Unit 1.1f Flashcards
What is the point in aseptic techniques?
Prevent contamination by microorganisms of cell cultures and also prevent accidental inoculation
What are microbe’s cultured on?
Broth (suspended liquid medium) or on /within agar (solid medium)
What do different media’s promote?
The growth of different cell types and microbes
What are optimum growth conditions provided in?
Nutrients, pH, gasses
What is step 1 in inoculation?
Wash hands with antibacterial soap or alcohol to reduce bacteria
What is step 2 in inoculation?
Swap area you are working in with disinfectant to sterilise the area
What is step 3 in inoculation?
Using permanent pen label base of agar plate:
your name, microbes name, date, class
What is step 4 in inoculation?
Flame metal inoculating hoop until it glows red
What is step 5 in inoculation?
Remove lid off the culture vessel using pinky finger, gently flame lip of culture vessel
What is step 6 in inoculation?
In the Bunsens air canopy, pick up the sterile hoop and remove some of the yeast culture
What is step 7 in inoculation?
Keep hold of loop, reframe lip and replace lid
What is step 8 in inoculation?
Working in the Bunsen’s canopy, pick up base of sterile agar plate and with inoculating loop gently streak a pattern to dilute your culture of yeast microbes
What is step 9 in inoculation?
Place face of inoculated agar plate onto its lid, secure lid onto base using two small strips of cello-tape
What is step 10 in inoculation?
Place inoculate plate, lid down, into the incubator
When growing in a culture what must the cells get a chance to have?
A gas exchange
What does a typical culture include?
Water, salt, amino acids, vitamins, glucose
What is essential in an animal cell culture medium?
A serum with growth promoting proteins
What is plating out?
It’s when a liquid microbial culture is placed on a solid culture to allow the number of colony forming units to be counted and cell density to be estimated
What is needed to achieve colony count?
Serial dilution
What is a total cell count?
A count of all cells dead and alive
What is a viable cell count?
The number of alive cells that are actively growing/dividing
What is used to estimate the total viable cell count?
A haemocytometer
What is used so cells can be seen when using a haemocytometer?
A vital stain
What is a vital stain?
One that only stains either living or dead cells
Why don’t dies work on most living cell?
The pump them out
What is a common die used in yeast and mammalian cell cultures?
Trypan blue dye (stains dead cells only)
What is a haemocytometer?
A specialised slide that has a counting chamber and a known liquid volume
