Quantitative energy-filtered image analysis in cytochemistry. II. Morphometric analysis of element-distribution images.
A combination of energy-filtered electron microscopy (EFEM) and an image-analysis system (IBAS/2000) is used for a morphometric analysis of chemical reaction products in cells. Electron energy-loss spectroscopic element-distribution images are acquired from cytochemical reaction products in a variety of cellular objects: (1) colloidal thorium particles in extra-cellular coat material, (2) iron-containing ferritin particles in liver parenchymal cells, (3) barium-containing reaction products in endoplasmic reticulum stacks, (4) elements present in lysosomal cerium- and barium-containing precipitates connected with acid phosphatase (AcPase) or aryl sulphatase (AS) enzyme activity. Areas or area fractions are determined from such element-distribution images by application of an objective image segmentation method. By superposition of two or more element-distribution images, mutual element relations are qualitatively established in lysosomal cerium- and barium-containing precipitates connected with acid phosphatase (AcPase) or aryl sulphatase (AS) enzyme activity. By comparing electron spectroscopic images (ESI) with element-distribution images, the mutual contrast per element relations are quantitatively investigated. The obtained gain in resolution in such electron energy-loss spectroscopic element-distribution images will be explained and discussed.
Sorber
C W
CW
AEM Unit, Clinical Pathological Institut I. Erasmus University Rotterdam, The Netherlands.
Van Dort
J B
JB
Ringeling
P C
PC
Cleton-Soeteman
M I
MI
De Bruijn
W C
WC
D000818:Animals; D002478:Cells, Cultured; D004848:Epithelium / Q000648:ultrastructure; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008099:Liver / Q000648:ultrastructure*; D008264:Macrophages / Q000648:ultrastructure*; D051379:Mice; D008854:Microscopy, Electron; D051381:Rats; D014621:Vagina / Q000648:ultrastructure*