Created
Jul 8, 2024 3:41 PM
Status
In progress
Category
Planaria
- Changing water
- Materials
- Protocol
- Feeding planaria
- Materials
- Protocol
- Preparing planaria food
- Materials
- Protocol
Changing water
Every 2 days and after each feeding
Materials
- Old Tupperware dish
- New Tupperware dish
- Fresh APW (recipe below)
- 100 mL beaker
- Small beaker
- Spray bottle
- Transfer pipette
- Gloves
Artificial Pond Water Recipe (1 L bottle, NIH recipe)
- 500 µL FeCl3
- 2,500 µL CaCl2
- 2,500 µL MgSO4
- 1,250 µL Phosphate Buffer
- Fill to 1 L with DI water
- Label with initials, pH, date, and specify use for planaria only
Because each 6-8 mm planarian requires 1 mL of water, we can place about (source):
- 5 worms in Small Petri dish (35 x 10 mm, 5 mL)
- 10 worms in Medium Petri dish (60 x 15 mm, 11 mL)
- 50 worms in Big Petri dish (100 x 15 mm, 50 mL)
- 100 worms in Extra Small Bowl Ziploc® (9 cm diameter x 5 cm depth, 150 mL)
- 500 worms in Small Square Ziploc® (12 cm x 12 cm x 6 cm, 600 mL)
- 1000 worms in Large Rectangular Ziploc® (15 cm x 26 cm x 8 cm, 1300 mL)
Protocol
- Pour ~250 mL of fresh APW into the new container.
- Pour as much as possible of the old water into the 100 mL.
- If there aren’t any planaria that fell in, dump water down the drain.
- If there are planaria, use a transfer pipette to move them to the fresh container.
- Try not to transfer any food particles/waste when moving the planaria.
- Fill a spray bottle (labelled planaria waste water) with some of the old APW water, and tipping the old container, spray the planaria that stick down to the bottom.
- Repeat steps 2 and 3 until there are no planaria left in the old container and all of them are transferred in the new container.
- If changing after feeding, remove any large chunks of egg yolk from new container
- Label and seal the container with a holed lid cover
This protocol shows cleaning after a feeding (egg leftovers shown) - When cleaning after not feeding, there shouldn’t be any large particles in the old water.
We have multiple populations of planaria which have distinct genetic histories. It is very important not to mix these populations. Change gloves and materials in between populations, if necessary.
Feeding planaria
Once a week
Materials
- Frozen egg yolk
- Spatula
Protocol
- Remove one frozen cube of egg yolk smoothie (around pea sized amount)
- If the cube is broken, scoop enough to fill the tip of the spatula
- Place into the middle of the container
- If needed, break up chunks with spatula
- Allow planaria to eat ~1-2 hrs
- After feeding, change the water (Planaria husbandry - Changing water)
Preparing planaria food
Complete as needed
Materials
- 1000 mL beakers (2)
- Hot plate
- Egg (1)
- Tap water
- Ice
- Aluminum foil
- Paper towel
- Cutting utensil
- Dish
- Spatula
- Red mold
- Freezer
Protocol
Use hot plate in molecular bio suite, 1000 mL beakers found in cabinet above hot plate location. Using more than 1 egg gives plenty of leftovers - freeze any unused product
- Fill beaker 3/4 way full of water
- Place 1 egg into water
- Turn heat on to 10, bring water to boil
Beaker will be hot!
- As soon as water begins to boil, turn off the heat and cover the top of beaker with aluminum foil
- Leave the egg sitting in the hot water for 11 minutes
- As you wait, prepare an ice bath
- Use another 1000 mL beaker, fill 1/2 full with water, fill rest full with ice (found in 4th floor ice chests)
- After the egg sits for 11 mins, drain out hot water, transfer to ice bath, leave the egg submerged for 15 mins
- After ice bath, empty water and place the egg on paper towel
- To peel:
- Tap egg on counter to break shell into small pieces, carefully peel fractured pieces, keeping egg whites intact
- Cut egg in half and scoop out the yellow yolk inside - place to the side in a dish, discard the egg white
- Using cooked egg yolk, smash in a dish with a spatula and a small amount of water
- Mix until it forms a workable paste
- Scoop paste into the red mold and place into freezer for ~2 hrs or until frozen
