Microscrope gel slide

From FreeBio

The idea is to make a thin sheet of gel in between a large microscope slide and a small cover glass. (large microscope slide: 25 x 75 mm, small cover glass: 22 x 22 mm) Don't make the slides too long before you are ready to look at your samples because they dry up and crack if they have been left out too long after being made. Making them half an hour before you are ready to visualize would be ideal.

The gel is 2% low-melt Agarose dissolved in M9 medium, you need around 300µL of gel per slide.

1. Mix the M9 media and low-melt agarose (to make a 2% mixture) and microwave until the agarose is dissolved.
2. Let the mix cool until you can handle the container in which you heated it up.
3. Put a small drop of gel solution on top of a big microscope slide (how much gel solution? you do it by eye, I never use more than 300µL. You need to put enough down so that when you drop a small cover glass on top of the gel mix it will expand under it and ooze a little, but not too much, from the sides of the small cover slip.)
4. Drop a small cover slide flat on top of the gel drop in the big slide. This step takes some practice so that the cover slide lands and doesn't float off croockedly off the slide.
5. Let the gel cool. It shouldn't take more than ten minutes.
6. To load your sample you need to:
   • Gently remove the coverslip using one of the gel extraction blades.
   • Use the same blade to remove any of the gel that might of oozed out from the sides of the cover slip and is higher than the gel hat dried under the cover slip, just cut the gel and scrape it away.
   • Load your sample on top of the gel and put the cover slip on top of that. You now have a gel sample sandwhich in between the two slides.

Another option, instead of these slides is to use the "3cm plate preparation method" a.k.a. "The little plate method" pioneered by Mr. Yin Li. In this method you make little LB-agar plates (little as in 3cm in diameter), put your sample on top of it (about 20µL), and on top of that drop a cover slip. The advantages of this method are that the little plates are less likely to dry out, the baceria will be happier in them, and they are easier to make.

140.247.92.120 12:12, 12 Jul 2005 (EDT)noriega