Characterization of aeroallergen of Texas panhandle using scanning and fluorescence microscopy

Nabarun Ghosh; Mandy Whiteside; Chris Ridner; Yasemin Celik; C. Saadeh M.D.; Jeff Bennert

doi:10.1117/12.853145

7 June 2010 Characterization of aeroallergen of Texas panhandle using scanning and fluorescence microscopy

Nabarun Ghosh, Mandy Whiteside, Chris Ridner, Yasemin Celik, C. Saadeh M.D., Jeff Bennert

Proceedings Volume 7729, Scanning Microscopy 2010; 77291X (2010) https://doi.org/10.1117/12.853145
Event: Scanning Microscopy 2010, 2010, Monterey, California, United States

Abstract

Aeroallergens cause serious allergic and asthmatic reactions. Characterizing the aeroallergen provides information regarding the onset, duration, and severity of the pollen season that clinicians use to guide allergen selection for skin testing and treatment. Fluorescence Microscopy has useful approaches to understand the structure and function of the microscopic objects. Prepared slides from the pollen were observed under an Olympus BX40 microscope equipped with FITC and TRITC fluorescent filters, a mercury lamp source, an Olympus DP-70 digital camera connected to the computer with Image Pro 6.0 software. Aeroallergens were viewed, recorded and analyzed with DP Manager using the Image Pro 6.0 software. Photographs were taken at bright field, the fluorescein-isothiocyanate (FITC) filter, and the tetramethylrhodamine (TRITC) filter settings at 40X. A high pressure mercury lamp or UV source was used to excite the storage molecules or proteins which exhibited autofluorescence. The FITC filter reveals the green fluorescent proteins (GFP and EGFP), and the TRITC filter for red fluorescent proteins (DsRed). SEM proved to be useful for observing ultra-structural details like pores, colpi, sulci and ornamentations on the pollen surface. Samples were examined with an SEM (TM-1000) after gold coating and Critical Point Drying. Pollen grains were measured using the TM-1000 imaging software that revealed the specific information on the size of colpi or sulci and the distance between the micro-structures. This information can be used for classification and circumscription in Angiosperm taxonomy. Data were correlated clinical studies established at Allergy A.R.T.S. Clinical Research Laboratory.

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Nabarun Ghosh, Mandy Whiteside, Chris Ridner, Yasemin Celik, C. Saadeh M.D., and Jeff Bennert "Characterization of aeroallergen of Texas panhandle using scanning and fluorescence microscopy", Proc. SPIE 7729, Scanning Microscopy 2010, 77291X (7 June 2010); https://doi.org/10.1117/12.853145

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KEYWORDS

Scanning electron microscopy

Luminescence

Meteorology

Microscopy

Microscopes

Optical filters

Carbon dioxide

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