用MSV-5000系列紫外-可见光显微镜测量Volvox藻类的透射

MSV-5000系列显微分光光度计适用于从紫外到近红外的宽波长范围内的透射和反射测量。内置的高分辨率相机可以精确测量直径小至10μm的样本区域，因此在生物科学领域分析活细胞中的局部成分越来越受欢迎。团藻是一种藻类的形式，它在母体菌落中形成多达50,000个细胞的局部球形菌落。在本应用说明中，测量了团藻的子菌落，以获得吸收光谱和固定波长映射。Application NoteUV-0024 Transmission Measurement of Volvox using aMSV-5000 Series Microscopic Spectrophotometer2/3 Intro d uc t ion The MSV-5000 s e ries microscopic spec t rophotom e t e r i s for tr ansmi s sion and r e fl e ct i on measurements in a wide wav e le n gth r ange fr om the u ltr av i olet to nea r infrared . The bu i lt -in hig h r e sol ut ion camera enables sample ar e as a s smal l as 10um i n d i ameter to be prec i sely measured and i s there f ore gaining popu l ar i ty in the bioscience fi eld for analys in g l ocal i zed co n st i tuents in l i ving cel l s. Volvox is a f or m of algae whi c h forms locali ze d， spherical colon ie s of u p to 50，000ce l ls in th e mother colony. In t his application not e， a daught e r colony of Volvox was measured t o obtain the absorption spectra and fixed-wavele n gth mapping. Fig u re 1. Drie d v o lv o x . Keyword s MSV -5100 U V -V i s mi c ros c opi c s p ectro p hot o met e r ， MAXY-501 A u t o mati c XYZ s t ag e， Bioc h em i stry MSV-5200UV-Vi s i b le /NIR Microscopic Spectrophotom e ter 28600 Ma r y 's C ou r t， Easton， M D 21601 US A Appl i c a t i on Note Exper i men t al Absorption Fixed Wavelength Mapping UV-Vis Bandwidth 5nm Mode/Wavelength Lattice/672 nm Scan Speed 1000nm/min Bandwidth 2nm Response Quick Response Fast Data Interval 1nm Data interval 30 um Cassegrain Objective 16x Cassegrain objective 16x Aperture 50 mm中 Aperture 30 um中 R esults The measu r ed abso r ption sp e c t rum of Volvox i s shown in F i gure 2 on th e l eft . On t h e r i ght th e a b sorpt i on spectra of ch l oro p hyll a and b a re shown， which a r e major compone n ts of a l g a e and t h erefor e Volvox. T h e publis h ed lit erat u r e data i s measured u nd e r aceto n e solvent co n ditions a n d de p e n di n g on t h e solve n t condit i ons， t h e p eak posi ti ons o f ch l orophylls can be shi f t e d 2-7 nm . Howeve r， wh e n compar i n g the o b tai ne d Volvox sp e ct ru m to the l it e rat u r e c h loro p hyll s p ec t ra it can be ass u med tha t Vov l ox contains both c h lorophyll a a n d b. Figure 2. Absor p tion s pe ct r um o f Volvox (ri g h t) an d c hloro p hyll a an d b (r igh t). Fix e d waveleng th mapping measurements were o b ta i n ed a t 672 nm， the pea k observed i n t he absorpt i on spectrum ir F i gure 2. The hig h speed mapping i mages are shown below in F i gure 3. The areas with higher cellula r density are easily visuali ze d in both the sample obs e rvation image and color -cod e d diagram. 28600 Mary's Cour t ， E a s to n ， M D 21601US A UASGO 210 10 0 100 200 270 X [u m ] Figure 3. O b s e rva t ion i m age (lef t) a nd co l o r-co de d di a gram o f mappi ng m ea s ur em en t (r igh t) of mappin g . Re f e r en c e s 1. T anaka， Ryouichi a n d Ayumi Ta n aka.“Ch l oro p hyl l cycle regulates t h e construction and d e structio n of the light-harvesting complexes."Biochimica et Biophysica Acta (BBA)-Bioenergetics， 1807， 9， 2011，968-976. 28600 Mary's Cour t ， E a s to n ， M D 21601US A