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Author: Subject: Recommended Microscope Stains?
entropy51
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[*] posted on 22-10-2009 at 14:18


Quote:
What's a good bacteria to (try to) cultivate for a beginner? Also I'm guessing for an incubator I can use an incandescent light (I looked at egg incubators but strangely they're $$$0


Home Science Tools sells some
bacteria cultures including B. subtilis, B. cereus, E. coli and R. rubrum that are suitable for beginners. Use those to practice your techniques, such as streaking an agar plate to obtain isolated colonies and aseptically transfer. You will also need a wire loop such as Home Science also sells for transferring organisms and streaking plates.

After you have the techniques for isolation down, you can find interesting bugs all around you. Water from a pond that has decaying vegetation in it will usually provide interesting results. So will animal tissues, such as a package of chicken from the grocery store, but you may lose your appetite for chicken :o. They are not sterile! Throat swabs from healthy animals or humans also provide a nice experiment.

You may be able to find a low wattage bulb that will maintain a closed box at about 35 to 37 degrees C for an incubator, but it will be more reliable if you use a thermostat connected in series to control the lamp bulb. The simple thermostat from an aquarium heater can be used.

The CD collection of old Amateur Scientist columns from Scientific American has a couple of incubator designs IIRC.

Read up on aseptic technique and always wash your hands with soap after handling cultures. Wearing latex gloves is recommended. Cultures must be decontaminated before disposal; chlorine bleach works well for this.

[Edited on 22-10-2009 by entropy51]
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chemoleo
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[*] posted on 22-10-2009 at 14:27


On the matter of papain - looks like a neutral pH or slightly acidic is beneficial - see this http://www.jbc.org/content/167/1/199.full.pdf
(if you intend to post more on this, create a new thread and I'll merge it).

Re e. coli - the best source comes from your body - do I need to point out where it grows quite proliferously?
http://en.wikipedia.org/wiki/Escherichia_coli
No need to be touchy about it - see it as it is - science!
Also see the EM images, you can extrapolate the sizes to what you'd see in a visible light microscope.
Definitely, E. coli is best for simple growth - and they can be treated quite badly (centrifuged (compressed), frozen (in 50% glycerol), and revived without a problem)!
Plus their doubling time of 20 min is great - so you won't have to wait for days...




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[*] posted on 6-11-2009 at 17:02


Thanks entropy51 and chemoleo! I finally found some pure Agar yesterday: Had to visit a few Asian supermarkets to find it. The brand I found was Thai (or is that Vietnamese). Anyway $1.90 for 25g which will make 3 litres of the stuff. I'll boil up some beef broth today, mix it with agar and start an E. Coli colony. Thanks chemoleo for the tip. I was was going to ask if I get them from where I think I get them from! :-) I'll also do the chicken one (I already guessed it was swimming in the stuff).

Also my heart was broken a few days ago: Someone was selling an old Olympus microscope with phase contrast on eBay. I bid for it, but someone beat me by a few seconds (and got it for the same price... ARGH! When I woke up the next day the first thing I thought was how I didn't get it. ARGH!!!) Was curious to compare the quality of that with my Indian microscope and *phase contrast* looks nice in the textbooks.

Anyway: Is phase contrast a viable alternative to mucking around with fixers and stains? On my Indian Microscope dark field is nice to 200X but at higher mags it looks just like brightfield. There is an American company selling "microscope phase contrast kits" though it would suck if I bought it and found out it didn't work with my microscope. I'm ordering the stains this weekend. Before I do are there any favorite/useful dyes you would recommend in addition to / amongst these?

Eosin Y, certified C.I. #45380
Methylene blue, certified C.I. #52015
Indian Ink / Nigrosin, w/s C.I. #50420
Basic fuchsin, certified C.I. #42510
Gentian violet C.I. # 42555
Gram's iodine
Safranin O, certified C.I. #50240
Neutral red, certified C.I. #50040
(Will also some PBS)

[Edited on 2009-11-7 by qton]
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[*] posted on 7-12-2009 at 05:49


I am curious about it, maybe I should debug also my yogurt-making process. I think after I do this, I think I will be much satisfied. Making myself or bringing myself too many challenges are my interest.



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[*] posted on 17-1-2011 at 15:25


We should make biology subforum, it could have many interesting threads like those. Keep up with the good work on growing bacteria cultures, I am interested in that too but I am not sure where I will start (I have no available agar). I heard plant tissues can be planted on agar too and using that, we can clone plant from just parts of the leaves.
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