Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Comprehensive Review of Golgi Staining Methods for Nervous Tissue

  • Kang, Hee Won (Department of Medicine, Korea University College of Medicine) ;
  • Kim, Ho Kyu (Department of Medicine, Korea University College of Medicine) ;
  • Moon, Bae Hun (Department of Medicine, Korea University College of Medicine) ;
  • Lee, Seo Jun (Department of Medicine, Korea University College of Medicine) ;
  • Lee, Se Jung (Department of Anatomy, Korea University College of Medicine) ;
  • Rhyu, Im Joo (Department of Anatomy, Korea University College of Medicine)
  • Received : 2017.06.22
  • Accepted : 2017.06.30
  • Published : 2017.06.30
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Abstract

Golgi staining has been modified and developed since Camillo Golgi introduced the black reaction in 1873. This study focuses on the commonly used Golgi staining methods and presents comprehensive data regarding three Golgi staining methods along with their strong and weak points. The Golgi-Cox method uses mercuric chloride for brain tissue impregnation and is a reliable technique for analyzing the complete dendritic tree of cortical neurons. However, specimens tend to shrink during the staining steps. Recent combination of the Golgi-Cox method and immunofluorescence provides additional options for neuroscientists. Rapid Golgi staining requires osmium tetroxide for the post-fixation process. It homogenously stains whole structures of neurons and provides their detailed anatomical morphology. This staining is influenced by the age of the specimen, temperature of the laboratory, and duration of each procedure. The Golgi-Kopsch method uses formaldehyde and glutaraldehyde instead of osmium tetroxide and can be used regardless of the age of the specimen and the duration after fixation. This method is suitable for research using human brain fixed for a long time or for specimens obtained from old-aged animals. Selecting a Golgi staining protocol that is appropriate for the specimen type and research purpose is important to achieve best results.

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References

  1. Ahn B K, Park M J, Kim E H, Bae Y C, Hong H S, Cho H J, and Joo K (1990) A study on staining techniques of Golgi methods. Kyungpook Univ. Med. J. 31, 313-322.
  2. Bentivoglio M, Jones E G, Mazzarello P, Ribak C E, Shepherd G M, and Swanson L W (2011) Camillo Golgi and modern neuroscience. Brain Res. Rev. 66, 1-4. https://doi.org/10.1016/j.brainresrev.2010.12.001
  3. Buell S J (1982) Golgi-Cox and rapid golgi methods as applied to autopsied human brain tissue: widely disparate results. J. Neuropathol. Exp. Neurol. 41, 500-507. https://doi.org/10.1097/00005072-198209000-00003
  4. De Carlos J A and Borrell J (2007) A historical reflection of the contributions of Cajal and Golgi to the foundations of neuroscience. Brain Res. Rev. 55, 8-16. https://doi.org/10.1016/j.brainresrev.2007.03.010
  5. Garin-Aguilar M E, Diaz-Cintra S, Quirarte G L, Aguilar-Vazquez A, Medina A C, and Prado-Alcala R A (2012) Extinction procedure induces pruning of dendritic spines in CA1 hippocampal field depending on strength of training in rats. Front. Behav. Neurosci. 6, 12.
  6. Glickstein M (2006) Golgi and Cajal: the neuron doctrine and the 100th anniversary of the 1906 Nobel Prize. Curr. Biol. 16, R147-R151. https://doi.org/10.1016/j.cub.2006.02.053
  7. Golgi C (1873) On the structure of the brain grey matter. Gazzetta Medica Italiana. Lombardia 33, 244-246.
  8. Jones E G (2011) Cajal's debt to Golgi. Brain Res. Rev. 66, 83-91. https://doi.org/10.1016/j.brainresrev.2010.04.005
  9. Koyama Y (2013) The unending fascination with the Golgi method. OA Anatomy 1, 8.
  10. Koyama Y and Tohyama M (2012) A modified and highly sensitive Golgi-Cox method to enable complete and stable impregnation of embryonic neurons. J. Neurosci. Methods 209, 58-61. https://doi.org/10.1016/j.jneumeth.2012.06.007
  11. Koyama Y and Tohyama M (2013) A novel, Golgi-Cox-based fluorescent staining method for visualizing full-length processes in primary rat neurons. Neurochem. Int. 63, 35-41. https://doi.org/10.1016/j.neuint.2013.04.009
  12. Maiti P, Manna J, and McDonald M P (2015) Merging advanced technologies with classical methods to uncover dendritic spine dynamics: A hot spot of synaptic plasticity. Neurosci. Res. 96, 1-13. https://doi.org/10.1016/j.neures.2015.02.007
  13. Narayanan S N, Jetti R, Gorantla V R, Kumar R S, Nayak S, and Bhat P G (2014) Appraisal of the effect of brain impregnation duration on neuronal staining and morphology in a modified Golgi-Cox method. J. Neurosci. Methods 235, 193-207. https://doi.org/10.1016/j.jneumeth.2014.07.007
  14. Oda S I, Lee KJ, Arii T, Imoto K, Hyun B H, Park I J, Kim H, and Rhyu I J (2010) Differential regulation of Purkinje cell dendritic spines in rolling mouse Nagoya (tgrol/tgrol), P/Q type calcium channel (${\alpha}1A$/ Cav2.1) mutant. Anat. Cell Biol. 43, 211-217. https://doi.org/10.5115/acb.2010.43.3.211
  15. Pannese E (1999) The Golgi Stain: invention, diffusion and impact on neurosciences. J. Hist. Neurosci. 8, 132-140. https://doi.org/10.1076/jhin.8.2.132.1847
  16. Pannese E (2007) The contribution of Camillo Golgi to our understanding of the structure of the nervous system. Arch. Ital. Biol. 145, 111-115.
  17. Patro N, Kumar K, and Patro I (2013) Quick Golgi method: modified for high clarity and better neuronal anatomy. Indian J. Exp. Biol. 51, 685-693.
  18. Ramon-Moliner E (1970) The Golgi-Cox technique. In: Contemporary Research Methods in Neuroanatomy, ed. Nauta W J H and Ebbesson S O E, pp. 32-55, (Springer-Verlag, New York).
  19. Rhyu Y S, Lee S J, Kim D H, and Uhm C S (2016) Application of threedimensional light microscopy for thick specimen studies. Appl. Microsc. 46, 93-99. https://doi.org/10.9729/AM.2016.46.2.93
  20. Riley J N (1979) A reliable Golgi-Kopsch modification. Brain Res. Bull. 4, 127-129. https://doi.org/10.1016/0361-9230(79)90067-4
  21. Rosoklija G, Mancevski B, Ilievski B, Perera T, Lisanby S H, Coplan J D, Duma A, Serafimova T, and Dwork AJ (2003) Optimization of Golgi methods for impregnation of brain tissue from humans and monkeys. J. Neurosci. Methods 131, 1-7. https://doi.org/10.1016/j.jneumeth.2003.06.001
  22. Scheibel M E and Scheibel A R (1978) The Methods of Golgi. In: Neuroanantomical Resarch Techniques, pp. 89-114, (Academic Press, New York).
  23. Smitha J and Roopa R (2012) Rapid Golgi Technique for staining pyramidal neurons in rat hippocampus. IJBAMS 2, 98-102.
  24. Spiga S, Acquas E, Puddu M C, Mulas G, Lintas A, and Diana M (2011) Simultaneous Golgi-Cox and immunofluorescence using confocal microscopy. Brain Struct. Funct. 216, 171-182. https://doi.org/10.1007/s00429-011-0312-2
  25. Tokuno H, Nakamura Y, Kudo M, and Kitao Y (1990) Effect of Triton X-100 in the Golgi-Kopsch method. J. Neurosci. Methods 35, 75-77. https://doi.org/10.1016/0165-0270(90)90096-X
  26. Valverde F (1970) The Golgi Method. A Tool for comparative Structural Analyses (Springer, New York).
  27. Zaqout S and Kaindl A M (2016) Golgi-Cox staining step by step. Front. Neuroanat. 10, 38.