Biochemical Society Studentship 2017

Final Weeks I decided to upload just one post covering the final 3 weeks, as they were very similar in that I spent them repeating the macrophage phagocytosis assay and analysing the images I obtained from microscopy. Laboratory Work Macrophage Phagocytosis - Calcofluor White and Concavalin A + FITC Staining This protocol was very similar as I described for week 5, except it involved additional cell permeabilization and second staining steps. Following infection with Candida yeast, samples were stained with calcofluor white, meaning that all external Candida were coloured blue. Then, I fixed the samples and permeabilized the cells before applying the second stain, which was concavalin A conjugated to FITC, which coloured both internal and external Candida green. As concavalin A binds to residues present in the macrophage as well as the Candida , these also became visible with the conA + FITC staining. External Candida Stained with Calcofluor White Ex...

First Week in the Johnston Laboratory My time in the Johnston laboratory will focus on one particular experiment, the macrophage phagocytosis assay. This week I first shadowed Robbie, the post-doc in Dr. Johnston's lab, and then had a go at doing the assay myself. Laboratory Work Macrophage Phagocytosis Assay This experiment aims to determine if and how wild-type and apm4 Δ mutant strains of Candida differ in their interaction with macrophages, a major cellular component of the innate immune system. Macrophages most prominently phagocytose, or "eat", and destroy pathogens in an attempt to protect the body from invading organisms. Due to the fact that: (i) we suspect that the apm4 Δ mutant may have a different cell wall composition in comparison to wild-type, some of the components of which I have been examining over the past 4 weeks (e.g. chitin); and (ii) cell wall components are integral for macrophage recognition of Candida ; this experiment gives a pathophy...

My Last Week Solely in the Ayscough Laboratory This week was my last week in the Ayscough laboratory, as I will be doing experiments in the Johnston laboratory starting from Monday. However, I will still be growing up my Candida with Harriet and would like to squeeze in another cell wall microscopy session if possible; so it is not a permanent farewell just yet. Laboratory Work Experiment 1: Caspofungin Growth Curve Take Two This weeks experiment was almost identical to that of last week, except I used a different concentration of caspofungin, namely 0.2  µg/mL instead of 5 µg/mL. Remarkably, despite the staggered timings and attempting the experiment solo, everything ran smoothly. Unfortunately, my results were not the same as last weeks. Although I got very similar looking trends, these were the reverse of last weeks findings, in that the wild-type strain grew "better" than the apm4 Δ  mutant strain this time around. However, once again none of the differences ...