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Bioluminescence vs Fluorescence Microscope

bioluminescence fluorescence microscope

Best Answer bob1, 14 December 2017 - 06:50 AM

Your confocals should have the ability to take white-light pictures, just use it as if you were imaging a cell under normal illumination.

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#1 Thomson

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Posted 13 December 2017 - 05:40 PM

Hi, I'm planning to observe the protein expressed in living cells using Nano-Glo live cell assay system. It is a bioluminescence substrate.

 

However, my lab only have 2 fluorescent microscopes, which are:

1) Light Sheet confocal microscope (SP8) Leica Microsystems

2) Confocal laser scanning microscopy (FV1200-IX-KU) Olympus

 

So my question is: Since luminescent microscope emit light by itself, can we just use fluorescent microscope to view it (without emit any wavelength to excite the reporter)? Or our lab have to buy a bioluminescence microscope?

 

ps: I've googled online on the above microscopes but they did not state its use regarding bioluminescence imaging.


Edited by Thomson, 13 December 2017 - 05:42 PM.


#2 bob1

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Posted 13 December 2017 - 06:55 PM

Assuming that there is enough signal from the luminescence, it should be fine to use those microscopes. To do this you don't need the confocal/fluorescence capabilities - you would just be using them as a regular microscope. I am guessing (having never done this application) that you would need both low f stop lenses and a decent camera (for long exposures), both of which are usually found in confocal systems anyway.



#3 Thomson

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Posted 13 December 2017 - 11:16 PM

Assuming that there is enough signal from the luminescence, it should be fine to use those microscopes. To do this you don't need the confocal/fluorescence capabilities - you would just be using them as a regular microscope. I am guessing (having never done this application) that you would need both low f stop lenses and a decent camera (for long exposures), both of which are usually found in confocal systems anyway.

 

I see. I'd try it first as you suggested. I am just worrying if the filters in the microscope would block some/all of the luminescence signal. Thanks!



#4 bob1

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Posted 14 December 2017 - 06:50 AM   Best Answer

Your confocals should have the ability to take white-light pictures, just use it as if you were imaging a cell under normal illumination.



#5 Thomson

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Posted 20 December 2017 - 06:06 PM

Your confocals should have the ability to take white-light pictures, just use it as if you were imaging a cell under normal illumination.

 

Hi, I forgot to ask, similarly, for flow cytometer, it should be the same thing right? I just make sure to turn off fluorescence laser beam so the machine should still be able to detect bioluminescence substrate?


Edited by Thomson, 20 December 2017 - 06:21 PM.


#6 bob1

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Posted 21 December 2017 - 08:43 AM

I'm not sure on this one - it may depend on the type of machine you have and the sensitivity of its detectors. There are a number of papers on the topic, so I suppose that it is possible, but may not be feasible with the instrumentation you have. I would talk to the technical support you have at your institute and probably to the FACS company technical support as well.







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