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8 Tincion Simple

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La tinción simple es una tinción sencilla en la cual la bacteria es teñida con un sólo
reactivo. Es preferible utilizar tintes básicos que contengan un cromógeno (compuesto
que da color al tinte) con carga positiva, ya que la pared celular de las bacterias posee
componentes con carga negativa que atraen y enlazan el cromógeno. La tinción simple
se utiliza para observar la morfología y arreglo en el crecimiento de las bacterias.
Los tintes a utilizarse son:
1. Safranina
2. Cristal Violeta
3. Azul de Metileno

Cultivos en agar triptico de soya:
Escherichia coli
Bacillus subtilis
Staphylococcus aureus

1. Prepare un "frotis" del cultivo. (IMPORTANTE, este primer paso es uno que se va
a utilizar en la mayoría de las tinciones que se harán en el laboratorio.)
a. Si el medio es sólido:
i. Coloque una gota de agua con la aguja de inocular en el centro de la
ii. Transfiera una pequeña cantidad del cultivo a la gota de agua.
(utilice medidas asépticas y tenga mucho cuidado de no romper el
iii. Esparza sobre la laminilla.
iv. Espere que seque.
v. Mate y fije los organismos flameando.
b. Si el medio es líquido:
i. Siguiendo las medidas asépticas necesarias, coloque una gota de
cultivo en la laminilla.
ii. Esparza la gota sobre la laminilla.
iii. Espere que se seque.
iv. Flamee la laminilla para matar y fijar las bacterias.
2. Eche el tinte a utilizarse sobre el "frotis". Deje por un minuto.
3. Remueva e1 exceso de tinte con agua.
4. Seque cuidadosamente con bibulous paper.
5. Examine en el microscopio.

8 Tincion Simple



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