Quantitative aspects of digital microscopy applied to cellular localization of heparin in smooth muscle cells

Richard F. Johnston; David K. Hanzel; Bob Stack; Brian Brandley; John Castellot

doi:10.1117/12.206818

12 April 1995 Quantitative aspects of digital microscopy applied to cellular localization of heparin in smooth muscle cells

Richard F. Johnston, David K. Hanzel, Bob Stack, Brian Brandley, John Castellot

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Proceedings Volume 2387, Advances in Laser and Light Spectroscopy to Diagnose Cancer and Other Diseases II; (1995) https://doi.org/10.1117/12.206818
Event: Photonics West '95, 1995, San Jose, CA, United States

Abstract

High Resolution digital acquisition allows a great deal of flexibility in the types of questions that can be directed to microscopic samples. To eliminate subjective bias and provide quantitative results we have approached microscopy with an automated digital format. This mode can return quantitative data at high resolution over large fields. The digital format makes accessible data including [data segmentation]: multispectral colocalization, seeding and connectivity, particle size and shape distribution and population analysis. We have begun a program to investigate this approach using the confocal microscope. Scanning larger fields-of-view at lower spatial resolutions (e.g., low magnification objective) defines large maps that allow alignment of high spatial resolution (diffraction limited) sampling. The [objective] selection of the field-of-view with low spatial resolution reduces the subjective nature of the selection of a 'typical staining pattern'. High resolution digital scanning in three dimensions contribute both to the 'objective' nature of the analysis and allow for quantitation of characteristics not historically available/accessible. The complex carbohydrate heparin is implicated in tumor growth and wound healing by affecting angiogenesis, cell proliferation and motility. The internal localization of heparin within vascular cells appears to be a good predictor of the sensitivity of those cells to the action of heparin. Cells resistant to the antiproliferative action of heparin are able to sequester the heparin in large vacuoles whereas those cells sensitive to the carbohydrate do not exhibit these structures. We have applied our approach to QUANTITATIVE DIGITAL MICROSCOPY to the analysis of intracellular heparin distribution.

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Richard F. Johnston, David K. Hanzel, Bob Stack, Brian Brandley, and John Castellot "Quantitative aspects of digital microscopy applied to cellular localization of heparin in smooth muscle cells", Proc. SPIE 2387, Advances in Laser and Light Spectroscopy to Diagnose Cancer and Other Diseases II, (12 April 1995); https://doi.org/10.1117/12.206818

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KEYWORDS

Microscopy

Spatial resolution

Microscopes

Particles

Statistical analysis

3D acquisition

3D scanning

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