Em 208 microscope

We performed transmission electron microscopy (TEM) as previously described^{18 (link)}. The tissue is fixed in MJK solution and rinsed three times for fifteen minutes each in a 0.1 M Sodium Cacodylate buffer. The tissue is then placed in a 1% osmium tetroxide for 1 h. The tissue is then rinsed with 50% ethanol, and then sequentially dehydrated in 50% ethanol, 75% ethanol, and 95% ethanol for 15 min each. Lastly, the tissue is placed in 100% EtOH (15 min × 2). Next, the tissue is placed in propylene oxide for (15 min × 2). Then the tissue was preinfiltrated with a 2:1 propylene oxide resin for 1 h, then 1:2 propylene oxide resin for 2 h, and then pure resin for 1 h. Lastly, the tissue is embedded and polymerized at 60 degrees C overnight. The embedded tissues were sectioned with a Leica EM UC6 ultramicrotome at a thickness of 90 nm and collected on copper mesh grids. The following day, the tissue is stained with alcoholic uranyl acetate for five minutes and then Reynolds lead citrate for five minutes. TEM imaging was performed on a Philips EM 208 microscope at 60 kV using a Hamamatsu digital camera.

We performed transmission electron microscopy (TEM) as previously described^{18 (link)}. The tissue is fixed in MJK solution and rinsed three times for fifteen minutes each in a 0.1 M Sodium Cacodylate buffer. The tissue is then placed in a 1% osmium tetroxide for 1 hour. The tissue is then rinsed with 50% ethanol, and then sequentially dehydrated in 50% ethanol, 75% ethanol, and 95% ethanol for 15 minutes each. Lastly, the tissue is placed in 100% EtOH (15 minutes × 2). Next, the tissue is placed in propylene oxide for (15 minutes × 2). Then the tissue was preinfiltrated with a 2:1 propylene oxide resin for 1 hour, then 1:2 propylene oxide resin for 2 hours, and then pure resin for 1 hour. Lastly, the tissue is embedded and polymerized at 60 degrees C overnight. The embedded tissues were sectioned with a Leica EM UC6 ultramicrotome at a thickness of 90 nm and collected on copper mesh grids. The following day, the tissue is stained with alcoholic uranyl acetate for five minutes and then Reynolds lead citrate for five minutes. TEM imaging was performed on a Philips EM 208 microscope at 60 kV using a Hamamatsu digital camera.