Microscopia, mesoscopia e macroscopia em neuroanatomia · microscopia, mesoscopia e macroscopia em...
Transcript of Microscopia, mesoscopia e macroscopia em neuroanatomia · microscopia, mesoscopia e macroscopia em...
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MICROSCOPIA, MESOSCOPIA
E MACROSCOPIA EM
NEUROANATOMIA
PROF. DR. MARCUS ALEXANDRE MENDES LUZ ANATOMIA HUMANA E ANATOMIA PATOLÓGICA
FACULDADE DE MEDICINA - UNILAGO
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• Barnard – Robert – Brown
(Rodrigues, H., 2005)
Le Masurier
Formol 10%
Solução de Mulligan (Sulfato de cobre)
Soluçâo de Cloreto férrico 1%
Solução de Ferrocianeto de potássio 1%
Preservação em Solução ácida de formol 10%
Resultados: Substância branca, branco; substância cinzenta, azul turquesa.
*Mulligan JH. A method of staining for macroscopic study. J Anat London, 65:472-476, 1931.
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BigBrain: An Ultrahigh-Resolution 3D Human Brain Model
Katrin Amunts, Claude Lepage, Louis Borgeat, Hartmut Mohlberg, Timo Dickscheid,
Marc-Étienne Rousseau, Sebastian Bludau, Pierre-Louis Bazin, Lindsay B. Lewis, Ana-
Maria Oros-Peusquens, Nadim J. Shah, Thomas Lippert, Karl Zilles, Alan C. Evans.
Science, v. 340, pp. 1472-1475, 2013
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HISTOQUÍMICA CLÁSSICA
• Cajal: Neurônios, astrócitos, nervos.
• Golgi (Impregnação cromo-argêntea): neurônios-Purkinje, astrócitos.
• Hortega: astrócitos, micróglia, oligodendrócitos.
• Galocianina, Tionina, Nissl (Violeta Cresil): Substância de Nissl
• Holtzer: Fibras da glia, glia
• Hematoxilina Férrica Lopez: Mielina
• Weigert-Pal: Mielina
• LFB: Mielina
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• Luxol-fast-blue - Nissl (Klüver-Barrera) para mielina e neurônios.
(http://anatpat.unicamp.br/tecnicashistologicas.html#t27a)
Luxol-fast-blue 0,1%
Luxol-fast-blue MBS 0,1 g.
Etanol 95% 100 ml.
Dissolver o corante no álcool. Acrescentar 0,5 ml de ácido acético 10% por 100 ml de
solução.
Cresil – violeta 0,1%
Cresil violeta 0,1 g.
Água destilada 100 ml.
Imediatamente antes do uso, adicionar 15 gotas de ácido acético a 10%. Filtrar antes
do uso.
Carbonato de lítio 0,05 %
Carbonato de lítio 0,05 g.
Água destilada 100 ml.
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Procedimento:
1. Corar pela solução de Luxol fast blue durante a noite em estufa a 56%.
Etanol 95 %.
2. Lavar em água destilada.
3. Diferenciar com carbonato de lítio, cerca de 30 segundos.
Continuar a diferenciação em álcool 70% e ir agitando gentilmente até que a diferença
entre substância branca e cinzenta fique nítida.
Monitorar a mudança cromática à microscopia
4. Corar por solução de cresil violeta 6 min.
5. Desidratar rapidamente.
6. Montar
Resultados : mielina – azul turquesa; neurônios – azul violeta.
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• Nissl (Violeta Cresil)
( http://www.neurofisiologia.unifesp.br/protocolonissl.htm)
1. Xilol I - H2O destilada: 2 minutos
2. Corante cresil violeta: 2 a 7 minutos (controlar o tempo pela cor)
3. H2O destilada : 5 mergulhos
4. Álcool 50º: 5 mergulhos
5. Álcool 70º: 5 mergulhos
6. Diferenciador (Ac. Acético 1% etanol) – 1 mergulho
7. Álcool 70º : 1 minuto até Xilol II
8. Montar
OBS: Hematoxilina de Harris por 5 minutos, lavar bem em água corrente até sair o
excesso de corante, diferenciar em álcool e ácido (250 mL de álcool 70% + 2,5 mL
de acido clorídrico), passar rápido nesse diferenciador. Lavar bem em água corrente e
passar para eosina alcoólica por 3 minutos. Desidratar.
Resultados: Neurônios, violeta.
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REFERÊNCIAS BIBLIOGRÁFICAS
• https://bigbrain.loris.ca/images/media/BBvideo_HBM.mp4
• http://www.livescience.com/42227-3d-images-human-brain.html
• http://anatpat.unicamp.br/neuro1.html
• http://anatpat.unicamp.br/neuroanatindex.html
• http://www.thehumanbrain.info/brain/bn_brain_atlas/brain.html