 455nm LED light source, 40x Nikon Plan Achromat air objective and two linear translation stages.
Fig 2.
Illustration of manually-segmented oral epithelial cells stained with proflavine.
Top row: Normal exfoliated oral epithelial cells. Bottom row: CAL 27 oral squamous carcinoma cells. Scale bars = 10μm.
Fig 3.
Visual diagnosis comparison of Papanicolaou and proflavine-stained normal oral cells and CAL 27 cell line.
First row: Papanicolaou stained normal oral cells. Second row: Papanicolaou stained CAL 27 cells. Third row: Fluorescent images of proflavine stained normal oral cells. Fourth row: Fluorescent images of proflavine stained CAL 27 cells. Scale bars = 10 μm.
Fig 4.
Comparison of cellular features seen in Papanicolaou (top row) and proflavine (bottom row) stained cells.
Left column, oral squamous epithelial cells. Right column, Cal 27 squamous carcinoma cells. NU: Nucleus, NO: Nucleolus, KH: Keratohyalin granule, GZ: Golgi zone. Scale bars = 10 μm.
Fig 5.
Giemsa (top row) and proflavine (bottom row) stained leukocytes.
First row, Giemsa stained neutrophil (A), monocyte (B), lymphocyte (C). Second row, proflavine stained neutrophil (D), monocyte (E), lymphocyte (F). Images were captured at 60X. Fluorescence images were normalized as described in the Methods section. Scale bars = 10μm.
Fig 6.
Bar plot comparing nuclear to cytoplasmic ratios for normal and CAL 27 cells.
P value << 0.001 with Student T test. Error bars denote standard deviation.
Fig 7.
Bar plot comparing features between normal and CAL 27 cells.
For both plots, p value << 0.001 with Student T test. 100 cells per group. Error bars denote standard deviation.
Table 1.
Comparison of cost and time for pathology stains.
Table 2.
Comparison of proflavine and PAP stained oral cells.
Table 3.
Comparison of proflavine and Giemsa stained leukocytes.