Nuffield Day 17

With the data for the bearings, I now had to export to excel and analyse.

The extra data has confirmed my previous conclusions that there is a the Golgi has a bias towards the leading edge – showing cell polarity.

Nuffield Day 16

With all the images taken for the third embryo on the confocal, it was now back to Volocity (a computer programme). I analysed each image taken of the sections of the palate ranging from anterior to posterior. I drew lines from the Golgi to the Nucleus and the bearing was measured. I managed to complete this for all sixteen images!

Nuffield Day 15

Back after the bank holiday, I was booked in for the confocal for the whole day. Eleni helped me to image palates for a third embryo. It was really interesting and amazing how the confocal worked with lasers to produce the image.

Nucleus stained with DAPI (blue), Golgi stained with Anti-rabbit-488 and as the mouse used was a tomato cross, the membranes are stained red.

Nuffield Day 14

The confocal was booked early for a couple of hours again. This time, with cranked up the laser, to produce clearer images. Four more images were produced, so five in total so far.

After the session, I began to draw lines to measure the bearing between the Golgi and the nucleus straight away for this embryo C.

After the bank holiday weekend, we have booked the whole day on the confocal microscope so hopefully we will get all the images completed, so I will be able to complete my data analysis and project.

Nuffield Day 13

I completed my analysis of the bearing of the Golgi body for embryo A and B including placing them in zones. There has been a clear relationship, showing that developing palatal cells have cell polarity, with the Golgi at the leading edge.

With the prepared slide, I began to image, using the Confocal microscope. The confocal microscope uses powerful lasers to highlight each antibody stain which are merged. With only a couple of hours booked, we managed to take one image successfully, however it was not as clear as we would have liked.

Nuffield Day 12

I continued to update my project with difficulties  had encountered. I also met with Jeremy, my supervisor, before he left to go on holiday.

Eleni helped me to prepare the third specimen, preparing the sections with antibody stains and keeping cool overnight.

Nuffield Day 11

I made radar plots of the data and made sure that a bearing between the golgi and nucleus of 180 = the direction of growth. I also drew up an outline for the project.

In the afternoon, I washed slide with sectioned palate, added blocking solution, then the antibody stain for the Golgi body to be left overnight in the cold room.

Nuffield Day 10

Monday’s generally begin with a lab meeting, where we dicussed how everyone’s project was moving along. I met with my supervisor to clarify some data and on the my overall project. I continued to collate the data in different ways, putting them in zones. Now all the data is relative to it’s principal axis, so each image can be compared fairly. Noticeable patterns between golgi orientation and direction of growth are clearly showing.

A few labs combine together for a presentation every few weeks, Eleni from our lab presented about a recent project of hers on the development of teeth.

Nuffield Day 9

After sorting out data on excel, I got to do some lab work using the Cryostat. The cryostat cuts frozen sections. I cut 30micron section of E13.5 mouse palates, I managed to get good sections but the centre of the palate did not come up well enough. We have two more embryos, a friend kindly sectioned for Monday morning.

Nuffield Day 7

I started by following Carly to help her extract mouse embryos at E13.5, it was fascinating how each embryo has their own sac, but all are joined together in a line looking much like beads. I further worked on my project writing about some difficulties I have already encountered and some methodology. Eleni showed me her project with xenopus, looking down the microscope at their divisions. She also helped me to change the solution that the embryo I will be using was in. This is part of the preparation in order to image the palate.