How compounds are modified, like if two compounds are modified and used for better adsorption, what will be the chemistry either physical or chemical?

I am doing immunohistochemistry on auditory nerve fibers in the mouse brain. I am perfusing with 0.1% glutaraldehyde, 4% paraformaldehyde to fix the tissue, with the plan of taking confocal images of the fluorescence followed by EM on the same sections. My issue is very high levels of non-specific fluorescence after treating with the secondary fluorescent antibody, and I can't see the structures we expect in the tissue under the confocal. I have heard that this could result from non-specific binding of the antibodies because of free aldehydes, and I have tried treating the tissue with NaBH4 prior to antibody labeling, but cannot seem to decrease the non-specific fluorescence.

Has anyone dealt with this issue or have a protocol for how to reduce it?

What is the accepted punctum size in confocal microscopy for quantification?

Hello,

I have 10nm diameter gold nanoparticle colloids dispersed in the water having 5.7x10¹² particles/ml. can you calculate the molarity of that solution and show the steps? the density of gold is 19.6 g/cm^3.

why fluorescence is highly selective ?

Hello, for a research project, I need to composite graphene oxide with a ceramic powder and sinter it at 1300 degrees under vacuum and argon gas. Does graphene oxide disappear?

If there is a connection between atomic embedding and nano technology,then how far it is useful for quantum mechanics.

What is the correlation between hydrophobicity and zeta potential of metal nanoparticle(s)?

For example, if a nanoparticle(s) has a zeta potential of -30 mV and another has -15 mV, which one would be more hydrophobic than the other?

Appreciate any insight or reference to support the claim!

Thank you.

Hello there!

I am trying to prepare Symbiodinium cultures for microscopy (fluorescence microscopy and potentially confocal). I don't know if i have to fixing my cells in glycolaldehyde (GA) or if i have to fix at all, because I'm trying to capture the chlorophyll fluorescence from my sample, and if I fix it maybe it will die and not be fluorescence anymore.

Could someone help me?

Thank you very much

Hi All, I am using cell mask orange dye to label HEK 293 T cells plasma membrane. I am using 5 ug of dye as final concentration and incubating cells for 10 min at 37•C. I am facing issue with dye internalisation. I did staining before fixation as well as after fixation. In both case All dye internalize. So here is the protocol I am using:

1. Growing cells

2. wash with PBS

3. Stained with cell mask orange for 10 min

4. washing 3 time with PBS

5. Fixation with 4% PFA

6. Washing with PBS.

7. Stored in PBS,Same day Imaging.

Note: Some time I pellet down the cells after second step and then stained the cell suspension. At the end step I just mount the sample on slide.

How can I prevent dye internalization in cell? what are the possible reason of this issue? Please let me know.