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triplet state typically has only slightly higher energy than the lowest-energy singlet state A third phenomenon resulting in radiative transitions called chemiluminescence occurs in molecules which can be promoted to an excited state as a result of a chemical reaction and which then return to the ground state with the emission of light Molecules such as luciferin and luminol (Table 34) show this property and, when used as substrates by the enzymes re y luciferase and peroxidase, respectively, emit light which may be measured or detected (Section 342) These reactions are highly ef cient and may be readily incorporated into sensitive assay systems or in staining of protein or nucleic acid blots (Figure 322).



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For example the level of ATP (or of any biomolecule which may be coupled to ATP production) may be quanti ed as follows: Luciferin + ATP + O2 (Fire y Luciferase) Oxyluciferin + AMP + pyrophosphate + CO2 + light This assay is extremely sensitive allowing detection at the pM/fM level Since peroxidase-conjugated antibodies are extensively used in immunodetection systems, luminol in particular has wide applications in this general area (eg Section 5102) Both phosphorescence and chemiluminescence are related to uorescence in the sense that all three phenomena result in emission of light Fluors and molecules capable of chemiluminescence are often found to possess complex ring structures which are largely responsible for their rigidity (Table 34) Most components of biopolymers are non uorescent and this means that uorescence may give information about processes occurring in speci c parts of the biopolymer.





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For example, the most important intrinsic uors in proteins are tryptophan, tyrosine and phenylalanine residues (in practice, tryptophan and tyrosine give stronger spectra than phenylalanine) Fluorescence of tyrosine is frequently quenched as a result of proton transfer in the excited state (see below) These residues have average occurrences of only 35, 11 and 35% in proteins, respectively Accordingly, many proteins with extensive absorbance spectra may have only one or two uorescent residues and advantage can be taken of this in the study of biochemical processes involving the protein The bases of DNA nucleotides and of some enzyme cofactors (eg NAD) are also intrinsic uors although they display generally weak uorescence spectra In such cases, extrinsic uors may be used to carry out structural and functional studies These are nonbiological molecules which possess uorescence properties (Table 34)..

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Thaw/Freeze: You can freeze layers for reasons similar to those that lead you to turn off layers. In general, freeze layers when you want to work without those layers for a longer period of time. The thawed/frozen state is available on the Layer Control dropdown list. Thaw/Freeze in Current Viewport: You may want to freeze or thaw layers in some floating viewports but not in others. For example, if you want to display dimensions in one viewport but not in others, freeze your dimensions layer in all of the viewports except one. You can change this state on the Layer Control drop-down list if you re working on a layout. You can also click the Current Viewport Freeze icon in the Layer Properties Manager. Thaw/Freeze in New Viewports: You can also freeze or thaw layers in future viewports that you create. For example, after you display dimensions or text in one viewport, you may want to make sure that they won t appear in any new viewports that you create. This state is available in the New Viewport Freeze column of the Layer Properties Manager.

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2. In AutoCAD, type (load file name ) where file name is the name of the LSP file. You don t need the .lsp, but don t forget the parentheses or the quotation marks. Alternatively, choose Tools Load Application to use a dialog box. AutoCAD responds with the name of the last function that was defined in the routine. 3. Type the name of the function to use the AutoLISP routine. Most of the AutoLISP routines were written before AutoCAD 2007 came out. Sometimes the installation instructions are out of date and don t include the capability of adding the folder to the support file search path. If no text file explains how to use the program, brief instructions may be displayed on the command line. If not, type the name of the file, such as atc, and press Enter.

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