Surface fractal computation and its application to immunofluorescent histochemical studies of calpain and calpastatin in PC12 cells

Surface fractal computation and its application to immunofluorescent histochemical studies of calpain and calpastatin in PC12 cells

The purpose of this report is to present a method which can be used to parameterize patterns of immunofluorescent staining in cultured neural cells. The algorithm is based on the observation that the variance in pixel intensity of the image is a power function of the magnitude of the area in immunof...

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Bibliographic Details
Published in:Journal of neuroscience methods Vol. 103; no. 2; pp. 191 - 197
Main Authors: DePetrillo, Paolo B, Yang, Qingfeng, Rackoff, Jamie, SanMiguel, Anitza, Karimullah, Kamran
Format: Journal Article
Language:English
Published: Amsterdam Elsevier B.V 30-11-2000
Elsevier Science
Subjects:
Actin
Actins - metabolism
Actins - ultrastructure
Alcohol
Algorithms
Animals
Biological and medical sciences
Calcium-Binding Proteins - metabolism
Calcium-Binding Proteins - ultrastructure
Calpain
Calpain - metabolism
Calpain - ultrastructure
Calpastatin
Ethanol - pharmacology
Ethyl
Fluorescent Antibody Technique - instrumentation
Fluorescent Antibody Technique - methods
Fractal
Fractals
Fundamental and applied biological sciences. Psychology
General aspects. Models. Methods
Image Processing, Computer-Assisted - instrumentation
Image Processing, Computer-Assisted - methods
Linear Models
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Models, Biological
PC12 Cells - cytology
PC12 Cells - drug effects
PC12 Cells - metabolism
Rats
Vertebrates: nervous system and sense organs
Calpain
Alcohol
Fractal
Calpastatin
Actin
Ethyl
Peptidases
Cell line
Neuron
Enzyme
Cysteine endopeptidases
Hydrolases
Technique
Immunofluorescence
In vitro
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Summary:The purpose of this report is to present a method which can be used to parameterize patterns of immunofluorescent staining in cultured neural cells. The algorithm is based on the observation that the variance in pixel intensity of the image is a power function of the magnitude of the area in immunofluorescently stained PC12 cells. This property is used to derive the fractal dimension ( D) of the region of interest (ROI), and corresponds to the complexity of the pixel intensity associated with the ROI, which is analogous to a fractal surface. We show that the measure is useful in characterizing immunofluorescent staining patterns, and apply this measure to study the effects of ethanol exposure on μ-calpain and calpastatin-associated immunoreactivity. Exposure of PC12 cells to ethanol (80 mM) × 48 h resulted in alterations in immunofluorescent signal (Control vs ethanol) associated with actin, calpastatin and μ-calpain: 2289 ± 166 vs 1709 ± 69, P < 0.01; 1681 ± 38 vs 2224 ± 95, P < 0.001; 1823 ± 39 vs 2841 ± 68, P < 0.0001 respectively, magnitudes being pixel intensity units on a scale of 0–4095. D-values for the three proteins in the same order were: 2.32 ± 0.01 vs 2.31 ± 0.03, NS; 2.31 ± 0.01 vs 2.32 ± 0.01, NS; 2.16 ± 0.03 vs 2.24 ± 0.02, P < 0.01, with a possible D-value range of 2–3.
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ISSN:0165-0270
1872-678X
DOI:10.1016/S0165-0270(00)00317-4