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Cryo: Difference between revisions
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== Freezing in liquid Propane == | == Freezing in liquid Propane == | ||
It is proposed that freezing in liquid propane is faster and therefore better for a protein than in liquid nitrogen. Measurements with small thermocouples by Hakan Hope [http://www.bio.net/bionet/mm/xtal-log/1995-November/002005.html]revealed that there is almost no difference in cooling rates; cooling in liquid nitrogen was even faster than in liquid propane. On the other hand Teng and Moffat [http://scripts.iucr.org/cgi-bin/paper?wb0048] showed that flash cooling in liquid propane is fastest. My experience is that some but not all crystals freeze 'better' in liquid propane than in nitrogen. I frequently lost small crystals in crystallization buffers with low surface tension, e.g. high concentrations of MPD when plunged into liquid nitrogen. Freezing in liquid propane solved the problem. Also transport of crystals embedded in solid propane is easier than in liquid nitrogen: you can take your dry-shipper with you without any liquid nitrogen left in the | It is proposed that freezing in liquid propane is faster and therefore better for a protein than in liquid nitrogen. Measurements with small thermocouples by Hakan Hope [http://www.bio.net/bionet/mm/xtal-log/1995-November/002005.html]revealed that there is almost no difference in cooling rates; cooling in liquid nitrogen was even faster than in liquid propane. On the other hand Teng and Moffat [http://scripts.iucr.org/cgi-bin/paper?wb0048] showed that flash cooling in liquid propane is fastest. My experience is that some but not all crystals freeze 'better' in liquid propane than in nitrogen. I frequently lost small crystals in crystallization buffers with low surface tension, e.g. high concentrations of MPD when plunged into liquid nitrogen. Freezing in liquid propane solved the problem. Also transport of crystals embedded in solid propane is easier than in liquid nitrogen: you can take your dry-shipper with you without any liquid nitrogen left in the container. | ||
Here a short protocol to prepare liquid propane and how to use it. | Here a short protocol how to prepare liquid propane and how to use it. | ||
* Cryogenic burns are painful. I wear usually a combination of cotton gloves and latex gloves. | * Cryogenic burns are painful. I wear usually a combination of cotton gloves and latex gloves. | ||
* Use pure propane | * Use pure propane | ||
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* Put Pasteur pipette into the plastic tube with the metal piece. Propane will condensate on the cool metal piece. | * Put Pasteur pipette into the plastic tube with the metal piece. Propane will condensate on the cool metal piece. | ||
* After 0.5-1 min you should have 20-30 ml of propane. | * After 0.5-1 min you should have 20-30 ml of propane. | ||
* If you want to store the propane leave it in the liquid nitrogen until it is solid. | * If you want to store the propane leave it in the liquid nitrogen until it is solid. This might take a few minutes. | ||
* I fill the liquid propane into vials. For short time storage and crystal handling I put the vials in a flat liquid nitrogen bath. If propane becomes solid after a while just remove the vial from the liquid nitrogen and put it on the desk; let is stand for a while and when it is liquid again put it pack into the nitrogen bath. | * I fill the liquid propane into vials. For short time storage and crystal handling I put the vials in a flat liquid nitrogen bath. If propane becomes solid after a while just remove the vial from the liquid nitrogen and put it on the desk; let is stand for a while and when it is liquid again put it pack into the nitrogen bath. | ||
* I plunge the crystals just into the vials and wait until the propane is solid. | * I plunge the crystals just into the vials and wait until the propane is solid. | ||