Fecha: 29 de agosto de 2023

Título: Uso y manejo de micropipetas y técnicas asépticas

Objetivos:

• Lograr una protección exitosa de un cultivo

• Transferir bacterias de un medio a otro fresco
• Lograr técnicas asépticas exitosas para proteger debidamente los cultivos

Notas/Anotaciones:

• Técnicas ascépticas:
o Logran descontaminar cierta superficie para que microorganismos
patogénicos no logren regenerarse.
o Inoculación
• Bacteria por utilizar: Micrococcus Luteus
• Transferencia
o Importante “spin” con vortex

Métodos:

I. Observations before each procedure:

Adjust flame to
Observe bacterial growth Follow proper Observe bacterial
Light Bunsen about 7cm (2 Get sample and
in original culture and asceptic growth in new
burner 3/4") with visible transfer properly
record appearance tecniques culture
central blue cone
II. Transferring to broth

Pick tubes •Loosen caps of bacterial culture and nutrient broth but do not remove
them

•Pick bacterial culture tube with nondominant hand and loop with
Sterilize dominant
•Hold loop in burner flame until it glows orange and do the same for wire

Get •Take lid off container and pass mouth through flame
•Dip loop into liquid culture
sample •Flame mouth again and close

• Take broth tube

• pass mouth through flame

Transfer
• Dip loop with bacteria into tube
• agitate with the loop to loosen bacterial cells and disperse them into the broth.
• Withdraw the loop from the broth and tap it against the inside of the tube to dislodge any
excess broth.
• Withdraw the loop from the tube.

Sterilize •Pass mouth through flame

•pass loop through flame to get rid of bacteria left on loop.

III. Transferring to an agar slant

Pick •Loosen caps of bacterial culture and nutrient agar but do not remove them
tubes
•Hold loop in burner flame until it glows orange and do the same for wire
•Place the bacterial culture tube and the nutrient agar slant tube in the palm of your
nondominant hand with the bottoms of the tubes against your palm.
Sterilize •The bacterial culture tube should nestle between your index and middle fingers and
the nutrient agar slant tube between your middle and little fingers.
•Use pressure from your thumb to hold the tubes in place.

•Pass the mouths of both tubes through the burner flame.

Get
•Insert the loop into the bacterial culture tube and pick up a small sample of culture
sample on the loop. Withdraw the loop from the tube.

•Insert the loop into the agar slant tube and, beginning at the lower end of the slant, drag the
loop across the agar in a zigzag pattern as you work toward the upper end of the slant. This
inoculates as much of the agar’s surface with bacteria as possible. Try not to dig into the agar
Transfer with the loop as you do so
•Flame the mouths of the tubes and replace the caps.
•Flame the loop and return it to the beaker.
•Return both tubes to the rack.

•Flame the mouths of the tubes and replace the caps.

Sterilize •Flame the loop and return it to the beaker.
•Return both tubes to the rack.
 Transfering to a plate

Pick •Loosen but do not remove the cap on the bacterial culture.
•Pick up the bacterial culture tube with your nondominant hand and the loop with your dominant
tubes hand.

•Flame the loop.

•Grasp the cap of the bacterial culture tube between the little and ring fingers of your dominant
Sterilize hand and remove the cap. Continue to hold the cap as you proceed.
•Pass the mouth of the bacterial culture tube through the burner flame.

•Insert the loop into the bacterial culture tube and pick up a small sample of culture on the loop.
Get •Withdraw the loop from the tube.
sample •Flame the mouth of the tube and replace the cap.
•Return the bacterial culture tube to the rack.

•Lift the lid of the petri dish just enough to insert the loop. Never completely uncover the dish. It
is best to open the dish clam-shell style. Drag the loop over the agar in a zigzag pattern to cover
Transfer as much of the agar’s surface as possible. Try not to dig into the agar with the loop as you do so.
•Withdraw the loop and replace the lid.

Sterilize •Flame the loop.

Observations after each procedure:

Return all equipment to

Record here the
Take your new cultures to the area
temperature at which they
indicated by your lab instructor for
will be incubated and any
incubation.
other details.
Return your starter culture
to where you picked it up
or dispose of it as
instructed.
proper storage. Wipe
down your workbench
with disinfectant. Wash
and dry your hands before
leaving the lab.
Observaciones:

Transferencia a caldo
• Se observó que en el medio fresco se formó un
precipitado en el tubo.

Transferencia a plato
• Se observaron resultados notables en el
crecimiento y la proliferación de estos
microorganismos.
• Se notó un incremento significa?vo en la
densidad de bacterias en comparación con el
estado inicial.

Conclusiones:
Luego de esta experiencia de laboratorio se determine que las técnicas de transferencia
fueron exitosas debido a que, al transferir las muestras al nuevo medio de cul?vo, Se observó la
formación de un precipitado en el caldo indicando entonces que el girar el medio original de las
bacterias y la técnica u?lizada para transferirlas fue exitosa. Es decir, se lograron transferir de un
medio a otro. Por otro lado, también se observó un crecimiento exitoso de bacterias en el plato
Petri. Además, se observe un crecimiento uniforme a través de la placa. Esto no tan solo indica la
eficiencia de las técnicas asép?cas u?lizadas para no contaminar la muestra, sino que además
demuestra que la transferencia al plato se llevó a cabo al pie de la letra.