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CROP PHYSIOLOGY & METABOLISM

Multipurpose Cryogenic Surface Apparatus

A Liquid Nitrogen-Chilled Sample Tray

U.S. Water Conservation Laboratory, USDA-ARS, 4331 East Broadway, Phoenix, AZ 85040

* Corresponding author (nadam{at}uswcl.ars.ag.gov)

The use of cryogenic techniques allows collection and storage of large numbers of samples for later processing for biochemical assays. In large field studies it is very difficult to obtain an accurate measure of leaf area as the samples are collected while preserving biological activity. Measurement of leaf area at the time of processing is also difficult because thawing of leaf tissue can cause degradation of leaf biochemical components and processes. The objective of this study was to design a liquid N₂–chilled tray that allows cryogenically preserved samples to be sorted and measured without the risk of sample thawing. A stainless steel lid (working surface) was brazed to a stainless steel pan, creating a reservoir that could be filled with liquid N₂. The working surface could then be maintained at approximately the temperature of liquid N₂. Enzyme activities and polyacrylamide gel separations of ribulose-1,5-bisphosphate carboxylase (Rubisco) and phosphoenol pyruvate carboxylas (PEPCase) of sorghum [Sorghum bicolor (L.) Moench] were used to assess the performance of the tray. Enzyme activity assays and gel separations of Rubisco and PEPCase confirm that biological activity is maintained through use of the multipurpose cryogenic surface (MCS) tray.

Abbreviations: DTT, dithiothreitol • MCS, multipurpose cryogenic surface • PEPCase, phosphoenol pyruvate carboxylase • Rubisco, ribulose-1,5-bisphosphate carboxylase • SDS-PAGE, sodium dodecyl sulfate polyacrylaminde gel eclectrophoresis