Microscopy

[Caliban07]

Apologies if this is in the wrong thread. I've also sought advice from someone on here about this topic and would like to let them know their advice was fully appreciated and adequate. Unfortunately I have lost the messages and would now like to pose the same question to more people.

What types of microscopes will allow me to identify bacteria so I can diagnose disease. What magnification would be required and what techniques would have to be used to observe.

Same question for parasites.

Thanks

 

[Delapool]

Are you planning to stain for gram negative or positive bacteria?

 

[Caliban07]

Are you planning to stain for gram negative or positive bacteria?

Yes that's the aim.

 

[Delapool]

Texted my sister but long shot. Will let you know if I hear anything.

 

[Caliban07]

Thanks. Spoke to a pal of mine who's wife is a microbiologist and she has recommended a scope for me. Said she will even run a few tests for me at work if I need to confirm anything

 

[Mebbid]

I believe it was me you were talking about with this. You can only really identify a few things about bacteria when using anything but electron microscopes. Even then, you still won't readily be able to differentiate between disease causing bacteria and normal bacteria in the tank.

For parasites, generally you have to perform a dissection of the affected area to get a sample to look at. However I am not sure if a standard light microscope would work for this.

 

[Caliban07]

I believe it was me you were talking about with this. You can only really identify a few things about bacteria when using anything but electron microscopes. Even then, you still won't readily be able to differentiate between disease causing bacteria and normal bacteria in the tank.



For parasites, generally you have to perform a dissection of the affected area to get a sample to look at. However I am not sure if a standard light microscope would work for this.

I'm confused. I asked a microbiologist and she didn't say it couldn't be done. I gave columnaris as an example and told her I only wanted to identify the bacteria.

This is what she said:

Lex says : You would need microscope with a x100 lens and you would need to look under oil emersion. Best stain would be a gram stain: Crystal violet then iodine then acetone alcohol and finally saffranin. This is the most basic method you can do. What you up to?

And

You can see parasites under much lower magnification, depending on the parasite size. Usually either x20 or x40 and parasites usually in much lower numbers so you would need to look on low powered fields so you don't miss them. Our best microscope at work is an Olympus, it's fluorescent too, but don't think you'd need that unless u try and stain for parasites. Just had a look at second stage tables for flexi and it says it shouldn't grow on nutrient agar but will grow on blood with neomycin agar, and it grows cold. We use both that agar if u want some? You would need to settle or centrifuge the water, or swab the fish?? Bit out of my depth! Lol

 

[Mebbid]

I'm confused. I asked a microbiologist and she didn't say it couldn't be done. I gave columnaris as an example and told her I only wanted to identify the bacteria.

This is what she said:

Lex says : You would need microscope with a x100 lens and you would need to look under oil emersion. Best stain would be a gram stain: Crystal violet then iodine then acetone alcohol and finally saffranin. This is the most basic method you can do. What you up to?

And

You can see parasites under much lower magnification, depending on the parasite size. Usually either x20 or x40 and parasites usually in much lower numbers so you would need to look on low powered fields so you don't miss them. Our best microscope at work is an Olympus, it's fluorescent too, but don't think you'd need that unless u try and stain for parasites. Just had a look at second stage tables for flexi and it says it shouldn't grow on nutrient agar but will grow on blood with neomycin agar, and it grows cold. We use both that agar if u want some? You would need to settle or centrifuge the water, or swab the fish?? Bit out of my depth! Lol

Yep sounds about right.

What I mean is, you will be able to see some characteristics of bacteria, but identifying them is next to impossible.

For example:
It can be difficult to find specific things but here are some I picked out:

E. Coli - Everywhere and commonly studied:

E. Coli 1000x

E. Coli under scanning electron microscope

Vibrio - Can be found in aquariums
Vibrio under scanning electron

Theres a crap load of other types of bacteria that look just like both of these. There's really no species specific identifying characteristics to look for to differentiate between different types of bacteria.

Parasites are much easier to identify but harder to find hence my statement about dissection.

 

[Delapool]

My sister said similar microscope it sounded but said a vet might be able to do it.

 

[pip walters]

You should see all common ectoparasites wit x20 buddy. That's what I use.

 

[Caliban07]

Thanks but I'm more concerned with the bacteria.