Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 7-Day Trial for You or Your Team.

Learn More →

A Stopped‐flow/rapid‐freezing machine with millisecond time resolution to prepare intermediates in biochemical reactions for electron microscopy

A Stopped‐flow/rapid‐freezing machine with millisecond time resolution to prepare intermediates... We have developed an instrument capable of freezing transient intermediates in rapid biochemical reactions for subsequent freeze‐fracturing, replication, and viewing by transmission electron microscopy. The machine combines a rapid mixing unit similar to one widely used in chemical kinetics (Johnson, 1986) with a propane jet freezing unit previously used to prepare static samples for freeze‐fracturing (Gilkey and Staehelin, 1986). The key element in the system is a unique thin‐walled flow cell of copper that allows for injection and aging of the sample, followed by rapid freezing. During freeze‐fracturing, a tangential cut is made along the wall of the flow cell to expose the sample for etching and replication. The dead time required for mixing and injection of the reactants into the flow cell is less than 5 ms. Electronic controls allow one to specify, on a millisecond time scale, any time above 5 ms between initiation of the reaction and quenching by rapid freezing. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Microscopy Research and Technique Wiley

A Stopped‐flow/rapid‐freezing machine with millisecond time resolution to prepare intermediates in biochemical reactions for electron microscopy

Download PDF
7 pages

Loading next page...
 
/lp/wiley/a-stopped-flow-rapid-freezing-machine-with-millisecond-time-resolution-CYPjgHCSgY

References (11)

Publisher
Wiley
Copyright
Copyright © 1990 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1059-910X
eISSN
1097-0029
DOI
10.1002/jemt.1060160206
pmid
2213238
Publisher site
See Article on Publisher Site

Abstract

We have developed an instrument capable of freezing transient intermediates in rapid biochemical reactions for subsequent freeze‐fracturing, replication, and viewing by transmission electron microscopy. The machine combines a rapid mixing unit similar to one widely used in chemical kinetics (Johnson, 1986) with a propane jet freezing unit previously used to prepare static samples for freeze‐fracturing (Gilkey and Staehelin, 1986). The key element in the system is a unique thin‐walled flow cell of copper that allows for injection and aging of the sample, followed by rapid freezing. During freeze‐fracturing, a tangential cut is made along the wall of the flow cell to expose the sample for etching and replication. The dead time required for mixing and injection of the reactants into the flow cell is less than 5 ms. Electronic controls allow one to specify, on a millisecond time scale, any time above 5 ms between initiation of the reaction and quenching by rapid freezing.

Journal

Microscopy Research and Technique – Wiley

Published: Oct 1, 1990

Keywords: ; ; ; ;

There are no references for this article.