Document Detail


Fully automated and fast image analysis of autoradiographs with a TAS-Leitz. Determination of size, Feulgen fluorescence and grain counts of individual nuclei and their evaluation by a simplified cluster analysis.
MedLine Citation:
PMID:  6500994     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A fully automatic analysis system based on television image analysis was developed to measure simultaneously three parameters in individual nuclei of microscopic autoradiographs prepared from mouse jejunal crypt cell squashes and ascites tumor cell smears: size, Feulgen fluorescence and reflection from silver grains. A dark light camera with an image intensified silicon tube (RCA-ISIT), an automatic scanning stage and an autofocus device were fitted to a Leitz-TAS microscope. The camera permitted localization of Feulgen stained nuclei and measurement of area and light intensity by means of incident of light fluorescence in the red. After automatic changes of the Opak-illuminator silver grains were determined by means of polarized incident light reflected from the grains in the blue. A 25 X oil objective (aperture 0.75) yielded sufficient resolution for measurements. The nadir between the proportions of labeled and unlabeled nuclei was calculated from the data of one specimen on a PDP-computer using a new algorithm based on the minimal variance of the logarithm of reflected light per nucleus. Labeling indices determined by visual grain counting and by automatic analysis of the autoradiographs were well correlated (r = 0.87 to 0.92). Visual grain counts/nucleus and reflected light/nucleus correlated well when individual nuclei were compared (r = 0.92 to 0.97) or means of labeled nuclei of various specimens prepared during a 5 year period (r = 0.90 to 0.93). Quenching of nuclear Feulgen fluorescence was minimal. The optimal labeling range is 30-100 grain counts/nucleus. The time interval between measurements of two specimens was 25 min for a squash of approximately 350 crypt cells within a 3 mm X 3 mm field, and 20 min for a meandering scan with 1,000 ascites tumor cells.
Authors:
E D Wachsmuth; M Becker; A P Grieve; B Maurer-Schultze
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Histochemistry     Volume:  81     ISSN:  0301-5564     ISO Abbreviation:  Histochemistry     Publication Date:  1984  
Date Detail:
Created Date:  1985-01-09     Completed Date:  1985-01-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0411300     Medline TA:  Histochemistry     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  227-36     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Autoradiography / instrumentation*,  methods
Cell Nucleus / metabolism*,  ultrastructure
Coloring Agents
Jejunum / metabolism
Leukemia L1210 / metabolism
Mice
Rosaniline Dyes*
Thymidine / metabolism
Tritium
Chemical
Reg. No./Substance:
0/Coloring Agents; 0/Feulgen stain; 0/Rosaniline Dyes; 10028-17-8/Tritium; 50-89-5/Thymidine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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