Filters emission

In this case, the excitation and emission filters and dichroic mirror used with the sample are removed and replaced with a beam splitter [3, 36], A scattering solution is placed on the microscope and a... 

The accuracy with which a system can measure lifetimes depends on a number of different factors including calibration of the instrument, the number of detected photons and also the efficiency of the analysis routines. In addition, sources of background and scattered light should be eliminated. Emission filters should be chosen with great care to make sure that no scattered laser light reaches the detector. Detection of scattered excitation light results in a spurious fast component in the decay and complicates the interpretation of the data. The choice of emission filters is much more critical in FLIM than in conventional fluorescence intensity imaging methods. 

D Don Don raw donor image collected at with donor emission filter (donor channel)... 

S Don s.e. raw sensitized emission image collected at with the sensitized emission filter (s.e. channel)... 

A Acc Acc raw acceptor image collected at l x with the acceptor emission filter (acceptor channel)... 

The fluorescent components are denoted by / (intensity) followed by a capitalized subscript (D A or s, for respectively Donors, Acceptors, or s.e.) to indicate the particular population of molecules responsible for emission and a lower-case superscript ( " or ) that indicates the detection channel (or filter cube). For example, I denotes the intensity of the donors as detected in the donor channel and reads as Intensity of donors in the donor channel, etc. Notes (1) The excitation in the s.e. channel is generally set up to be equal to that in the donor channel. In case a separate filter cube is used, slight differences may occur, which is denoted by Don(S). See the text and appendix for further details. (2) The s.e. emission filter is usually the same as the acceptor emission filter in confocal determinations. We here designate a different filter to accommodate those wide-field/digital camera experiments that employ different filters for A and S. (3) Here the notation D-S indicates the residual (quenched) donor fluorescence in the presence of the acceptor. In the other chapters this is indicated as DA. Hence ... 

A final distinction is that on confocal microscopes S and A images are commonly acquired with the exact same emission filter settings whereas for CCD microscopes they typically involve physically separate- and therefore slightly different—filter cubes.6 This simplifies the calculation of leak-through terms [3], In Appendix of this chapter, we rather generalized the treatment of filterFRET by not making assumptions on the filter settings for S and A. 

Relates signal from cross-excited donors in S to that in A (provided that emission filters are identical)... 

Fig. 10.2. FSPIM analysis of the interaction between maize transcriptional coactivators—GCN5 and ADA2—fused to CFP and YFP. GCN5 is a histone acetyltransferase that, in conjunction with adaptor protein ADA2, modulates transcription in diverse eukaryotes by affecting the acetylation status of the core histones in nucleosomes [63]. CFP- and YFP-tagged proteins, expressed in protoplasts, were excited by the 458 nm and the 514 nm laser lines sequentially. CFP fluorescence was selectively detected by an FIFT 458 dichroic mirror and BP 470-500 band pass emission filter while YFP fluorescence was selectively detected by using an HFT 514 dichroic mirror and...

Signals for methyl paraben were monitored with UV detection at 254 nm. The signal for rhodamine 110 chloride was monitored via fluorescence detection with an excitation filter of 482 nm (35 nm bandwidth) and emission filter of 535 nm (40 nm bandwidth). A gradient method (same as the one in Figure 6.16) was used. The compositions of mobile phases A and B were 5 95 H20 CH3CN with 0.1 HCOOH and CH3CN with 0.085% HCOOH, respectively, with a total flow rate of 300 fiL/ min (corresponding to 12.5 /rL/min for each column). 

Fluorophores can be visualized in fluorescence microscopy using special filter blocks that are composed of the excitation filter, dichroic mirror and emission filter. The excitation filter must select wavelengths of light from a light source that fall in the maximum absorption region of the fluorophore. The emission filter must pass the fluorescent wavelengths but not the excitation wavelengths. The dichroic mirror... 

Emission filter must pass the fluorescent wavelengths but not the excitation wavelengths. 

The detection performance of an LIF photometric device is governed by the emission filter(s), excitation filter(s), detector type, the excitation source and the detection scheme. The selection of optical elements and device configuration as it relates to the detection performance is further described by expanding the collection efficiency term in Equation 11.3 ... 

Dust (especially from industrial activities) and salt spray will also exacerbate atmospheric corrosion (Section 16.4). In enclosed industrial premises, atmospheric corrosion could be minimized by preventing noxious emissions, filtering the air to remove particulate matter, and scrubbing the air with water to remove SO2 and other objectionable gases, although the humidity should itself be kept as low as possible (e.g., steam leaks should not be tolerated). On the global scale, however, the cost to the public of atmospheric corrosion could be substantially reduced by sharply limiting SO2 and, to a lesser extent, NO. emissions from power plants, smelters, automobiles, and other industrial functions. This is an aspect of the acid rain threat (Chapter 8) that is usually overlooked. 

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