Skip to main content
SpringerLink
Log in

An indirect method for quantitation of cellular zinc content of timm-stained cerebellar samples by energy dispersive X-ray microanalysis

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

The absolute concentration of zinc in the Purkinje cells of the rat cerebellum was determined by means of energy dispersive X-ray microanalysis (EDAX). Gelatine blocks with known zinc concentrations were stained by Timm's sulphide-silver method, and their silver concentrations were measured by EDAX. A linear correlation was found between the zinc and silver concentrations and this linear function was used as a quantitative calibration for evaluation of sulphide-silver staining, after perfusion with sodium-sulphide solution, fixation with glutaraldehyde, cryostat sectioning and staining of cerebellar samples in Timm's reagent.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Assaf SY, Chung S (1984) Release of endogenous Zn2+ from brain tissue during activity. Nature 308:734–735

    Google Scholar 

  • Chan AWK, Minski MJ, Lai JCK (1983) An application of neutron activation analysis to small biological samples: simultaneous determination of thirty elements in rat brain regions. J Neurosci Methods 7:317–328

    Google Scholar 

  • Crawford IL, Connor JD (1973) Glutamate in the cat hippocampus: distribution and release. Fed Proc 32:696

    Google Scholar 

  • Cunnane SC, Horrobin DF (1980) Parenteral linoleic and γ-linoleic acids ameliorate the gross effects of zinc deficiency. Proc Soc Exp Biol Med 164:583–588

    Google Scholar 

  • Danscher G (1981) Histochemical demonstration of heavy metals. A revised version of the sulphide silver method suitable for both light and electronmicroscopy. Histochemistry 71:1–16

    Google Scholar 

  • Danscher G (1984) Do the Timm sulphide silver method and selenium method demonstrate zinc in the brain? In: Frederickson CJ, Howell GA, Kasarskis EJ (eds) The neurobiology of zinc. Part A: Physiochemistry, anatomy and techniques. Alan R Liss, New York, pp 273–287

    Google Scholar 

  • Danscher G, Haug F-MS, Fredens K (1973) Effect of diethyldithiocarbamate (DEDTC) on sulphide silver stained boutons. Reversible blocking of Timm's sulphide silver stain for “heavy” metals in DEDTC treated rats (light microscopy). Exp Brain Res 16:521–532

    Google Scholar 

  • Danscher G, Fjerdingstad EJ, Fjerdingstad E, Fredens K (1976) Heavy metal content in subdivisions of the rat hippocampus (zinc, lead and copper). Brain Res 112:442–446

    Google Scholar 

  • DeBoer T, Bruinvels J, Bouta IL (1979) Differential effects of GABA analogues and zinc on glutamate decarboxylase, 4-aminobutyric-2-oxoglutaric acid transaminase and succinate semialdehyde dehydrogenase in rat brain tissue. J Neurochem 33:597–601

    Google Scholar 

  • Donaldson J, St. Pierre T, Minnich JL, Barbeau A (1973) Determination of Na+, K+, Mg2+, Cu2+, Zn2+ and Mn2+ in rat brain region. Can J Biochem 51:87–92

    Google Scholar 

  • Engelbart K, Kief H (1970) Über das funktionelle Verhalten von Zink und Insulin in den B-Zellen des Rattenpankreas. Virchows Arch [B] 4:294

    Google Scholar 

  • Frederickson CJ, Manton WI, Frederickson MH, Howell GA, Mallory MA (1982) Stable-isotope dilution measurement of zinc and lead in rat hippocampus and spinal cord. Brain Res 246:338–341

    Google Scholar 

  • Frederickson CJ, Klitenick MA, Manton WI, Kirkpatrick JB (1983) Cytoarchitectonic distribution of zinc in the hippocampus of man and the rat. Brain Res 273:335–339

    Google Scholar 

  • Gurusinghe CJ, Ehrlich D (1986) Gelatine embedding of central nervous system tissue improves the quality of vibratome sections. Stain Technol 61:324–326

    Google Scholar 

  • Hall TA, Clarke Anderson H, Appleton T (1973) The use of thin specimens for X-ray microanalysis in biology. J Microsc (London) 99:177–181

    Google Scholar 

  • Haug F-MS (1967) Electron microscopical localization of the zinc in hippocampal mossy fibre synapses by a modified sulphidesilver procedure. Histochemie 8:355–368

    Google Scholar 

  • Henke BL (1959) Microstructure, mass and chemical analysis with 8 to 44 Angstrom X radiation. Adv X-ray Analysis 2:117

    Google Scholar 

  • Hu KH, Friede RL (1968) Topographic determination of zinc in human brain after by atomic absorption spectrophotometry. J Neurochem 15:677–685

    Google Scholar 

  • Ibata Y, Otsuka N (1969) Electron microscopical demonstration of zinc in the hippocampal formation using Timm's sulphide silver technique. J Histochem Cytochem 17:171–175

    Google Scholar 

  • Kemp K, Danscher G (1979) Multi-element analysis of the rat hippocampus by proton induced X-ray emission spectroscopy (phosphorus, sulphur, chlorine, potassium, calcium, iron, zinc, copper, lead, bromine and rubidium). Histochemistry 59:167–176

    Google Scholar 

  • Klitenick MA, Frederickson CJ, Manton WI (1983) Acid-vapor decomposition for determination of zinc in brain tissue by isotope dilution mass spectrometry. Anal Chem 55:921–923

    Google Scholar 

  • Kirchgessner M, Roth HP, Weigand E (1976) Biochemical changes in zinc deficiency. In: Prasad AS (ed) Trace elements in human health and disease, vol I, zinc and copper. Academic Press, New York, pp 189–227

    Google Scholar 

  • Kozma M, Ferke A, Kása P (1978) Ultrastructural identification of neural elements containing trace metals. Acta Histochem 62:142–154

    Google Scholar 

  • Kozma M, Szerdahelyi P, Kása P (1981) Histochemical detection of zinc and copper in various neurons of the central nervous system. Acta Histochem 69:12–17

    Google Scholar 

  • Lezzi M (1979) Elemental analysis of solid microscopic samples in the flameless atomic absorption cuvette. Histochemistry 62:337–345

    Google Scholar 

  • Topez-Garcia C, Molowny A, Pérez-Clausell J (1983) Volumetric and densitometric study in the cerebral cortex and septum of a lizard (Lacerta galloti) using the Timm method. Neurosci Lett 40:13–18

    Google Scholar 

  • The Merck Index (1976) An encyclopedia of chemicals and drugs. In: Windholtz M (ed) 9. edn. Merck, Rahway, NJ, USA, pp 564

    Google Scholar 

  • Pérez-Clausell J, Danscher G (1985) Intravesicular localization of zinc in rat telencephalic boutons. A histochemical study. Brain Res 337:91–98

    Google Scholar 

  • Prento P (1979) Metals and phosphates in chloragosomes of lumbicus terrestris and their possible physiological significance. Cell Tissue Res 196:123–134

    Google Scholar 

  • Riordan JF, Vallce BL (1976) Structure and function of zinc metalloenzymes. In: Prasad AS (ed) Trace elements in human health and disease, vol I, Zinc and copper. Academic Press, New York, pp 227–257

    Google Scholar 

  • Schmidt PF, Fromme HG, Pfefferkorn G (1980) LAMMA-investigation of biological and medical specimens. Scanning Electron Microscopy Part II. SEM Inc., AMF O'Hare, Chicago, IL, USA, pp 623–634

    Google Scholar 

  • Stengaard-Pedersen K, Fredens K, Larsson LI (1983) Comparative localization of enkephalin and cholecystokinin immunoreactivities and heavy metals in the hippocampus. Brain Res 273:81–96

    Google Scholar 

  • Stengaard-Pedersen K, Larsson LI, Fredens K, Rehfeld JF (1984) Modulation of cholecystokinin concentrations in the rat hippocampus by chelation of heavy metals. Proc Natl Acad Sci USA 81:5876–5880

    Google Scholar 

  • Storm-Mathisen J, Leknes AK, Bore AT, Vaaland JL, Edminson P, Haug F-MS, Ottersen OP (1983) First visualization of glutamate and GABA in neurones by immunocytochemistry. Nature 301:517–520

    Google Scholar 

  • Szerdahelyi P (1982) Lack of correlation between trace metal staining and trace metal content of the rat hippocampus following colchicine microinjection. Histochemistry 74:563

    Google Scholar 

  • Szerdahelyi P, Kása P (1983) Variations in trace metal levels in rat hippocampus during ontogenic development. Anat Embryol 167:141–149

    Google Scholar 

  • Szerdahelyi P, Kása P (1984) Histochemistry of zinc and copper. In: Bourne GH, Danielli JF (eds) International review of cytology, vol 89. Academic Press, New York, pp 1–33

    Google Scholar 

  • Szerdahelyi P, Kása P (1985) Demonstration of reduced levels of zinc in rat brain after treatment with d-amphetamine, but not after treatment with reserpine. Histochemistry 83:181–187

    Google Scholar 

  • Szerdahelyi P, Kása P (1986) Histochemical demonstration of copper in normal rat brain and spinal cord. Histochemistry 85:341–347

    Google Scholar 

  • Szerdahelyi P, Kása P (1987) Regional differences in the uptake of exagenous copper into rat brain after acute treatment with sodium diethyldithiocarbamate. A histochemical and atomic absorption spectrophotometric study. Histochemistry 86:627–632

    Google Scholar 

  • Timm F (1958) Zur Histochemie der Schwermetalle. Das Sulfid-Silber Verfahren. Dtsch Z Gesamte Gerichtl Med 46:706–711

    Google Scholar 

  • Torjussen W, Haug F-MS, Olsen A, Andersen I (1978) Topochemistry of trace metals in nasal mucosa. Potentialities of some histochemical methods and energy dispersive X-ray microanalysis. Acta Histochem 63:11–25

    Google Scholar 

  • Wensink J, Lenglet WJM, Vis RD, Van den Hamer CJA (1987) The effect of dietary zinc deficiency on the mossy fiber zinc content of the rat hippocampus. A microbeam PIXE study. Histochemistry 87:65–69

    Google Scholar 

  • Zs. Nagy I, Pieri C, Giuli C, Bertoni-Freddari C, Zs. Nagy V (1977) Energy dispersive X-ray microanalysis of the electrolytes in biological bulk specimen. J Ultrastruct Res 58:22–33

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Central Research Laboratory, Medical University, Somogyi B. u. 4, H-6720, Szeged, Hungary

    I. Farkas, P. Szerdahelyi & P. Kása

Authors
  1. I. Farkas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Szerdahelyi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Kása
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Farkas, I., Szerdahelyi, P. & Kása, P. An indirect method for quantitation of cellular zinc content of timm-stained cerebellar samples by energy dispersive X-ray microanalysis. Histochemistry 89, 493–497 (1988). https://doi.org/10.1007/BF00492607

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00492607

Keywords

Search

Navigation