Download PDF by BSI: BS EN 1396:1997 Aluminium and aluminium alloys - Coil coated

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ISBN-10: 0580271447

ISBN-13: 9780580271441

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Extra info for BS EN 1396:1997 Aluminium and aluminium alloys - Coil coated sheet and strip for general applications - Specifications

Example text

5. The nuclear fraction (8008, 10 mm) 1s washed only once. Supernatant fluid 1s then centrifuged at 20,OOOg for 20 mm to yield the mitochondrlal (P2) fraction The P2 fraction is resuspended m water (2 mL/g tissue) and sublected to density gradient centrifugation, as described by Whittaker et al. 4M sucrose contammg the mhibitors placed at the bottom of the density gradient, which 1s then centrifuged at 93,OOOg for 2 h. 8M sucrose) Electron microscopy of subfractions prepared by density gradient centnfugatlon of shocked P2 fractions appeared essentially as detailed by Whittaker (1969), except for subfractions D and E.

Micropunch From Microwave-Irradiated Brains To avoid postmortem changes, the heads of the animals are irradiated by focused microwaves. These brains need special care, They are difficult to remove from the skull because they break into pieces easily. They need to be sectioned in a cryostat but at a lower temperature (-15’C) than that used for fresh brains. It is helpful to cool the knife blade intermittently with dry ice. Topographical orientation is quite difficult since the “cooked” brain is almost equally grayish.

The temperature of the sections is of malor importance. If they are too cold, they are prone to break and if they are too warm, they are melted beneath the needle. The dissected tissue pellet can be blown out of the needle, or pushed out with a stylet mto a tube or dish, or onto the tip of a microhomogenizer. If several samples are punched from the same bram nucleus, the pellets need not be blown out separately-they may be collected in the lumen of the needle and removed all at once After micropunch, sharp-edged holes remam m the section (Figs.

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