Limits...
Some personal and historical notes on the utility of "deep-etch" electron microscopy for making cell structure/function correlations.

Heuser JE - Mol. Biol. Cell (2014)

Bottom Line: Then it explains our approach to freezing, namely the Van Harreveld trick of "slamming" living cells onto a supercold block of metal sprayed with liquid helium at -269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment.This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most "true to life" views of cells out of them with a minimum of hassle.Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the "founding fathers" that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are "irresistible and half transparent … their meaning buried under only a few years of work," and "reasonable working hypotheses are already suggested by the ultrastructural organization itself."

View Article: PubMed Central - PubMed

Affiliation: WPI Institute, Kyoto University, Kyoto 606-8501, Japan; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 jheuser@icems.kyoto-u.ac.jp jheuser@wustl.edu.

Show MeSH
John E. Heuser
© Copyright Policy - creative-commons
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4214773&req=5

Figure 1: John E. Heuser

Mentions: Deep-etch EM is a variant of what Moor introduced (Heuser and Salpeter, 1979) and deserves special attention only because its purpose has been to avoid all of the fixation and staining and dehydrating procedures that had accompanied previous approaches to EM and essentially to get living cells replicated after they were frozen (Figure 1). We found that freeze fracture works just as well or better on unfixed cells and molecules, and therefore made a huge effort to devise a really good way to freeze living cells, tissues, and cell extracts without introducing such artifacts as ice-crystal damage.


Some personal and historical notes on the utility of "deep-etch" electron microscopy for making cell structure/function correlations.

Heuser JE - Mol. Biol. Cell (2014)

John E. Heuser
© Copyright Policy - creative-commons
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4214773&req=5

Figure 1: John E. Heuser
Mentions: Deep-etch EM is a variant of what Moor introduced (Heuser and Salpeter, 1979) and deserves special attention only because its purpose has been to avoid all of the fixation and staining and dehydrating procedures that had accompanied previous approaches to EM and essentially to get living cells replicated after they were frozen (Figure 1). We found that freeze fracture works just as well or better on unfixed cells and molecules, and therefore made a huge effort to devise a really good way to freeze living cells, tissues, and cell extracts without introducing such artifacts as ice-crystal damage.

Bottom Line: Then it explains our approach to freezing, namely the Van Harreveld trick of "slamming" living cells onto a supercold block of metal sprayed with liquid helium at -269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment.This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most "true to life" views of cells out of them with a minimum of hassle.Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the "founding fathers" that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are "irresistible and half transparent … their meaning buried under only a few years of work," and "reasonable working hypotheses are already suggested by the ultrastructural organization itself."

View Article: PubMed Central - PubMed

Affiliation: WPI Institute, Kyoto University, Kyoto 606-8501, Japan; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 jheuser@icems.kyoto-u.ac.jp jheuser@wustl.edu.

Show MeSH