Şeffaflaştırılmış doku görüntülemesi için ışık tabakası floresans mikroskobu geliştirilmesi

Light sheet fluorescence microscopy has become a popular technique for life sciences such as biomedical studies due to its advantages compared to confocal and epifluorescence microscopy. LSFM is a non-destructive technique using a narrow line of light; instead of a focused spot, for the illumination and it allows fast imaging with large field of view and reduced photobleaching effect. In this study, we present development of a home-built LSFM for cleared tissue imaging set-up with commercially available equipment. LSFM involves formation of a thin sheet of laser light using a cylindrical lens. The sample is scanned along the light sheet; thus the sample planes are selectively illuminated. The selective illumination of the sample allows for selective fluorescent excitation at desired focal planes. Besides three-dimensional image of the sample is acquired by the reconstruction of selected planes. The LSFM has been built utilizing Open Access platform for Selective Plane Illumination Microscopy. The sample was a stained cleared tissue with fluorescent tagged secondary antibodies. In order to illuminate the sample, several laser sources with a wavelength of 525 nm was used for example a Coherent Chameleon tunable laser with an SHG and a diode laser. Three-dimensional images were reconstructed from the imaged planes using the Fiji ImageJ software.