Juggling mode!

Hi there! Yes I know it, shame on me for such a long silence. But don’t you worry; here I am to catch up with you guys.

Since the last blog post, life in the lab has been pretty hectic. So much work to do and so much still to be done.

Can you imagine how fast does time flow when you are in a lab?

If you don’t pay attention to your timing and follow your daily agenda strictly, you might end up running for the last tube to go home!!!…or waiting for the night bus instead (for hours)!

So, what have I been doing so far? To answer, I have gone through my laboratory notebook and realised I have run quite a few independent experiments between dynamic insulin secretion, resistance of islets to external stress stimuli, receptor activation and generation of intracellular signals coupled with metabolic studies. Ah, don’t forget that we need islets to perform our ex-vivo experiments, thus everything begins with islets isolation from murine pancreases, of course!

A typical week of work can be resumed as follows: [please take something for granted at the moment, I will come back and explain successively]

DAY 1- islets isolation, which means dissecting the organs from a mouse, use some biochemical-based techniques to free the islets from the pancreatic tissues, collect the islets and make them as pure as possible (meaning that we try to reduce the presence of other tissues to the minimum)…then we leave the islets resting over night. Of course islets are not left stranded on a bench but in a special “bedroom” at 37 degrees Celsius, 100% humidity and with a mixture of air that contains 5% Carbon Dioxyde (this is the best condition resembling the body environment – warm, humid and thus happy!). Ah, islets are placed in a nutrient rich culture medium which provides them with all the ingredients to be happy and confy so that they can live and recover from the stress resulting from the isolation

DAY 2- I usually use most of the islets (rougly 60%) for my secretion studies. As you might recall from previous posts, I am a big fan of dynamic secretion, which is technically challenging and time consuming (but addictive!). It takes me about 1-2 hour to prepare all the solutions (the liquids) required for the experiment and another hour to set up the instrument. The secretion itself consists of something like a 2.5 hours run on average, plus another good half a hour to clean the apparatus. I usually generate 1400 samples per assessment and each sample needs to be tested for insulin (in duplicate). It takes roughly 5 hours to set up the insulin assay (between the writing of individual reaction tubes and the set up of the reactions themselves). The measurement of insulin needs 2 days of incubation (meaning that we set it up and let it rest for 48 hours in a fridge) then 4-5 more hours of work to transform it into a (cheerful/sad) number.

 DAY 3- I think you have noticed that DAY 2 is already invading DAY 3 and 4, but yes, I do have a DAY 3 anyway, and I usually set up other experiments such us the response of our islets to stress-related stimuli. In a typical set of experiments I aim to determine if our ligands and/or receptors (signals and antennae, remember) take part to the mechanisms regulating the life span of the islets, protecting them from damage or accelerating the effects of an induced damage. To evaluate this, a defined number of islets is exposed to a cocktail of molecules that are known to promote cell death and we measure the capability of our ligands and receptors to eventually reduce or foster these effects. It usually takes 48h to obtain our valuable numbers, but it is not something that requires continuous monitoring, so just 2-3 hours to begin the experiment, another 2 to terminate it and little more time in-between for the necessary passages proper of the method. So, at DAY 3 we are already juggling around!

 DAY 4- Juggling mode full ahead! There’s the insulin measurement from DAY 2 to be finished, babysitting the experiments from DAY 3 and prepare those of DAY 5, and of course, provide the islets we have in culture with the most confortable environment possible in the mean while (=refresh the culture medium as often as possible – and eventually talk to them to make them feel happier and less lonely!). So, if on DAY 5 we are running any microfluorimetry measurement, for example, we need to set up everything the day before

DAY 5- Assuming we are running a microfluorimetry experiment, between the set up and the run itself, it takes something like 6-7 hours to generate a good number of data to be useful for our scientific needs. Ah, and we do this in a room with dim lights. Do we ever fall asleep??? Shhhhh…don’t tell my boss!

Oh, did you forget we had an on-going viability experiment?! No worries, I did not! It’s done now!

Yes, it sounds complicated and it is, indeed! Because we work with animals, we try to obtain as much data as possible from the lowest amount of biological preparations. We  try not to waste any single islet to keep to the minimum the number of animals we use. Unfortunately, we can’t use any other model at the moment, but we (as well as other labs) are also working on generating valid alternatives to animals for the study of Diabetes (if you interested search for the 3R programs on the internet).

I did not include other things to this agenda, such as data analysis, experimental planning, students tutoring, department committees and reports. Oh, how could I forget the weekly lab meetings (I will dedicate a post to this I promise)!

Do you still wonder why the pubs around the campus are full of crazy people drinking as tomorrow never comes on a Friday evening?!?!
Ya, let’s try not to forget that we are British here (or we pretend to be, at least!).
Friday 5 pm it is drink o’clock!

;-)

{I wonder if Joggling would be the perfect title: a sport that combines juggling and jogging – I don’t think I do really hit my chair so much, indeed!} ;-P

One thought on “Juggling mode!

  1. I don’t even know how I ended up here, but I thought this post was great.
    I do not know who you are but certainly you are going to a famous blogger if you aren’t
    already ;) Cheers!

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