Isolating Single Cells from Phytoplankton

The Flow Cytometer I was going to use

The Flow Cytometer

One of the normal things people say about science, is that things almost never goes by the way you expect them to, that behind every project there are many backup plans, and I’ve proven this to be right. For the last few weeks I have been growing cultures of six different phytoplankton, and running fluorescence test on each one of them to keep track of their growth phase: exponential or stationary. With the intention of isolating single cells from each culture and do the qPCR and cell cycle analysis. This way I would be able to know the rRNA gene content number. Plan A was to isolate single cells using the flow cytometer: a machine that analyzes single cells, making them pass through a laser and using fluorescence signals to sort the cells and separate them into different properties using light.

So many Alexandium cells moving around!

So many Alexandrium cells moving around!

Still, the flow cytometer wouldn’t work when I tried to use it. Therefore I had to go with plan B: isolate singles cells from one of my phytoplankton cultures with a pipette. Meaning, I spent a day, in front of a microscope, isolating 78 single cells from Alexandrium (the phytoplankton I ended up choosing). After isolating the cells, I ran qPCR analysis on them. This was made today, and it appears to be successful, still the data needs yet to be analyzed to see how it turned out. Hopefully it will turn out the way I expect it to, and even better, we manage to fix the flow cytometer.

 

There is ONE cell in each of this PCR's wells!

There is ONE cell in each of this PCR’s wells!

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