Movies Demonstrating Freeze-Grinding Techniques

The raw video for these movies was filmed by Dr. Robert Givens and me in late 2007 and early 2008, but the video wasn't converted by me into finalized movies until August, 2012. Dr. Givens and I hope that these movies will prove useful to the yeast research community.

The movies demonstrate some of the techniques described verbally in our 2011 publication (RM Givens, LD Mesner, JL Hamlin, MJ Buck and JA Huberman, Integrity of chromatin and replicating DNA in nuclei released from fission yeast by semi-automated grinding in liquid nitrogen. BMC Research Notes 2011, 4:499). The publication is available for free at Although these techniques were developed for fission yeast cells, they should be applicable with only minor modifications to budding yeast cells and to any other organism having tough cell walls.

Each of the movies is available from YouTube (see links below), and each is also available in .m4v format as a download from this website (see links below). The .m4v format will be useful to people who may wish to view the movie from a location not having Internet access. Note that the .m4v files are available in two forms: (1) native .m4v, and (2) compressed (zipped) .m4v. The compressed files are only slightly smaller than the non-compressed files. All are large and while require minutes to hours to download, depending on the speed of your Internet connection.

  1. "How to Freeze-Grind Fission Yeast Cells". This 42-minute movie describes in detail the process of grinding fission yeast cells in a precision-controlled motorized mortar-and-pestle grinder (Retsch RM100 or similar model) under liquid nitrogen. When carried out according to the instructions given here, intact nuclei are released in excellent yield from the broken cells.
  2. "Fine Freeze-Grinding of INCA Buffer to Promote its Uniform Mixing with Freeze-Ground Fission Yeast Cells". This 20-minute movie describes the process of making an ultra-fine frozen powder of INCA buffer, which can be mixed as a powder with a powder of frozen, ground fission yeast cells to generate a "cryo-mix". When the cryo-mix is brought to 0 degrees C or above, each yeast cell organelle and/or fragment will be immersed in properly mixed protective buffer as it melts.
  3. "Isolation of High Molecular Weight DNA from Freeze-Ground Fission Yeast Cells". This 18-minute movie describes the initial steps in the process of purifying high-molecular-weight DNA containing replication intermediates from a suspension of intact fission yeast nuclei (generated by freeze-grinding fission yeast cells followed by differential centrifugation).

If you have questions about, or problems with, any of these movies, please send me an e-mail (huberman AT buffalo DOT edu).