Applications in cell research

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Wolf D. Kuhlmann
 
Institut für Immunbiologie, Freiburg
Institut für Nuklearmedizin DKFZ,
Heidelberg, Germany
 
The development of alpha-1-fetoprotein (AFP)-producing cells was studied in rat liver by ultrastructural immunoperoxidase cytochemistry. AFP was detected in fetal and postnatal hepatocytes, including cell stages which ranged from the fetal to the adult cell-type. Irrespective of the stage of hepatocyte maturation, AFP was localized in the perinuclear space, the rough-surfaced endoplasmic reticulum (RER), and the Golgi apparatus. Non-membrane-bound ribosomes did not stain for AFP. Between Days 3 and 8 after birth, hepatocytes became comparable to those seen in adult liver. After Day 8, more and more unstained hepatocytes were observed, but AFP-staining cells with mature hepatocyte characteristics were still present. On Day 21, AFP was seen in short segments of the RER in few hepatocytes. After Day 28, AFP-staining cells were no longer detected. During postnatal liver maturation, AFP production was not due to the generation of morphologically distinct fetal-type hepatocytes.
 

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Wolf D. Kuhlmann
 
Division of Radiooncology, Deutsches Krebsforschungszentrum,
69120 Heidelberg, Germany

 

Buffer solutions have the useful property of resisting changes in pH when hydrogen ions are added to or subtracted from the solution by chemical reactions or by other means. This is because a buffer solution contains both an acidic and a basic component. Buffer solutions are important because many reactions will work best within a cerain range of pH values. Aqueous buffer solutions can be prepared by several methods with the basic principle of all of them: the solution must contain the acid and the base of a conjugate in a reasonably similar and reasonably high concentration. In the direct method, the buffer is prepared by mixing the appropriate moles of “conjugate acid” with the appropriate number of moles of “conjugate base”. If the conjugate acid and base are not available, the buffer can be prepared by a indirect method, i.e. by partial conversion of the conjugate acid to conjugate base (and vice versa) using a strong acid or base as appropriate. In practice, biochemical buffers which cover a range of defined pH-values are prepared from stock solutions of the respective substances by mixing of appropriate amounts.

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Wolf D. Kuhlmann

Immunocytochemistry SFB 136 and Institut für Nuklearmedizin, DKFZ
Heidelberg, Germany
 
Hepatomas were induced in rats by feeding N-nitrosomorpholine. Nodules from alpha-1-fetoprotein (AFP) producing hepatoma were taken and subcutaneously injected into syngeneic hosts. Transplanted hepatomas of the 14th to 19th generations were used for ultrastructural localization of AFP. Various methods of tissue processing were examined under the light and electron microscopes for intracellular detection of AFP by use of specific anti-AFP antibodies conjugated with peroxidase. AFP was localized in the perinuclear space, the rough-surfaced endoplasmic reticulum and the Golgi apparatus.
 

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