Confocal microscopy and exfoliative cytology

Reddy, Shyam Prasad; Ramani, Pratibha; Nainani, Purshotam

doi:10.4103/0973-029x.119746

Cited by 7 publications

(3 citation statements)

References 9 publications

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“…Proflavine, as an aminoacridine derivative, is a promising contrast agent for point-of-care cytology due to its rapid uptake, highly simplified staining procedure, and robust and consistent staining of cellular structures. Other aminoacridine derivatives, such as acridine orange- with a peak excitation wavelength of 450 and emission of 650- have also been widely studied as a nucleic acid stain [ 37 ]. Acridine orange has also been studied for point-of-care diagnostics, such as malaria, due to its rapid and selective staining and high contrast images of intracellular plamsmodium parasites [ 38 , 39 ].…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2015

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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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2015

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“…The confocal Raman microscope has been widely applied in biology [29][30][31][32] as well as in material science [33][34][35]. The objective of this paper is to investigate the application of this ubiquitous and common instrument for multicomponent Raman analysis.…”

Section: A Confocal Raman Microscopymentioning

confidence: 99%

Multicomponent analysis using a confocal Raman microscope

et al. 2018

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Measuring the concentration of multiple chemical components in a low-volume aqueous mixture by Raman spectroscopy has received significant interest in the literature. All of the contributions to date focus on the design of optical systems that facilitate the recording of spectra with high signal-to-noise ratio by collecting as many Raman scattered photons as possible. In this study, the confocal Raman microscope setup is investigated for multicomponent analysis. Partial least-squares regression is used to quantify physiologically relevant aqueous mixtures of glucose, lactic acid, and urea. The predicted error is 17.81 mg/dL for glucose, 10.6 mg/dL for lactic acid, and 7.6 mg/dL for urea, although this can be improved with increased acquisition times. A theoretical analysis of the method is proposed, which relates the numerical aperture and the magnification of the microscope objective, as well as the confocal pinhole size, to the performance of the technique.

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“…Compared with all of the set-ups mentioned in Section 2 for which the application range is essentially limited to recording Raman spectra from bulk (aqueous) samples, confocal Raman microscopy provides for a diversity in application including neurophysiology, cytology, material science, etc. [26][27][28] The simple design includes a pinhole between the sample and the detector, the benefit of which is to reduce the unnecessary background scattering from optical elements and from the substrate, as well as to improve the three dimensional spatial resolution to approximately one micrometer in all three dimensions. In this paper, we repeat Goetz's experiment to measure the concentrations of glucose, urea, and lactic acid in aqueous solution using a confocal Raman microscopy system, and we evaluate the performance of this approach for multicomponent analysis.…”

Section: Confocal Raman Microscopymentioning

confidence: 99%

Quantifying the concentration of glucose, urea, and lactic acid in mixture by confocal Raman microscopy

Tang

Barton

Ward

et al. 2018

Biophotonics: Photonic Solutions for Better Health Care VI

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Raman spectroscopy has numerous applications in the field of biology. One such application is the simultaneously measurement of the concentration of multiple biochemical components in low volume aqueous mixtures, for example, a single drop of blood serum. Over twenty years ago, it was shown for the first time that it was possible to estimate the concentration of glucose, urea, and lactic acid in mixture by combining Raman Spectroscopy with Partial Least Squares Regression analysis. This was followed by numerous contributions in the literature designed to increase the number of components and reduce the limits of concentration that could be simultaneously measured using Raman spectroscopy, by developing various optical architectures to maximise the signal to noise ratio. The aim of this paper is to demonstrate the potential of a confocal Raman microscopy system for multicomponent analysis for the case of physiologically relevant mixtures of glucose, urea, and lactic acid.

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Confocal microscopy and exfoliative cytology

Cited by 7 publications

References 9 publications

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

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

Multicomponent analysis using a confocal Raman microscope

Quantifying the concentration of glucose, urea, and lactic acid in mixture by confocal Raman microscopy

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