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When in the presence of 3.00 um beads, or 6.00 um, or 10,00 um beads, how many bead filled vesicles will Tetrahymena internalize?

  • surplus beads
  • several exploded

not as many beads internalized

Lab 4: Phagocytosis of differently sized beads in Tetrahymena pryfomis

Design

Question

How does the size of a

particle affect phagoctosis

Experimental Question

Procedure

Analysis

Figure 1:Tetrahymena pyrifomis internalizing 3.00 um, 6.00 um and 10.00 um beads. A shows a Tetrahymena with 5 vesicles filled with 3.00 um beads. This tetrahymena was in the presence of about 0.027% of 3.00 um beads (about 1.075 x 10^6 beads).The picture is at 400x magnification and the black bar represents 12.5 um. Each small tick on the ruler in the shot is 2.5 um. B shows a tetrahymena with 4 6.00 um bead filled vesicles. This tetrahymena was in the presence of about 0.027% of 6.00 um beads (about 1.340 x 10^5 beads). This picture is at 100x magnifcation and the black bar represents 50 um. Each small tick on the ruler in the shot is 10 um. C show a tetrahymena that was in the presence of about 0.027% of 10.00 um beads (2.912 x 10^4 beads). It has not internalized any bead filled vesicles. The blue circles outside of the Tetrahymena are the beads. This picture is at 100x magnification and the black bar represents 50 um. Each small tick on the ruler in the shot is 10 um

  • Mix 25 uL live Tetrahymena with 25 uL of 3.00 um beads in a microcenterfuge tube.
  • Observe for 20 minutes
  • Mix 25 uL live Tetrahymena with 25 uL of 3.00 um beads in a microcenterfuge tube. Wait 5 minutes then fix with 25 uL of 3 % gluteraldehyde
  • Observe
  • Add 20 uL 3.00 um beads with 20 uL tetrahymena in a microcenterfuge tube. Wait 10 minutes then fix with 20 uL 3% gluteraldehyde
  • observe
  • Repeat previous 2 steps with 6.00 um beads. observe
  • Repeat previous step with 10.00 um beads.

B

A

C

So...

  • More 3.00 um bead filled vesicles than 6.00 um beads, and more 6.00 um then 10.00 um (no 10.00 um beads were internalized).
  • More 3.00 um beads internalized than 6.00 um beads, and no 10.00 um beads internalized.
  • Problems
  • comparable?
  • range

Figure 2: Average bead-filled vesicles in Tetrahymena pyriformis internalized based on bead size. Average drawn from 40 individual Tetrahymena pyriformis. Error bars represent standard deviation.

Jennifer Guzman, Maya Martin-Udry, Jessie Lu, Kristin Williams

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