Re: [DIYbio] Camera for fluorescence detection system

My lab was able to visualize fluorescence (protein of interest) in cells working from some of the instructions found here (Haseloff lab, Cambridge).


https://webcache.googleusercontent.com/search?q=cache:GbEcbAud3GsJ:https://haseloff.plantsci.cam.ac.uk/imaging/cheaposcope/optics/index.html+&cd=11&hl=en&ct=clnk&gl=us
On Wednesday, December 9, 2020 at 2:08:41 PM UTC-8 John Griessen wrote:
On 11/25/20 4:31 AM, 'Charles Maldonado' via DIYbio wrote:
> " Is the light from one particle varying a lot if it is against the far wall of the tube or near wall?"
>
> Unfortunately, I don't know, whether the light is varying or not depending on the position of the particle.


Hi Charles, I asked trying to guess what your goal is -- maybe to separately count particles without counting two as one.


> "Will there be a low low chance of two touching or close to each other?"
>
> I think, the chance will be quite high...

That makes the goal of count-particles-without-counting-two-as-one difficult.


> "Are you wanting to do any hardware or code development, or just "full manual" proof of concept?"
>
> Some code <snip> My initial thought was getting the intensities of my cameras pixels, calculating the average value, normalizing it to some kind of maximum and then plotting it.

But which pixels would mean what if you don't have control over the place they are aiming at, or whether the particles are in that
zone? This seems to be that same goal as is called flow cytometry, which some have charge big bucks for and there are probably
some patents involved if any gear is built for hire. Can you think of anything easier as a goal? It does sound like you want a
microcontroller to read particle sensors and save data to send along when you ask it.

Can the flow rates be very slow and work? What liquid volumes can you pump in a volumetric controlled way?
ONe way to spread the particles out is to make a drawn glass tube thin enough in the middle to let just one or so pass, and keep
the concentration of the suspended cells/particles low enouigh that they don't all rub against each other -- plenty of
interstitial liquid -- so the volume of liquid spreads them out as they go through the narrow drawn part of the tube.

Sounding like what you were thinking yet?

--
John Griessen
industromatic.com Austin TX building lab gear for biologists

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