1、Microscopy from Carl ZeissLSM 780Get More ResultsThe New Detection Quality in Confocal Laser Scanning Microscopy.Get More Results Through Universal DetectionJust as you aim to advance scientific knowledge through your research, we are striving to equip you with the tools to make that breakthrough po
2、ssible. Universal detection yields more results to drive forward your research in neurobiology, physiology, and develop-mental biology. This is ideal for cell and molecular biology too.2Title:Innervation of dorsal body wall muscle next to the heart of Drosophila melanogaster larva. Red: anti-Spectri
3、n staining. Green: GFP expressed in the heart. Ventral view. Sample: J. Sellin, University of Osnabrck, Germany.Right:Embryonic Drosophila melanogaster, Alexa anti-FP staining of brain structures.Weve worked closely with leading scientists worldwide to create an instrument that reflects the latest i
4、deas and technological possibilities. 34The LSM 780 with LSM BiG on upright Axio Imager or Axio Examiner.5The LSM 780 on inverted Axio Observer. 6The GaAsP detectors allow integration and counting detection modes.Sensitivity, Spectral Imaging and Photon CountingChallenging samples require more than
5、just amplifying weak signals. You need a detector thats capable of dealing with high signal dynamics, provides low noise and allows for short pixel times.The more demanding the application in laser scanning microscopy, the greater your need for sensitivity and reduced background noise. Thats why we
6、have taken the proven sensitivity of the LSM 710 an important step forward with the LSM 780s GaAsP spectral detector.The group of 32+2 detectors allows you to reproduce spectral measurements reliably and without deviation. Since this is a parallel spectral detection design, it offers you simultaneou
7、s 34-channel readout in lambda mode. Up to 10 dyes can be acquired and separated at the same time. With the new GaAsP detectors, you get up to 2x better SNR for 2x faster acquisition.With it comes a whole range of performance-enhancing improvements: 32-Ch GaAsP detector with 45% Quantum Efficiency (
8、QE) typically Plus photon counting ability on the GaAsP and new side PMT Great dynamics especially to visualize weak signals Active cooling and oversampling photon counting mode for best SNR“We havent been able to image yeast cells on a confocal yet. With the GaAsP detector we can do this now.”“also
9、, very low expressing fluorescent proteins in cultured cells can be imaged in a new quality class.”“The GaAsP detector allows to speed up acquisition by a factor of two, with the same superb image quality as usually achieved by averaging twice on the PMTs.”“Its a great system, we really like the ima
10、ge quality.”“ROI-HDR is extremely useful, especially it is implemented in such a flexible and intelligent way.”Comments of LSM 780 and LSM BiG beta testers from labs in France, the United Kingdom, and Germany.7Drosophila melanogaster larvae, developing brain and eye neuronal structures labelled with
11、 three FPs in blue, green, and red.Cultured 2h8 cells labelled with extremely low expressing GFP and mCherry; left PMT (almost invisible), right GaAsP. Sample: A. Bruckbauer, Cancer Research, London, UK.Dividing yeast cells, labelled with eGFP and Tomato. Sample: I. Jourdain, Cancer Research, London
12、, UK.8Integrated FCS and FCCSFluorescence Correlation Spectroscopy (FCS) lets you ana-lyze single molecules at a new level, using its revolutionary GaAsP detector. Up to 6 channels can be used in FCS mode, providing great flexibility in your stainings and samples.Until now, this technology required
13、expensive external detectors. With its internal spectral detector, the LSM 780 is able to perform FCS analysis with an actively cooled, photon counting GaAsP detector.Until now, your choice of a high sensitivity detector was limited by its ability to deal with the color of dye you were using. As a r
14、esult of the spectral capability of the GaAsP detector, the dyes you choose can now be located any-where in the whole color spectrum. That makes the LSM 780 much more versatile. Until now, you had to compromise between very strong and very weak signals. The GaAsP detector is highly sensi-tive toward
15、s light and has greater dynamics as e.g. APDs, enabling you to deal with both extremes at the same time without extra settings. As a result, you save time and data volumes. 0 s 10 s 20 s 30 s 40 s0 s10 s20 s30 s40 s0 s10 s20 s30 s40 s040 s0 s10 s20 s30 s40 s0 s10 s20 s30 s40 sGrafikThanks to its exc
16、ellent sensitivity and counting ability, the LSM 780 lets you tap into the power of integrated spectral FCS (Fluorescence Correlation Spectroscopy) to analyze single molecule dynamics. Snapshot of diffusion of particles. Spatial and time correlation can be analyzed to obtain number and speed of the
17、molecule populations.Areas with the best count rate or interesting regions can be chosen in the image.Auto- and cross-correlation analysis using the GaAsP detectors.9Quantitative Imaging ExtensionsThe spectral GaAsP detector of the LSM 780 can also be retrofitted to the LSM 710, bringing this great
18、technology to your lab. But there are more powerful methods that can extend the data from your sample. Even without the internal GaAsP detector, LSM 710 and LSM 780 systems offer possibilities that go beyond conven-tional imaging. The detectors in every LSM 710 and LSM 780 system offer Raster Image
19、Correlation Spectroscopy (RICS), a method developed by E. Gratton and P. Wiseman for measuring fast protein dynamics and concentrations. RICS requires no special hardware detectors: its analysis is done in the normal scanned image and provides precise analysis of many fast-moving molecules.Anisotrop
20、y imaging is another method that offers you an additional parameter of the emission light to investigate proteins: polarization. Because fluorescence polarization and hence anisotropy will vary according to the distance and bond of the molecules, this method tells you about the spatial proximity pro
21、perties of the molecules in your sample (e.g. FRET). The polarization filters required for anisotropy can be supplied with, or retrofitted to any LSM 710 or LSM 780.While the internal GaAsP detector is exclusive to the LSM 780, you can also enjoy many of its advances by adding the LSM BiG (binary Ga
22、AsP) external module to the direct cou-pling port. This brings authentic GaAsP performance to your LSM 710 system with its two channels allowing both sensi-tive imaging and FCS analysis.Actin filaments labelled with Alexa Fluor 488-phalloidin in the Drosophila eye, showing the Anisotropy. Only in so
23、me rhab-domers filaments are similarly oriented. Sample: O. Baumann, University of Potsdam, Institute for Biochemistry and Biology, Germany.RICS image of GFP labelled U2OS cells with display of correlation in 2.5 D (big window) and diffusion map (bottom right). Specimen: U. Schmidt and E. Bertrand,
24、IGMM-CNRS, Montpellier, France.10Nuclei of yeast cells labelled with GFP, fast FRAP/FLIP time series of rapid diffusion. Sample: F. Bollet-Quivogne, Cancer Research, London, UK.Fast OSCiscanThe LSM 780 and LSM 710 are both excellent live cell imaging systems. Now, with the OSCiscan, you have the fas
25、test point scanning solution ever.Fast point scanning is usually restricted to 4-5 frames per second at full formats. Until recently the only way to go faster was in the resonance mode, which allows scanning at one resonance frequency of the scan mirrors. Since this speed is fixed at a non-ideal pix
26、el time, the image outcome will usually be compromised by high noise. The LSM 780 and LSM 710 have now overcome this prob-lem via the OSCimode, which allows 8 frames per second at full format or 250 fps at 512x16 pixels. The key to the OSCimode is the Online Scanner Calibration, where the position a
27、nd movement of the scan mirrors are corrected on the fly during scanning. This achieves a perfect linear movement at extremely high speeds, along with the ability to choose the precise pixel time and speed your sample requires.The absence of any annoying resonance sound is an added bonus.Freely defi
28、nable regions of interest (ROIs) are essential for bleach and photo activation experiments, whether they in-volve cancer research, cell death, the analysis of DNA repair proteins, protein synthesis, or the detailed mechanisms of cell division. Both the LSM 780 and LSM 710 offer ideal tools for manip
29、ulation of single and multiple ROIs with in-dividual settings at the fastest speeds possible.Developing Drosophila melanogaster, eye differentiation labelled with GFP e-Cadherin, 4D time lapse acquisition with fast Z-stacks. Sample: E. Chan, Cancer Research, London, UK.11The number of frames and ill
30、umination variations can be set freely for the HDR acquisition.ROI-HDR ImagingLess damaging laser powers and higher detection dynamics yield more valid results in live cell imaging.High dynamic range (HDR) imaging is well known from still photography. In a live imaging system, it seems to be a bad i
31、dea to obtain multiple images and expose your sample to repeated laser illumination. However, in biological samples the fluorescence dynamics are often bigger than the capac-ity of the detection system. The ROI-HDR mode of the LSM 780 and LSM 710 allows you to image the weak and bright portions of a
32、 frame in the first shot (or line) and then acquire the weak signal subregions (ROIs) with a second shot to amplify them. The software lets you choose between an adapted mode or a linear mode which even allows quantita-tive analysis of the signals.The result is an increased dynamics of the detection
33、, with-out superfluous laser exposure to the sample. Without satu-ration, for example, a bright soma of a nerve cell can be imaged together with the weak and faint dendrites so its altogether easier on your specimen. Non-neuronal retina structures labelled with Cyano-dyes, very high signal dynamics;
34、 left conventional, right HDR acquisition. Sample: F. Tatin, Cancer Research, London, UK.12Ideal Acquisition Strategies with Smart SetupBy choosing the right acquisition strategy, you can use more dyes than ever before without crosstalk. Smart setup allows automatic setting of imaging parameters dep
35、en-ding on your preferences for acquisition speed and signal quality.Frontal section of Drosophila melanogaster brain, triple antibody labelled for synapses and neurons.Carl Zeiss offers a unique tool to improve your imaging: the Smart Setup function. With the spectral properties of several hundred
36、dyes known by the system, Smart Setup can recommend an acquisition strategy that will increase acquisition speed or signal outcome without crosstalk, de-pending on which dyes you use. This knowledge database is constantly updated.One valuable side effect is the excellent training those who are less
37、experienced in imaging facilities will get on the properties of their samples and how to set up a modern confocal system.13In Tune and TwinGateUse the latest dyes with extreme spectral properties and obtain lifetime data at any wavelength.The innovative TwinGate low angle main beamsplitter pro-vides
38、 up to 100 combinations of excitation laser lines which you can exchange at will. The lasers including pulsed lasers and powerful bleach lasers can be combined freely from near UV (355, 405 nm), VIS, and IR (Ti:Sa) ranges. On the detection side, emission bands can be flexibly selected without emissi
39、on filters or secondary dichroics.The LSM 780 is prepared to accept new 355 nm DPSS UV lasers which will be available later in 2010. This allows you to image UV excitable dyes without sacrificing the blue-green detection range. On LSM 710 systems, the optics for such lasers can be retrofitted.Mouse
40、kidney section stained for podocyte (Alexa 488, green), membrane Nidogen (Alexa 555, red) and Nuclei (Topro-3, blue). Sample: B. Hartleben, R. Nitschke; University of Freiburg, Medical Clinic IV and Life Imaging Center, Germany.The fast and flexible detection technology of the LSM 780 and LSM 710, c
41、ombined with our high performing In Tune laser, gives you additional freedom in excitation. This flexibil-ity in excitation (488 to 640 nm 1.5 mW per wavelength) means the fluorescence signal can be detected very close to the excitation wavelength. In Tune can be used simultane-ously with any laser
42、available in the system.This is the perfect flexible laser system for measuring flu-orescence lifetimes of dyes (Pulse 5 ps, 40 MHz) that couldnt be examined before. Also, you no longer need to compromise when searching for a FRET pair. In Tunes wavelength range lets you measure the lifetime of any
43、dye excited within the spectral range of 488 to 640 nm. In Tune wavelengths can be selected easily from a drop down list, while the matching MBS is set automatically.14NLO with LSM BiG NDDMultiphoton imaging puts a powerful technology at your disposal because not only physiologists and neurobiologis
44、ts need to be able to get extended depth imaging of three-dimensional samples.LSM BiG non-descanned detector (NDD) with 2 GaAsP channels on the Axio Observer.Projecting neurons in Drosophila melanogaster, antibody triple staining showing synaptic connec-tivity.The new GaAsP detector technology is no
45、t confined to visual light excitation. The LSM BiG (binary GaAsP) now also of-fers you multicolor multiphoton imaging with GaAsP perfor-mance. Two LSM BiG modules can be added to NLO systems as transmitted and incident light NDDs, providing 4 ultrasensi-tive detection channels. The LSM 780 NLO and L
46、SM 710 NLO let you penetrate deeper and detect more light.Improved femtosecond multiphoton technology lets you move from flat caricatures into a three-dimensional con-text so you can understand interrelations in complex bio-logical systems. The enhanced sensitivity of the BiG helps you penetrate eve
47、n deeper into your samples.Together with our multiphoton special objectives W Plan-Apochromat 20/1,0 and C-Achromat 32x/0,85 W IR, you get ideal solutions for nonlinear optical (NLO) imaging. In addition to the Axio Examiner stand for physiologists, cell biologists can also use such IR objectives an
48、d the LSM BiG on the inverted Axio Observer stand.15NLO and Uncaging with OSCiscanAs a physiologist and neurobiologist, you need powerful imaging to analyze the interaction of cells in a tissue context.Versatile multiphoton imaging with GaAsP detectors.Synapses and selected neurons in Drosophila mel
49、anogaster, labelled with Alexa anti-xFPs. Multiphoton imaging with PMT-NDD detector (left) and internal spectral GaAsP detector with open pinhole (right).The LSM 780 NLO and LSM 710 NLO offer you all the ingredients you need: efficient multiphoton imaging with specialized objectives, short beampaths and LSM BiG plus fast scanning with the OSCimode and additional uncaging or photomanipulation.This package allows you to, for example, uncage neuro-transmitters at the synapses and image the reaction of the cell deeper in the tissue via IR wit
