Optical (Light) Microscope Eye Final Image- Eyepiece Projecto Objective specim en Condenser Lens Light 16
16 Optical (Light) Microscope
lumination Source petra of common Lamps 4000 Laser Spectrum Incandescent 1500 1000 kah Wavelength(nm) hore laser Ho: YAG Argon Er: Fiber CrEr: YSSG Alexandrite Nd: YAG Er: YAG co2 Diode 10a00b 10000 20000 Wavelength(nm) 17
17 Illumination Source Laser Spectrum
Numerical Aperture(N.A,数值孔径) Aperture Numerical Aperture and Airy Disc Size (x-y'Hp Objoct 沙國!罗 Entrance Figure 4 N.A. =n sina · The higher na. of the 增大透镜经 objective lens, the better resolution Methods to improve the 1.5 N.A. of an objective lens 缩短物镜焦距 增大介质的折射率 18
18 Numerical Aperture（N. A., 数值孔径） • N.A. = n sinα • The higher N.A. of the objective lens, the better resolution. • Methods to improve the N.A. of an objective lens
Bright Field and Dark Field Imaging B objective pring Ins diaphragm dark-field diffracted (deviated) light object” B Dark field can improve contrast not resolution 19
19 Bright Field and Dark Field Imaging • Dark field can improve Contrast, not resolution!
Confocal Fluorescence Microscope Detector Laser Scanning Confo Optical out→ of-Focus Configuration Filter Excitation Lase Light Rays Dichromatic xcitation Objective rce Focal fluorescent specimen is illuminated emitted flu emitted light from out- Planes Figure 2 a pinhole point is for focus at pinhole and is largely excluded Confocal and Widefield Fluorescence Microscopy detector from detector Figure 1 (f)
20 Confocal Fluorescence Microscope