We culture cells in 8 chamber slides and do immunofluorescent staining. We put one drop of mounting media on each of the 8 wells and drop the coverslip at a 45 degree angle. Does anyone have tips on how to get rid of air bubbles trapped between the coverslip and the slide?
Dear Helen,
My suggestion is: Do not put one drop on each of the 8 wells, but put a continuous strip along the center between the 4 wells to the left and to the right, and then let the cover slip very slowly lower down from one end to the other end. This allows the mounting media dodge to both sides without forming bubbles. Then remove the excess mounting media on the sides of the coverslip. Just in case: if the cover slip is damp from the humidity in the room, please remove the humidity with a small gas flame before mounting.
Good luck!
Best regards,
Guenter
Dear Helen,
My suggestion is: Do not put one drop on each of the 8 wells, but put a continuous strip along the center between the 4 wells to the left and to the right, and then let the cover slip very slowly lower down from one end to the other end. This allows the mounting media dodge to both sides without forming bubbles. Then remove the excess mounting media on the sides of the coverslip. Just in case: if the cover slip is damp from the humidity in the room, please remove the humidity with a small gas flame before mounting.
Good luck!
Best regards,
Guenter
In addition to carefully move when lowering the cover slip. we dilute the mounting medium witch makes it harder for bubbles to form. We dilute adding 30% more of xilol to enthelan. if you press down the cover slip after mounted you can expel the formed bubbles if they are present.
Best wishes
Dear Helen,
If you are using an oil or glycerol-based (non-drying form) mounting medium, and if the bubble reaching the edge of the coverslip then just add more mounting medium to that edge and the medium will be sucked under. If the bubble is right in the middle and not reaching the edge than try to get it to reach the edge with minimal pressure only on one side move it closer to the edge of the coverslip edge, once reaches the edge again add more mounting medium and as it will be sucked under the bubble will move out replaced by the medium. When applying the medium use a pipette (10-100 microliter) with a yellow tip so that when you add the media gentle and in one point, to allow space for the bubble to escape.
best wishes, Refik
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Histological Techniques