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Advanced Techniques for Immuno-Labelling and Sample Processing for TEM

This guide outlines the step-by-step procedures for immuno-labelling and sample processing tailored for transmission electron microscopy (TEM). Key procedures detailed include cell/tissue fixation with 4% PFA, washing and quenching processes, gelatin embedding with 2.3M sucrose infusion, and cryo-sectioning. Additionally, it covers trimming, section pick-up, EM grid application, and contrasting for enhanced imaging. Each step is designed to optimize the sample's quality and the accuracy of results. For detailed protocols, visit our comprehensive resource.

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Advanced Techniques for Immuno-Labelling and Sample Processing for TEM

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  1. liquid nitrogen cryo-chamber temp. control sectioning control Immuno-labelling for TEM Sample processing Fix 4% PFA Wash/Quench Harvest cells/tissue 3a. Gelatin embedding 2.3M sucrose infusion Cut ≤1mm3 pieces Mount on cryo-stub Snap freeze liquid N2 http://www.liv.ac.uk/emunit

  2. 1. 2. 3. 4. Immuno-labelling for TEM Trim Section Pick-up Add to EM grids Grid looping Drain excess Dry Immuno-labelling MC/UA contrasting http://www.liv.ac.uk/emunit

  3. Block Wash Quench 1°Ab 5. 6. Immuno-labelling for TEM 50µl washes 5µl/grid Ab incubations Trim Section Pick-up Add to EM grids Immuno-labelling MC/UA contrasting Grid looping Drain excess Dry http://www.liv.ac.uk/emunit

  4. 7. 8. 9. Immuno-labelling for TEM Trim Section Pick-up Add to EM grids Immuno-labelling MC/UA contrasting Grid looping Drain excess Dry http://www.liv.ac.uk/emunit

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