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Confocal - Microscopy Imaging Techniques All about Microscopy, Confocal Microscopy, Fluorescence microscopy and Other Imaging Techniques


Microscopy - LR White resin and TEM

Microscopy - LR White resin and TEM - Confocal - Microscopy Imaging Techniques

Microscopy - LR White resin and TEM - All about Microscopy, Confocal Microscopy, Fluorescence microscopy and Other Imaging Techniques


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Old 08-24-2011, 05:15 PM
Pipette Filler
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Default Microscopy - LR White resin and TEM



Hi,

I'm using LR white resin for the first time and just stained some ultrathin sections on nickel grids with 2% aqueous uranyl acetate and lead citrate and tried to look at them on the TEM.

The sections seem to disintegrate in the beam very quickly, even after I have made sure the beam is low and changed grids with the microscope set to low mag and beam. This applies to all parts of the section, even ones not directly aimed at by the beam, as when I scan around new areas already seem to have shrunk towards the grid bars and become crumpled up against them.

I have only seen this happen before when using SPURR resin when the beam has been too high in areas directly hit by this beam, especially those weakened by hole/knife marks.

Any suggestions?

Thank you!
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Old 10-06-2011, 11:01 AM
Pipette Filler
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Default Re: Microscopy - LR White resin and TEM

Hi PetraK

There are lot of reasons for this...

First of all, how do your ultrathinsections look like when you are cutting them? Do they have holes or do they disintegrate on the water while cutting?
How thin are your sections regular 80 nm or are you doing thinner sections for EELS etc?

I would assume that you have a problem with polymerization:
* so I would make sure that you really follow the guidelines for LR white polymerization. * Make sure that there are no airbubbles within LR white before polyermization
* Make sure that the specimen is on the bottom side of the mold you are using for polymerization
* Or could it be the case that your LR white is expired?
* when everything fails, switch to LR Gold...


I only made such experiences with LR white or LR Gold when something went wrong with the polymerization.

Further it is possible that sections are "polluted" with heavy elements which absorb a lot of electrons, which heats up the section, what further leads to disintegration. Maybe check your waterbatch on the knife (and never use these filthy reservoir pumps of the microtoms...)

best wishes from Innsbruck
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microscopy , resin , tem , white


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