howto:how_to_measure_the_instrument_response_function_irf
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howto:how_to_measure_the_instrument_response_function_irf [2016/12/06 11:50] – veiga | howto:how_to_measure_the_instrument_response_function_irf [2016/12/06 12:05] – veiga | ||
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Put a droplet on a coverslip, measurement conditions as for fluorescence measurement | Put a droplet on a coverslip, measurement conditions as for fluorescence measurement | ||
</ | </ | ||
- | ((Szabelski M., Iliev D., Sarkar P., Luchowski R., Gryczynski Z., Kapusta P., Erdmann R., Gryczinski I.\\ | + | |
- | Collisional quenching of Erythrosine B as a potential reference dye for impulse response function evaluation\\ | + | |
- | Applied Spectroscopy, Vol.63, p.0363-0368 (2009)\\ | + | ==== Two photon excitation |
- | http://www.ingentaconnect.com/ | + | |
+ | Do not attempt to record an [[glossary: | ||
+ | |||
+ | You can try to excite (by [[glossary: | ||
+ | |||
+ | With microscopes it is convenient to record the second harmonic signal that is generated on the surface of urea crystals. The best is to let evaporate a droplet of concentrated urea solution on a clear cover slip. The resulting film of micro-crystals is easy to target. | ||
+ | |||
+ | Urea, aka Carbamide or Carbonyldiamide, | ||
+ | |||
+ | ===== Appropriate Count Rate for Measuring an IRF ===== | ||
+ | See [[glossary: | ||
+ | |||
+ | ===== How to compensate IRF effects in the analysis of time domain measurements ===== | ||
+ | |||
+ | There are two major ways of compensating IRF effects: | ||
+ | |||
+ | *correct the effects in the data (**de**convolution) | ||
+ | *take the effects into account in your model equation (**re**convolution) | ||
+ | |||
+ | Note: All analysis packages from PicoQuant use the [[glossary: | ||
+ | |||
+ | ===== Measuring the IRF as scattered excitation light ===== | ||
+ | |||
+ | We do not recommend to measure the IRF as scatters light in microscopy, due to the color dependence of SPAD detectors, which are generally used in microscopy. Furthermore, | ||
+ | |||
+ | However, in case of cuvette based measurement like in spectrometers, | ||
+ | |||
+ | Note that recording the IRF via scattering requires tuning the emission monochromator to the excitation wavelength. In filter based machines, e.g. FluoTime100 this means removing the emission bandpass or longpass filter. In microscopes, | ||
- | ==== Selected literature | + | ==== Selected literature: ==== |
Luchowski R., Kapusta P., Szabelski M., Sarkar P., Borejdo J., Gryczynski Z., Gryczynski I.\\ | Luchowski R., Kapusta P., Szabelski M., Sarkar P., Borejdo J., Gryczynski Z., Gryczynski I.\\ | ||
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- | ==== Two photon excitation (TPE) ==== | ||
- | |||
- | Do not attempt to record an [[glossary: | ||
- | |||
- | You can try to excite (by [[glossary: | ||
- | |||
- | With microscopes it is convenient to record the second harmonic signal that is generated on the surface of urea crystals. The best is to let evaporate a droplet of concentrated urea solution on a clear cover slip. The resulting film of micro-crystals is easy to target. | ||
- | |||
- | Urea, aka Carbamide or Carbonyldiamide, | ||
- | |||
- | ===== Appropriate Count Rate for Measuring an IRF ===== | ||
- | See [[glossary: | ||
- | |||
- | ===== How to compensate IRF effects in the analysis of time domain measurements ===== | ||
- | |||
- | There are two major ways of compensating IRF effects: | ||
- | |||
- | *correct the effects in the data (**de**convolution) | ||
- | *take the effects into account in your model equation (**re**convolution) | ||
- | |||
- | Note: All analysis packages from PicoQuant use the [[glossary: | ||
- | |||
- | ===== Measuring the IRF as scattered excitation light ===== | ||
- | |||
- | We do not recommend to measure the IRF as scatters light in microscopy, due to the color dependence of SPAD detectors, which are generally used in microscopy. Furthermore, | ||
- | |||
- | However, in case of cuvette based measurement like in spectrometers, | ||
- | |||
- | Note that recording the IRF via scattering requires tuning the emission monochromator to the excitation wavelength. In filter based machines, e.g. FluoTime100 this means removing the emission bandpass or longpass filter. In microscopes, | ||
howto/how_to_measure_the_instrument_response_function_irf.txt · Last modified: 2023/09/07 22:55 by peter