You raised some very good points! Let us comment on some of them:
To ensure low background signal using silicon gaskets, two points are important:
1) make sure there is no air between the gasket and the coverslip to prevent leakage. This is easily achieved by applying some pressure from the other side of the glass with tweezers or a clean pipette tip, without touching the surface in the wells. Check this video to see it in action https://www.youtube.com/watch?v=tWplRYgaGRI.
2) After use, discard the coverslip and store the used gaskets in isopropanol (up to a few weeks). To clean the gaskets: discard the storage isopropanol, add MilliQ or similar water and shake vigorously. Discard water and repeat with isopropanol, then water again. This can be repeated if the result is not satisfactory. Finally, dry the gaskets with a kimwipe or similar tissue, and store in a dry environment until use. Important: Discard the gaskets if they come in contact with detergents.
With these tips, we do not see significant background differences between silicon gaskets and flow chambers. Would love to hear your experiences, though!
Hi Jason,
You raised some very good points! Let us comment on some of them:
To ensure low background signal using silicon gaskets, two points are important:
1) make sure there is no air between the gasket and the coverslip to prevent leakage. This is easily achieved by applying some pressure from the other side of the glass with tweezers or a clean pipette tip, without touching the surface in the wells. Check this video to see it in action https://www.youtube.com/watch?v=tWplRYgaGRI.
2) After use, discard the coverslip and store the used gaskets in isopropanol (up to a few weeks). To clean the gaskets: discard the storage isopropanol, add MilliQ or similar water and shake vigorously. Discard water and repeat with isopropanol, then water again. This can be repeated if the result is not satisfactory. Finally, dry the gaskets with a kimwipe or similar tissue, and store in a dry environment until use. Important: Discard the gaskets if they come in contact with detergents.
With these tips, we do not see significant background differences between silicon gaskets and flow chambers. Would love to hear your experiences, though!
Additionally, for faster coverslip cleaning, the batch procedure can save some time (https://www.youtube.com/watch?v=Mq_X4VkNNxo). The rack shown in the video can be found here https://www.emsdiasum.com/microscopy/technical/datasheet/72243.aspx