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In this section, we discuss some interesting properties of the considered algorithms, and present also an alternative derivation of the robust ICA algorithm proposed by Cruces et al. [321, 332]. Only time-discrete learning algorithms are considered. The learning algorithm (7.25) can be derived using the basic concepts of the diagonalization of a generalized covariance matrix: Rf g (W) = E{f (y)gT (y)}. (7.36)

= 2,



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antibiotics as soon as possible If the culture grows well, then antibiotics can be removed from the bulk of the stocks at rst subculture, retaining one culture in antibiotics as a precaution if necessary Antibiotics can lead to lax aseptic technique, can inhibit some eukaryotic cellular processes, and can hide the presence of a microbial contamination If a culture is contaminated this must become apparent as soon as possible, either to indicate that the culture should be discarded before it can spread the contamination to other cultures or to indicate that decontamination should be attempted The latter should only be used as a last resort; decontamination is not always successful and can lead to the development of antibiotic-resistant organisms Most bacterial, fungal, and yeast infections are readily detected by regular careful examination with the naked eye (eg.





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2 4 [S4 (y)]jj = sign E{yj } 3(E{yj })2 = sign 4 (yj ).

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, by a change in the color of culture medium) and on the microscope However, one of the most serious contaminations is mycoplasma, which is not visible by routine microscopy Any cell culture laboratory should have a mycoplasma screening program in operation, but those collecting tissue for primary culture are particularly at risk The precautions that should be observed are as follows: (i) Treat any new material entering the laboratory from donors or from other laboratories as potentially infected and keep it in quarantine Ideally, a separate room should be set aside for receiving samples and imported cultures If this is not practicable, handle separately from other cultures, preferably last in the day and in a designated hood, and swab the hood down after use with 2% phenolic disinfectant in 70% alcohol Use a separate incubator, and adhere strictly to the rules given above regarding medium sharing.

Figure 2.2 Example of a basis function zit of the space V,,

Screen new cultures as they arrive, and existing stocks at regular intervals, eg, once a month There are a number of tests available, but the most reliable and sensitive are uorescence microscopy after staining with Hoechst 33258 [Chen et al, 1977; Protocol 192, Freshney, 2005] or PCR with a panmycoplasma primer [Uphoff and Drexler, 2002a; Protocol 193, Freshney, 2005] The latter is more sensitive but depends on the availability of PCR technology, whereas the former is easier, will detect any DNA-containing contamination, but requires a uorescence microscope Both techniques are best performed with so-called indicator cultures The test culture is refreshed with antibiotic-free medium and, after 3 5 days, the medium is transferred to an antibiotic-free, 10% con uent indicator culture of a cell line such as 3T6 or A549 cells, which are well spread and known to support mycoplasma growth.

FIGURE 3.44 After adding text-based notes or audio comments to an image, save the file in PDF so that others can access the annotations when viewing the image in Adobe Reader.

Now that you understand how the curve works, we discuss the features of the Curve tool that enable you to create dynamic curves that can really have an impact on the tones of your images. These tools can be found whether you are adjusting images using the Curves tool, with an adjustment layer, or in Camera Raw.

After a further 3 days (the indicator cells must not be allowed to reach con uence) the indicator culture is xed in acetic methanol and stained with Hoechst 33258, or harvested by scraping for PCR (trypsinization may remove the mycoplasma from the cell surface) Discard all contaminated cultures If the culture is irreplaceable, decontamination may be attempted (under strict quarantine conditions) with agents.

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