A portable cryo-plunger for on-site intact cryogenic microscopy sample preparation in natural environments.
We present a modern, light portable device specifically designed for environmental samples for cryogenic transmission-electron microscopy (cryo-TEM) by on-site cryo-plunging. The power of cryo-TEM comes from preparation of artifact-free samples. However, in many studies, the samples must be collected at remote field locations, and the time involved in transporting samples back to the laboratory for cryogenic preservation can lead to severe degradation artifacts. Thus, going back to the basics, we developed a simple mechanical device that is light and easy to transport on foot yet effective. With the system design presented here we are able to obtain cryo-samples of microbes and microbial communities not possible to culture, in their near-intact environmental conditions as well as in routine laboratory work, and in real time. This methodology thus enables us to bring the power of cryo-TEM to microbial ecology.
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA. lrcomolli@lbl.gov
Luis R
LR
Comolli
Robert
R
Duarte
Dennis
D
Baum
Birgit
B
Luef
Kenneth H
KH
Downing
David M
DM
Larson
Roseann
R
Csencsits
Jillian F
JF
Banfield
D020285:Cryoelectron Microscopy / Q000379:methods*; D004783:Environmental Microbiology; D005615:Freezing; D019095:Point-of-Care Systems; D013048:Specimen Handling / Q000379:methods*