Microbiology Marathon

Do you ever feel that it is just SO ANNOYING if on those rare nights when you go to bed early because you need to get rid of the dark circles under your eyes and a whole well-functioning brain the following day but then you find out yourself can’t fall into a deep sleep or any sorts of those nonsense dreams for about 6 hours?Aha,it almost always happens to me.

So last Saturday ( June the 13th) I had a microbiology practicum.As we all know,microbiology practicum means a day in lab full of mixing and spreading bacteria.So it is obvious that I will need my whole brain to work well and being a sleepyhead is absolutely NOT A GOOD IDEA.Nah due to this importance I forced myself go to sleep earlier the night before.Just for your information,I was lack of sleep last week.I only had 4 hours sleep from Monday to Thursday so I was basically look like a panda with big black circles and my small eyes became even more slanted than they naturally are.I turned off my lights and covered my body with thick blankets,curled in a fetal position with closed eyes but awakened mind for maybe about 6 hours.I don’t know why but my bed is a magical place where I suddenly remember about everything I should have done that day.My nerves system always resists to have a rest when it is needed and well so I spent those 6 hours struggling with it until finally I fell into a nonsense dream at 2 AM.When I got up in the morning,I still looked like a skinny panda and my half lobe of brain was still foggy and full of dreamy atmospheres.

    And just as usual,a series of an unfortunate events happened because of that…
    1)I forgot to bring rubber band to tie my hair (it’s a rule at the lab for girls to tie our hair)
    2)I forgot to bring my lab uniform
    3)I spilled a tube full of e-coli broth and stained my own cloth and legs,I was sterilized by 70% ethanol just like the apparatus ( luckily they didn’t feel it important to put me into the autoclave too LOL JK)
    4)My cheek was stained by malachite green without me realizing it until I got home and screamed at my own reflection at the mirror.(Go Melissa later confessed she actually saw it already but decided not to tell me huh)

Long story short we had microbiology marathon from 7 AM until 5 PM.I did bacteria colorizing,media making,spread plate,pour plate ( this was when I spilled the e-coli broth),streak plate,endospore colorizing,SIM test,SCA test,and MR-VP test.It was a really tiring and busy day but I really enjoyed it though ha ha.I got pretty much new knowledge and experiences and I’m interested in it.

Hmmmhh let’s just see my thousand words…

The equipment for media making,streak plate,pour plate,and spread plate.

The equipment for media making,streak plate,pour plate,and spread plate.

Bunsen,micro-pipette tips ,and beaker glass.

Bunsen,micro-pipette tips ,and beaker glass.

These were the apparatuses.I decided to snap these pictures with my phone to help me studying one day and of course to share them with you hehe I’m such a kind girl,aren’t I?

This was the nutrient gel just before putted inside the autoclave.

This was the nutrient gel just before being putted inside the autoclave.

How to make media for cultivating bacteria :
1)Mix NA suspense by adding 3,6 gr of NA powder ( you can buy them at chemical stores) with 180 ml aquades so the concentration will be 20 gr/l (simple physics)
2)Heat the mixture until boiling for about 1 minute.
3)Close the mouth of the beaker glass with cotton and aluminium foil
4)Put into the autoclave for 15 minutes (121 Celcius and 0,15 MPa)
5)Wait until the mixture is somewhat lukewarm
6)Pour it into petri dishes ( 15 ml per petri dish )
7)Let it cool
Actually there are liquid media and solid media to cultivate bacteria in labs but in this case,I was working with the second one.

Streak Plate Featuring Felix's hand

Streak Plate
Featuring Anita’s hand

How to isolate bacteria using Streak Plate Method :
1)Burn the head of the ose using bunsen ( to kill another kind of bacteria that might be live there,this is just a standard sterilizing step) and draw a cross on the petri dish using pen and number each square
2)Heat the lining of the petri you gonna use
3)Scoop 1 ose of cells and streak it with a zig-zag patern inside the first square,and leave a little tail out of it,at the border of square number 1 &2.
4)Repeat number 1 & 2 and streak the tail you have left before from square 2 until the border of square 3 zig-zagly.
5)Repeat it until square 4 is full.Don’t left any tail here.
6)Leave it in room temperature for 24 hours and you’ll get single bacteria colony in square 4 for result. : )

Tips from me : don’t streak with great pressure or your media will be torn

Spread plate

Spread plate

You just need to spread the cells on the surface of the media using a tool that looks like T tubule.

Nah,I spilled e-coli broth while making this!

Nah,I spilled e-coli broth while making this!

This was the easiest way to make isolate bacteria,but since I’m an anomaly so I turned this easiest part of everything into disaster!You actualy only have to pour the e-coli broth to petri dish then add some NA and last mix it.

SCA test,MR-VP test,and SIM test

SCA test,MR-VP test,and SIM test

Nah

The Geeky Girl Glossary :
1) SIM (Sulfide Indole Motility) to know whether a bacteria is motile or not
2) SCA ( Simmons Citrate Agar) to know whether a bacteria uses citrate as its carbon source or not
3)MR-VP (Methyl Red-Voges Proskauer )to know whether a bacteria produces acetyl methyl-carbynol or not.

And now the bacteria and endospore colorizing…

First ,Crystal Violet Colorizing...

First ,Crystal Violet Colorizing…


then add iodine to enable the bacteria adsorb crystal violet

then add iodine to enable the bacteria adsorb crystal violet


Last do the safranin colorizing and splash it with ethanol .

Last do the safranin colorizing and splash it with ethanol .


Bacillus Subtilis!!!!!

Bacillus Subtilis!!!!!

Excuse the low quality picture.It’s rather looks like an eclipse than bacteria haha .My friend took this photo directly from microscope and it was hard to find right angle for that.

It's me who was holding the plate (the eerie greenie thing was the malachite green which stained my cheek)

It’s me who was holding the plate (the eerie greenie thing was the malachite green which stained my cheek)

It’s endospore colorizing and really this is the MOST DIFFICULT PART EVER. We did this three times to get the green endospore and red cell.

Nahh

Nahh

He was  explaining all those complicated things.

He was explaining all those complicated things.

Well,that’s how I spent my Saturday last week.Hope another adventure is around the corner.

xxo : Joan Exlibris

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