Abstract
The usefulness of computer-assisted image analysis with particular emphasis on fluorescence microscopy was evaluated and exemplified. Problems associated with image pick-up and transfer between microscope and computer are discussed. The importance of fully supported software programs adjusted to the needs of histology are emphasized. One such system, the IBAS (Kontron/Zeiss, West Germany) has been found suitable for use also by people with little or no background computer knowledge. Three examples where image analysis clearly adds a unique quantitative dimension to the evaluation of the results have been presented. (1) Nerve density measurements: A semi-automatic interactive program was used to evaluate the potentially neurotoxic effects of hexachlorophene and chlorhexidine, two disinfectant agents, using an intraocular screening model in which the density of the sympathetic autonomic ground plexus of the iris is studied by Falck-Hillarp fluorescence histochemistry applied to iris whole mounts. Pronounced neurotoxic effects were described. Dopaminergic nerve density measurements in striatum following neurotoxic drug treatments correlate well with other measurements of degree of denervation. (2) Transmitter release and diffusion: Experimentally induced unilateral parkinsonism in rats can be counteracted by intrastriatal implants of chromaffine tissue. These grafts work by releasing large quantities of catecholamines which diffuse through host neuropil. Image analysis was used to characterize in detail the diffusion of catecholamines using Falck-Hillarp fluorescence histochemistry. Linear scans of fluorescence intensity and imaging fluorescence gradients using false color look-up tables enables fast visual quantitative interpretation of the results. (3) Morphometry of smeared and sectioned astrocytes: A program was used that calculated area and perimeter of smeared astrocytes stained with an antiserum against glial fibrillary acidic protein, GFA. A study of astrocyte growth from adolescence to senescence revealed continuous growth of astrocytes throughout life. In this case the extreme complexity of astrocyte morphology necessitated special interactive procedures to be used in which the experimenter can “retouch” the digitized image prior to binary transformation. Area and perimeter data of this kind could not have been obtained without an image processing system. Astrocyte overgrowth in brain tissue grafts to the anterior chamber of the eye and to the brain were also described. It is concluded that image analysis is a powerful tool for the quantitative evaluation of microscopical images with general usefulness in flourescene histochemistry.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Björklund, A., Dunnett, S.B., Stenevi, U., Lewis, M.E. and Iversen, S.D. (1980). Reinnervation of the denervated striatum by substantia nigra transplants: functional consequences as revealed by pharmacological and sensorimotor testing. Brain Res. 199, 307–333.
Björklund, H. and Dahl, D. (1982). Glial disturbances in isolated neocortex: Evidence from immunohistochemistry of intraocular grafts Develop. Neurosci., 5, 424–435.
Björklund, H., Dahl, D., Haglid, K., Rosengren, L. and Olson, L. (1983a). Astrocytic development in fetal parietal cortex grafted to cerebral and cerebellar cortex of immature rats. Develop. Brain Res 9, 171–180.
Björklund, H., Dahl, D. and Olson, L. (1984a). Morphometry of GFA and vimentin positive astrocytes in grafted and lesioned cortex cerebri. Intern. J. Developm. Neurosci., 2, 181–192.
Björklund, H., Eriksdotter-Nilsson, M., Dahl, D. and Olson, L. (1984b). Astrocytes in semars of CNS tissues as visualized by GFA and vimentin immunofluorescence. Med. Biol., 62, 38–48.
Björklund, H., Eriksdotter-Nilsson, M., Dahl, D. and Olson, L. (1984b). Astrocytes in semars of CNS tissues as visualized by GFA and vimentin immunofluorescence. Med. Biol., 62, 38–48.
Björklund, H., Hoffer, B., Olson, L. and Seiger, A. (1981). Differential morphological changes in sympathetic nerve fibers elicited by lead, cadmium and mercury. Environmental Res., 26, 69–80.
Björklund, H., Seiger, A., Hoffer, B. and Olson, L. (1983b). Trophic effects of brain areas on the developing cerebral cortex. I. Growth and histological organization of intraocular grafts. Developm. Brain Res., 6, 131–140.
Corrodi, H. and Jonsson, G. (1967). The formaldehyde fluorescence method for the histochemical demonstration of biogenic monoamines. A review on the methodology. J. Histochem. Cytochem., 15 65–78.
Falck, B., Hillarp, N.-Å., Thieme, G. and Torp, A. (1962). Fluores¬cence of catecholamines and related compounds condensed with formaldehyde. J. Histochem. Cytochem., 10 348–354.
Freed, W., Ko, G., Neihoff, D., Kuhar, M., Hoffer, B., Olson, L., Spoor, E., Morihisa, J. and Wyatt, R. (1983). Normalization of spiroperidol binding in the denervated rat striatum by homologous substantia nigra transplants. Science, 222, 937–939.
Freed, W., Morihisa, J., Spoor, E., Hoffer, B., Olson, L., Seiger, Å. and Wyatt, R. (1981). Transplanted adrenal chromaffin cells in rat brain reduce lesion-induced rotational behaviour. Nature (Lond.), 292, 351–352.
Henschen, A. and Olson, L. (1983). Hexachlorophene-induced degeneration of adrenergic nerves: Application of quantitative image analysis to Falck-Hillarp fluorescence histochemistry. Acta Neuropathol. ( Berl. ), 59, 109–114.
Henschen, A. and Olson, L. (1984). Chlorhexidine-induced degeneration of adrenergic nerves. Acta Neuropathol. ( Berl. ), 63, 18–23.
Herrera-Marschitz, M., Stromberg, I., Olsson, D., Ungerstedt, U. and Olson, L. (1984). Adrenal medullary implants in the dopamine-denervated rat striatum. II. Acute behavior as a function of graft amount and location and its modulation by neuroleptics. Brain Res., 297 53–61.
Herrera-Marschitz, M., Stromberg, I., Olsson, D., Ungerstedt, U. and Olson, L. (1984). Adrenal medullary implants in the dopamine-denervated rat striatum. II. Acute behavior as a function of graft amount and location and its modulation by neuroleptics. Brain Res., 297 53–61.
Herrera-Marschitz, M., Stromberg, I., Olsson, D., Ungerstedt, U. and Olson, L. (1984). Adrenal medullary implants in the dopamine-denervated rat striatum. II. Acute behavior as a function of graft amount and location and its modulation by neuroleptics. Brain Res., 297 53–61.
Olson, L., Hamberger, B., Jonsson, G. and Malmfors, T. (1968). Combined fluorescence histochemistry and 3H -noradrenalin measurements of adrenergic nerves. Histochemie, 15, 38–45.
Olson, L. and Malmfors, T. (1970). Growth characteristics of adrenergic nerves in the adult rat. Fluorescence histochemical and 3H-noradrenaline uptake studies using tissue transplants to the anterior chamber of the eye. Acta Physiol. Scand., Suppl., 348, 1–112.
Palmer, M., Bjorklund, H., Olson, L. and Hoffer, B. (1983). Trophic effects of brain areas on the developing cerebral cortex. II. Elec- trophysiology of intraocular grafts. Developm. Brain Res., 6, 141–148.
Perlow, M., Freed, W., Hoffer, B., Seiger, A., Olson, L. and Wyatt, R. (1979). Brain grafts reduce motor abnormalities produced by de¬struction of nigrostriatal dopamine system. Science, 204 643–647.
Strömberg, I., Herrera-Marschitz, M., Hultgren, L., Ungerstedt, U. and Olson L. (1984). Adrenal medullary implants in the dopamine- denervated rat striatum. I. Acute catecholamine levels in grafts and host caudate as determined by HPLC-electrochemistry and fluorescence histochemical image analysis. Brain Res., 297, 41–51.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1985 The Wenner-Gren Centre
About this chapter
Cite this chapter
Olson, L., Björklund, H., Eriksdotter-Nilsson, M., Henschen, A., Strömberg, I. (1985). Image Analysis of Neuronal and Glial Markers: Fluorescence Microscopical Applications. In: Agnati, L.F., Fuxe, K. (eds) Quantitative Neuroanatomy in Transmitter Research. Wenner-Gren Center International Symposium Series, vol 42. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2139-2_17
Download citation
DOI: https://doi.org/10.1007/978-1-4613-2139-2_17
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4612-9263-0
Online ISBN: 978-1-4613-2139-2
eBook Packages: Springer Book Archive