I´m trying to prepare sections from isolated mitochondria, but the sections are very fragile. I used -120°C, 60nm and nickel coated grids but without any results.
Hello,
Are your mitochondria embedded in gelatin for cutting?
Do you pick up the section with a mixture of methyl cellulose-sucrose?
Hi Nicolas,
thank you for your answer. I have got the mitochondia embedded in the gelatine and as pick up solution I use the mixture of methyl cellulose- sucrose. I don´t know where I have problem...wrong infiltartion, temperature or grid??
Hi Romana,
If the sections are fragile but somewhat shiny and without compression, you could try lowering the sectioning temperature to -110 C or even -100 C. The 'higher' temperature will make the blocks a bit softer and the sections less fragile. You might have to change the knife position more often though as remnants of the section will stick more to the knife edge.
Cheers,
Rob
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