One-photon versus Two-photon Laser Scanning Mic roscopy and Digital Image Analysis of Microbial Biofilms

摘要: Publisher Summary Laser scanning microscopy (LSM) represents a standard optical technique in life sciences. LSM can be employed many different modes including reflection and fluorescence, single multichannel, one-photon two-photon, with data collection via intensity lifetime imaging. This chapter focuses on two-photon imaging of microbiological samples digital-image analysis the resulting sets. A structured approach to stepwise examine new, unknown biofilm sample has been outlined chapter. In this approach, advantage is taken intrinsic properties such as autofluorescence. Simple but specific staining for cellular polymeric constituents their distribution applied probes cell identity, viability well enzyme activity are discussed. The describes various microenvironment exception Most fluorochromes used excitation although excitation/emission response will from that (1P-LSM). Another option (2P-LSM) several may combined excited one wavelength only emission signals individually collected separate channels.