The study of cerebral astrocyte spatial organization using confocal laser microscopy
D Ye KORZHEVSKIY , O V KIRIK , Ye G SUKHORUKOVA , A V GILIAROV , K V SOLOVYOV , N A GRUDININA
Morphology ›› 2009, Vol. 135 ›› Issue (3) : 76 -79.
The study of cerebral astrocyte spatial organization using confocal laser microscopy
Three-dimensional reconstruction of cytoarchitectonic relations of the main components of the nervous tissue appears to be an important, yet an extremely laborious task of neuromorphology. The aims of the present study were: to develop the method permitting accurate visualization of astrocytes in rat brain slices thicker then 30 μm, to create an adequate algorithm for the study of the slides received using a confocal microscope and to perform the three-dimensional reconstructions of astrocytes from Z-series of confocal images. Different variants of slide preparation (various clearing reagents, fluorochromes, mounting media) were investigated, together with the operating modes of confocal microscope, methods of digital image processing etc. The methodical approaches are recommended that may be successfully applied for three-dimensional reconstruction and detection of spatial relations of astrocytes in mammalian brain.
brain / astrocytes / confocal laser microscopy / three-dimensional reconstruction
| [1] |
Коржевский Д.Э. и Гиляров А.В. Иммуноцитохимическое выявление тканевых антигенов после длительного хранения объектов в метилсалицилате. Морфология, 2008, т. 134, вып. 6, с. 76-78. |
| [2] |
Коржевский Д.Э., Григорьев И.П. и Отеллин В.А. Применение обезвоживающих фиксаторов, содержащих соли цинка, в нейрогистологических исследованиях. Морфология, 2006, т. 129, вып. 1, с. 85-87. |
| [3] |
Коржевский Д.Э. и Отеллин В.А. Иммуноцитохимическое выявление астроцитов в срезах головного мозга в сочетании с окраской по Нисслю. Морфология, 2004, т. 125, вып. 3, с. 100-102. |
| [4] |
Лилли Р. Патогистологическая техника и практическая гистохимия. М., Мир, 1969. |
| [5] |
Ромейс Б. Микроскопическая техника. М., Изд-во иностр. лит-ры, 1954. |
| [6] |
Феофанов А.В. Спектральная лазерная сканирующая конфокальная микроскопия в биологических исследованиях. Успехи биол. химии, 2007, т. 47, с. 371-410. |
| [7] |
Lindig T.M., Kumar V., Kikinis R. et al. Spiny versus stubby: 3D reconstruction of human myenteric (type I) neurons. Histochem. Cell Biol., 2009, v. 131, № 1, p. 1-12. |
| [8] |
Periasamy A., Skoglung P., Noakes C. and Keller R. An evaluation of two-photon excitation versus confocal and digital deconvolution fluorescence microscopy imaging in Xenopus morphogenesis. Microsc. Res. Tech., 1999, v. 47, p. 172-181. |
| [9] |
Stachs O., Knappe S., Zhivov A. et al. 3D-konfokale Laser-Scanning-Mikroskopie der kornealen epithelialen Nervenstruktur. Klin. Monatsbl. Augenheilkd., 2006, Bd. 223, H. 7, S. 583-588. |
| [10] |
Vuksic M., Del Turco D., Bas Orth C. et al. 3D-reconstruction and functional properties of GFP-positive and GFP-negative granule cells in the fascia dentata of the Thy1-GFP mouse. Hippocampus, 2008, v. 18, № 4, p. 364-375. |
Eco-Vector
/
| 〈 |
|
〉 |