Friday, 25 October 2013

Observing and genotyping C. elegans

I had a fun day yesterday doing the "C. elegans" practical, which is part of the first-year labs. In the practical, we got to look at live C. elegans and also set up some PCR reactions to genotype some wild-type and mutant C. elegans. It was all fun stuff.

Below is a movie we shot of the worms on the day of the practical and some stills from the microscopes.

DA230017

DA230028

DA230027

DA230026

DA230024

If you would like to support my blogging efforts, then please feel free to buy me a coffee at https://www.buymeacoffee.com/drnickm

Sunday, 13 October 2013

Fun and games with Paramecium

Last week we had our annual first-year 'paramecium practical'. The aims of the session were to familiarise the students with the microscopes and to also introduce unicellular organisms.

In the UK we order the paramecium for the practical from a local supplier. However, in Malaysia, despite 4 months of searching, I was unable to find a supplier. This left me with three options:

  1. Don't do the practical
  2. Find an alternative supplier
  3. Culture my own…

I tried to find an alternative supplier (paramecium are used to feed baby fish), but none of the local pet stores stocked them - although I did get a rather nice bag of water fleas (Daphnia).

IMG 5124

A bag of Water Flea - Daphnia

D9290027

A Water Flea - Daphnia

Finally, I decided to culture them.

After a quick read around the Internet, I found a recipe for culture media and some advice on where to find paramecium.

The general advice for 'hunting' paramecium is to take water from close to the top or the edge of a pond. In the end, I took a sample of water from the surface, plus some material from the bottom of the pond. This was allowed to settle overnight (see photo below).

The culture media consisted of dog biscuits, and 4 or 5 cm2 of lettuce leaf, in water.

IMG 5128

Dog Biscuits

IMG 5133

Lettuce Leaf

IMG 5127

Water

IMG 5134

Pond Water - a mixture of material from the bottom of the pond, and water from the surface and edge

IMG 5136

Two culture bottles and a bottle of bond water

I set up three bottles, two culture media, and a bottle of pond water. These were left overnight.

And the next morning… The pond water had cleared, and the bottles with dog biscuits and lettuce were cloudy (the cloudiness was caused by bacteria growing) - these will be a food source for the paramecium.

One of the 'culture media' bottles was seeded with 20 ml of water from the top of the pond bottle, and the other with 20 ml of water collected near the vegetation at the bottom of the pond bottle.

IMG 5138

Next morning - the three bottles…..

An examination of the pond water under a microscope revealed the presence of paramecium, but not that many.

D9290007

There were also some other ciliates present…

D9290038

Over the next 12 days, the cultures got progressively more cloudy and exceedingly smelly.

IMG 5141

12 days after adding the pond water

Of the two culture bottles, the one seeded with water from the top of the pond bottle produced the best (most and largest) paramecium, and the best place to harvest the paramecium was from the top 5 - 10 mm of the bottle, and in particular the layer of 'scum' that had formed on the top of the water. Paramecium taken from elsewhere in the bottle tended to be smaller.

DA110091

Movie

The movie below was shot during the practical.

If you would like to support my blogging efforts, then please feel free to buy me a coffee at https://www.buymeacoffee.com/drnickm