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BACKSCATTERED ELECTRON IMAGING OF CRYSTAL MATRIX PROTEIN ON THE SURFACE OF CALCIUM OXALATE CRYSTALS USING COLLOIDAL GOLD
Author(s) -
Suzuki Koji,
Nakajima Chiaki,
Moriyama Manabu,
Miyazawa Katsuhito,
Tsugawa Ryuzo
Publication year - 1995
Publication title -
international journal of urology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 67
eISSN - 1442-2042
pISSN - 0919-8172
DOI - 10.1111/j.1442-2042.1995.tb00430.x
Subject(s) - bovine serum albumin , calcium oxalate , scanning electron microscope , oxalate , immunogold labelling , crystal (programming language) , chromatography , oxalic acid , nuclear chemistry , materials science , electron microscope , chemistry , biochemistry , inorganic chemistry , optics , computer science , composite material , programming language , physics
In order to clarify the presence and localization of crystal matrix protein (CMP) upon calcium oxalate crystals, scanning electron microscopy (SEM) and backscattered electron imaging (BEI) techniques were used. This protein exhibits a remarkable affinity with calcium oxalate crystals and may be important in stone pathogenesis. In this paper, rabbit anti‐human CMP polyclonal antibody was used as first antibody, and for the second antibody, goat anti‐rabbit IgG conjugated with 20 nm immunogold was used. Freshly prepared crystals from male urine were fixed in SEM fixative, then blocked and washed with phosphate‐buffered saline and bovine serum albumin (PBS/BSA). First and second antibodies were reacted in PBS/BSA. Crystals were then dehydrated and finally coated for SEM study. The SEM technique showed bipyramidal shaped dihydrate calcium oxalate crystals in every sample and even at high magnification, colloidal gold could barely be seen. BE 1 clearly demonstrated the presence and localization of the gold on the surface of the crystals as well as on the macromol‐ecules eluted from the crystals by dissolving them in ethylenediamminetetraacetic acid solution.