研究目的
Comparing the depth discrimination and speed performance of multifoci Raman hyperspectral imaging with the reference standard of a single laser point confocal Raman mapping.
研究成果
The multifoci excitation approach provides a clear speed advantage while maintaining a high depth discrimination. An R = 8 period was optimal for maintaining depth discrimination while maximizing imaging speed.
研究不足
The practical number of laser foci is limited by the small field of view of the high numerical aperture (NA) objectives used in confocal Raman microscopy. The overlapping laser light corresponding to different foci leads to cross‐talk between the Raman spectra detected on the CCD and thus degrades the depth discrimination.
1：Experimental Design and Method Selection:
A liquid crystal spatial light modulator (LC‐SLM) was employed for the generation of multifoci laser beams, and a digital micromirror device (DMD) was used as a software‐configurable reflective pinhole array. The patterns of the laser foci and pinhole array can be rapidly changed without requiring any hardware alterations. Confocal patterns with different distance‐to‐size ratios were tested and compared.
2：Sample Selection and Data Sources:
Samples included a uniform polystyrene slide, a thick transparent sample (oil), stacked diphenylalanine (FF) microtubes, and Acanthamoeba castellanii trophozoites.
3：List of Experimental Equipment and Materials:
LC‐SLM, DMD, CW Ti:Sapphire laser, 60×/1.2 NA water‐immersion objective, spectrometer.
4：2 NA water‐immersion objective, spectrometer.
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Raman spectra were acquired at different focal planes by moving the sample in the axial direction (z‐axis) in 10‐μm steps. The depth response curves were obtained by integrating the Raman bands corresponding to oil.
5：Data Analysis Methods:
The first derivatives of the response curves were fitted with a Gaussian function to calculate the full width at half maximum (FWHM).
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容
