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Make sure there is enough space between colonies for the ring so as to avoid contamination; otherwise, use smaller rings if available If using cells from small colonies (35 50 cells), the Eppendorf washing stage can be omitted and the cells with the trypsin/EDTA transferred directly into 12-well plates containing a minimum of 2 mL of growth medium, which is enough to neutralize the trypsin As with any extracellular matrix, that in cartilage becomes increasingly crosslinked with age Consequently, the digestion time will vary with age of the donor, and tissue from younger donors will require less time for full digestion It should be remembered that the minimum digestion time for cell release should be striven for in order to maximize cell viability Dissolution of the tissue can also be expedited by sharp agitation at hourly intervals when in collagenase.



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system is illustrated by the pixel response curves in Fig. 10.42(b). These curves were obtained by focusing a white point target on the coded focal plane. The output of adjacent pixels is plotted as the target is scanned across the column. The extent of the pixel response is somewhat greater than 5.2 mm because of the nite extent of the target. The period of the pixel response curves is 5.2 mm. Although the mask pattern was not precisely registered to the pixels in this experiment, subpixel modulation of the response is indicated by the twin lobe structure of the pixel response. We analyze pixel mask-based focal plane coding by modeling each detector as an n n block of subpixels. The output of the ith detector is gi X hij fj (10:53)





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As a guide, use a minimum of 7 mL of digestion medium per gram of cartilage weight, although excess volume also works but is wasteful of reagents In the case of very young donors, refer to the fetal protocol that follows When the clonal cell lines have been transferred to 6-well plates or 35-cm Petri dishes, and if the culture is growing well, that is, is con uent within a few days, then further passaging can involve splitting at a 1:2 dilution rate Greater dilutions (1:4 or 1:8) can be made with more mature cultures It is important when using collagenases that you work in units of activity since there can be variations of some threefold between batches Batch testing is also advised..

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When the capital projects fund spends resources to purchase or construct an asset it charges an expenditure on its operating statement There is no asset recorded on the balance sheet of the fund Similarly, do not expect to find a liability on the balance sheet of the capital projects fund for the long-term debt issued to provide funds to purchase or construct the capital assets You will find the long-term liability for the debt only on the government-wide statement of net assets When proceeds from long-term debt are received by the capital projects fund, the operating statement records an other financing source of funds, which, although titled differently, works just like a revenue in the operating statement The astute reader may notice that long-term is used in the above discussion about whether the debt is recorded in the capital projects fund.

To view NAT statistics, select STATUS STATISTICS to display the Statistics screen. Scroll down to the Security area and click the NAT Stats button to display the NAT statistics.

A. May the Federal Government Require Manufacturers to Install EDRs Under the Motor Vehicle Safety Act, the DOT, on advice from the NHTSA, may promulgate, through informal agency rulemaking, federal highway safety standards, including manufacturer s safety component requirements. Both the Motor Vehicle Safety Act and the 1974 amendments concerning occupant crash protection standards indicate that motor vehicle safety standards are to be promulgated under the informal rulemaking procedures of the Administrative Procedure Act ( lPA )).23

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