ABSTRACT
This chapter discusses the best to apply fluorescence lifetime microscopy (FLIM) to those autofluorescent cellular components and analyzes the data by individual, segmented cells using HeLa cervical cancer cells as a model. nicotinamide adenine dinucleotide and flavin adenine dinucleotide channels also contain 690 nm short pass filter in the beam path. Evolution has provided life on earth with several methods of generating cellular energy. Applications of FLIM have grown exponentially in a broad range of life sciences and industrial fields, a reflection of specific advantages over intensity-based microscopy. The effects on lifetime of fixing specimens with formaldehyde or methanol have been published with mixed results for fluorescently labeled proteins, depending on types of buffers and/or mounting media. The fluorescence lifetime parameters are derived from a model containing lifetime components and fractions used to fit the measured data. The measured decay data are fitted with the convolution of the model function and the instrument response function.
