Label-Free Multiphoton Microscopy: Much More Than Fancy Images

International Journal of Molecular Sciences

KurtI. Anderson¹  Deborah Sandrin²  Andrea Filippi²  Filippo Romanato³  Giulia Borile³ 

[1]Crick Advanced Light Microscopy Facility (CALM), The Francis Crick Institute, London NW1 1AT, UK

[2]Department of Physics and Astronomy “G. Galilei”, University of Padua, 35131 Padua, Italy

[3]Laboratory of Optics and Bioimaging, Institute of Pediatric Research Città della Speranza, 35127 Padua, Italy

关键词: multiphoton microscopy;    label-free;    second harmonic generation;    third harmonic generation;    quantitative imaging;   

DOI  :  10.3390/ijms22052657

来源: DOAJ

【 摘 要 】

Multiphoton microscopy has recently passed the milestone of its first 30 years of activity in biomedical research. The growing interest around this approach has led to a variety of applications from basic research to clinical practice. Moreover, this technique offers the advantage of label-free multiphoton imaging to analyze samples without staining processes and the need for a dedicated system. Here, we review the state of the art of label-free techniques; then, we focus on two-photon autofluorescence as well as second and third harmonic generation, describing physical and technical characteristics. We summarize some successful applications to a plethora of biomedical research fields and samples, underlying the versatility of this technique. A paragraph is dedicated to an overview of sample preparation, which is a crucial step in every microscopy experiment. Afterwards, we provide a detailed review analysis of the main quantitative methods to extract important information and parameters from acquired images using second harmonic generation. Lastly, we discuss advantages, limitations, and future perspectives in label-free multiphoton microscopy.

【 授权许可】

Unknown