Week 5 - Ben Grodner

I can’t believe how quickly time is moving. Summer is getting into the dog days of heat and humidity, and even a short walk to the lab gets sweat glands pumping. My lab work this week involved a fair bit of tissue slide preparations. We have sectioned PJI tissue samples and I am staining for microbial content. Now that I’m settled in the lab space, its possible to move quickly through the various steps without pauses to ask silly questions like “Where are the extra pipette tips?” I owe much to my lab mates for helping me get oriented by dealing with constant questions. 

During my preps I had the chance to sit down with a histopathologist in Dr. Rodeo’s lab and learn some the “artistry” involved with sample fixation and sectioning. It turns out that a good slide prep comes out of experience making bad slide preps and optimizing for the specific sample. As such, I was not terribly surprised when my slides had various issues once I got to imaging them. I spent quite a bit of time troubleshooting for autofluorescence and trying to optimize the imaging conditions. Through it all, I gained some good experience and I’m motivated to optimize further next week. 

Synovial tissue from a combined chronic and acute prosthetic joint infection stained with DAPI (blue) for double stranded DNA (nuclei) and EUB338 single-stranded oligonucleotide hybridization probe (green) targeting 16s rRNA in bacterial species.

This week I also shadowed Dr. Andy Miller, a reputed infectious disease clinician from HSS. He took me on his rounds to inpatients who had developed infections after surgery. When infection does occur after surgery, which is not often, it is terrible for the patient. Patients have to go on intensive antibiotic courses for a long time, and often there are other complications. Dr. Miller showed me how the microbiology lab data was used to inform treatment plans, and then how he would talk to patients in this difficult situation. Interestingly, he showed me how well-read patients are often harder to treat because they second guess themselves and are less straightforward to deal with than those who just report how they are feeling. We had a great discussion on the positives and negative of current diagnostic techniques as well as the gaps in our understanding of chronic and acute infection. Culture techniques are fundamentally limited in the variety of microbes they can identify, but they are incredibly powerful for the functional information they supply. In post-surgery infection, Staphylococcus species account for the vast majority of cases. These species are largely identifiable by culture techniques and important information can be gleaned from culture on their origin (i.e. skin, or other) as well as susceptibility to antibiotic treatment. Although culture techniques are powerful, they are essentially the only tool available for diagnosis and in the hospital. Great work is being done, for example in the Donlin lab, trying to advance next-gen sequencing diagnostics and other molecular techniques, but the issue with this as well as my project is the lack of functional information and trust by doctors. 

I think my research has the potential as a tool for understanding the progression of infection in a more direct way than culture or next generation sequencing. Culture especially is a very removed readout on the actual issue, so I hope that a tool for a more direct observation of the system (still in situ, not in vivo unfortunately) will yield some useful information. However, my time with Dr. Miller showed me how difficult the problem is and how much clinicians rely on well established culture techniques. It is incredibly challenging to break into a field with such a powerful tool, even if it is limited.