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Histochemistry

, Volume 42, Issue 1, pp 9–29 | Cite as

High-resolution scanning-densitometry of photographic negatives of human metaphase chromosomes

I. Instrumentation
  • M. van der Ploeg
  • P. van Duijn
  • J. S. Ploem
Article

Summary

A computer-controlled cytophotometrical system is described to determine local absorbance or fluorescence values of chromosomes.

The absorbance or fluorescence values within the chromosome are obtained in digital form by interactive scanning of 35 mm photomicrographic negatives. The scanning system consists of a mechanical stage scanner (Scanning Microscope Photometer SMP, Zeiss) connected to a PDP-12 computer (Digital Equipment).

A computer program (CHROSCAN) enables the scanning of single chromosome images in a direction perpendicular to the length axis with 0.16 μ increments at the specimen level, and a transmission resolution of 500 linearly spaced intensity levels. Optical density values corrected for the local background of the individual chromosome, and total integrated optical density as well as the ratio of optical density in the long arm over the total optical density are calculated. Photography of a grey wedge together with the metaphase, allows determination of the gamma of each individual photomicrograph and conversion of optical density values into actual absorbance values.

The program has a papertape output of the corrected integrated optical density data per lateral scanline to be used as an input for a curve analysis program. It also allows plotting of the integrated optical density distribution over the chromosome length by a Calcomp plotter.

In this paper the instrumentation and experimental set up are presented, together with data concerning the reproducibility and accuracy of the photographic and the scanning procedures.

Keywords

Chromosome Length Integrate Optical Density Digital Equipment Optical Density Data Total Optical Density 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1974

Authors and Affiliations

  • M. van der Ploeg
    • 1
  • P. van Duijn
    • 1
  • J. S. Ploem
    • 1
  1. 1.Department of Histochemistry and CytochemistryUniversity of LeidenLeidenThe Netherlands

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