Chris - Week 2: Pilidia and Confocal Microscopy

During week 2, I continued my exploration of pilidial cilia using confocal microscopy. To examine the development of these cilia, I used florescent labeling in an attempt to determine their location on the pilidial surface. This process requires the use of two antibodies: The primary antibody attaches to the protein centrin, which is a component of the basal bodies of cilia (basal bodies can be very roughly thought of as the "root" of a cilium). The secondary antibody, which carries a florescent tag, binds to this primary, and thus reveals the location of cilia to the confocal microscope.                                                          



A confocal micrograph of a pilidium larva of M. alaskensis. The green dots mark the location of centrin-containing basal bodies, and thus the location of the cilia. The white circles are cell nuclei stained with Hoechst, which serves as a positive control.  

As can be seen from the above micrograph, the cilia are typically found to be regularly spaced, and particularly concentrated in the gut and along the ciliary bands. Next week, I will stain different age groups to isolate at which age they begin to develop cilia, and where.