I want to estimate uptake of Allexa Fluor 647 labelled nanoparticles by fungi Aspergillus sp. in later stages (26 h, containing hyphae). After incubation in well plates together with NPs, fungi were washed 2 times, fixed and observed (in well plates) by fluorescent microscope. However, background noise is very high and I assume that a lot of particles are still present in the media. Can I centrifuge fungi and observe it on the microscope slides? Or there is another method I can use?
Greetings,
have no experience in this, however, reducing concentration of nanoparticles might help.
Is it possible that labeling agent is in excess? You could check this by observing labeled nanoparticles only before uptake experiment.
Unfortunately, I cannot help. I have no experience in connection with the particular subject matter.
Is centrifugal damage not a consideration? Such damage is well documented in bacteria subjected to excessive centrifugal force. I would be concerned about the accuracy of the uptake after resulting collisions at several times the normal gravitational force.
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