利用流的式细胞仪测量拉曼光谱 xin.pdf

Original Article A Flow Cytometer for the Measurement of Raman Spectra Dakota A. Watson,1 Leif O. Brown,2 Daniel F. Gaskill,1 Mark Naivar,2 Steven W. Graves,2 Stephen K. Doorn,2 John P. Nolan1* Abstract 1La Jolla Bioengineering Institute, Multiparameter measurements in flow cytometry are limited by the broad emission La Jolla, California 92037 spectra of fluorescent labels. By contrast, Raman spectra are notable for their narrow 2 Los Alamos National Laboratory, spectral features. To increase the multiparameter analysis capabilities of flow cytometry, Los Alamos, New Mexico 97545 we investigated the possibility of measuring Raman signals in a flow cytometry-based system. We constructed a Raman Spectral Flow Cytometer, substituting a spectrograph Received 14 September 2007; Accepted and CCD detector for the traditional mirrors, optical filters, and photomultiplier tubes. 27 November 2007 Excitation at 633 nm was provided by a HeNe laser, and forward-angle light scatter is Grant sponsor: NIH; Grant numbers: used to trigger acquisition of complete spectra from individual particles. Microspheres R01 EB003824, P41 RR01315, were labeled with nanoparticle surface enhanced Raman scattering (SERS) tags and R21 RR020064, LANL LDRD. measured using the RSFC. Fluorescence and Raman spectra from labeled microspheres were acquired using the Raman Spectral Flow Cytometer. SERS spectral intensities were *Correspondence to: John P. Nolan, 505