徠卡顯微鏡系統
雷射掃描共軛焦顯微鏡
半導體及工業用顯微鏡
顯微數位影像系統
細胞基因影像分析系統
其它光電微機電系統
其它生醫研究儀器
維修與技術支援服務
聯絡與留言
　
　
　 了解 Leica 在 Super-Resolution 的解決方案 與 領導趨勢 ..
　 　
　
　
多一點了解 Leica TCS SP5 的卓越, 與其他產品的差異性在哪裡 ?
　 　
　
Leica 創造出一個拉曼顯微鏡術的新名詞 , CARS Microscopy =  Vibrational Microscopy
　
Leica TCS CARS 是結合 TCS SP5 II 及 CARS 為一個高速高解析的非線性光學成像平台. 提供了無需螢光標定的分子成像系統.也是未來多模式超解析光學平台的基礎,  例如 STED, STED CW, CARS, SRS, SMD-FCS, FLIM, FLCS ... 等.
　

當我們使用雷射掃描共軛焦顯微鏡 (多光子螢光激發影像擷取), 受測樣品經常要標定螢光染劑 (fluorescence dye ). 然而, 使用螢光染劑, 經常會面臨一些使用上的限制, 包括 :

  • 螢光染劑的分子大於受觀測樣品, 致使訊號嚴重干擾, 會有誤判現象.

  • 螢光染劑容易快速漂白

  • 螢光染劑具有毒性, 會破壞活細胞

  • 螢光染劑的涂染取像過程, 經常會有訊號淹蓋真實的狀況.

  • 使用螢光染劑標定過程耗時

  • 有些樣品不能使用與不容易使用螢光染劑來標定

  • 已經過染劑標定的樣品也不適合長時間觀察, 因為環境會破壞染劑的生理特性,

　
　
Leica TCS CARS ( Coherent anti-Stokes Raman Scattering ) 同調反史托克拉曼散射光譜影像系統, 就是解決此一使用螢光染劑限制的技術之一. 此種技術有下列優點 :
　 　
Leica TCS CARS 是螢光顯微鏡數的拉曼光譜影像

Leica TCS CARS 是隸屬非線性光學影像技術, 兼具高速取像與高解晰取像, 擁有更高靈敏度、更高光學解析度、更低的破壞性、而且具有化學專一性且非破壞性的三維影像, 可以執行深層樣品的觀察, 也可以做斷層掃描.
使用上, 樣品無須標定螢光染劑, 且具有化學鑑別力. 非常有利於脂質的探測，因為碳氫鍵 ( C-H ) 的大量聚集能夠產生很強的特徵信號。( 因為樣品內的化學鍵能夠改變通過其中的光的波長 )
適用於活細胞, 活體組織與器官, 例如在活體組織中對神經髓鞘的監測 ( 因為神經髓鞘緊密的堆積了大量脂質 ), 或者, 探索血管硬化的過程.
適用於長時間影像擷取觀察.
適用各種新的實驗樣品
適合超高解析觀察, 例如可以從細胞的DNA裡分辨出RNA, 利用CARS在細胞中迅速地尋找脂肪體, 討所脂肪酸是否可以用於標記前列腺癌的可能性.
　 　
　
　 LifeCARS image, resolving two types of lipid. Image acquired by Michael Muller, University of Amsterdam.
　
CARS 的技術
拉曼光譜 ( Ramen )
When monochromatic light is scattered by molecules, a small fraction of the scattered light is observed to have different frequency from that of the irradiating light- Raman Effect
Discover in 1928 by Indian Scientist  Sir C. V. Raman, and won Nobel Prize in Physics in 1930.
Ramen spectroscopy is commonly used in chemistry.
　

Different materials has unique Ramen Scattering spectra, a finger print of different  materials.

CARS Microscopy provides chemical contrast generated by Anti-stokes at 4-5 orders magnitude higher than normal Raman Spectroscopy. No FL label required.

The Anti-stokes signal only generated at the focal point-Confocal, allowing 3D sectioning, similar to MP.

The Anti-stokes signal contrast is detected by NDD detectors.
　

　
Optical Parametric Oscillator  ( OPO )

converts input laser (Pump) into two output waves of two waves of lower frequency by means of nonlinear optical interaction.

　
CARS 的應用範例  ( 活體線蟲 )
Life350 x 350 μm2 1,1 s/frame (left), 150 x 150 μm2 0,5 s/frame (right), resonance at 2845 cm-1

全世界唯一全光譜可調式的共軛焦顯微鏡系統, 結合 超高光學解析 與 超高速掃描 於一體. 結合最先進創新的光學解析提升技術, 創建出  Super-Resolution 的基礎平台.
　
其他產品詳細資料, 請點選下列圖示.
　 　
Leica TCS STED Leica TCS STED CW Leica TCS  CARS

Integrates Super Resolution STED with the Broadband Confocal Platform TCS SP5
Leica TCS STED.

With the award-winning invention of STED technology, a new chapter in fluorescence microscopy began 		　 The Fast Track to Superresolution !!! The new Leica TCS STED CW is the most uncomplex way to nanoscopy for research. The system resolves structures smaller than 80 nm – with purely optical methods............ 　 Coherent anti-Stokes Raman scattering (CARS) microscopy overcomes a powerful tool for three-dimensional, label-free imaging of chemical and biological samples in vitro and in vivo.
　 　
Leica TCS SP5 II Leica TCS SP5 X Leica TCS SP5 MP
The Only Broadband Confocal
Leica TCS SP5 II, The Leica TCS SP5 II Confocal covers a broad range of requirements in confocal and multiphoton imaging - with the full array of scan speeds at highest resolution ..........		 　 Tune into any Excitation with Supercontinuum Confocal Leica TCS SP5 X. With the Leica TCS SP5 X Supercontinuum Confocal researchers can tune excitation wavelengths to perfectly match fluorophores, turn down laser power, and thereby increase viability for longer live cell experiments ...... 　 Deep Imaging at Video Rate
Leica TCS SP5 MP...........
　 　
Leica TCS SMD FCS Leica TCS SMD FLIM Leica TCS SMD FLCS
The system acquires and analyzes FCS and FCCS (Fluorescence Cross-Correlation Spectroscopy) data. Both methods focus on quantitative analysis of transport and binding processes, in vitro as well as in vivo 　 It is a dedicated system for FLIM measurements. These measurements reveal information about the direct neighborhood of molecules on the nanometer scale 　 Fluorescence Lifetime Correlation Spectroscopy. This single molecule detection system ensures the highest flexibility in biophysics by conveniently combining FCS (Fluorescence Correlation Spectroscopy) and FLIM (Fluorescence Lifetime Imaging) in one system................
　 　
Leica HCS A Leica TCS SPE II Leica TCS LSI
High Content Screening Automation. The combination of high-resolution true confocal point scanning technology with the ingenious LAS AF MATRIX M3 automation software for the first time offers high content screening on research microscopes. 　 The Leica TCS SPE confocal is a true point-scanning, spectral system at an affordable price for fluorescence imaging of live or fixed cells ........... 　 Capture Intra-vital Adult and Embryo Images from Macro to Micro with Leica TCS LSI. The first super zoom confocal that offers high resolution plus a large 16mm field of view for in vivo imaging
　 　
VT infinite 3 Pinhole array Confocal Scanner 　 Leica Structured Illumination Microscope 　 Leica 4Pi

Next-generation multi-point confocal scanner. Offers low photo-bleaching with high-speed image acquisition .......

 　

Structured Illumination solution allows image contrast improvement, image blur removal and optical sectioning in fluorescence applications

　 A innovative solution for Super-Resolution .......
   相關資料 ( Information )
雷射掃描共軛焦顯微鏡技術概論與應用介紹 ( CLSM Technology & Application )
Leica TCS 光譜式共軛焦顯微鏡的核心技術介紹 ( AOTF, AOBS, Spectral Detector )
Leica TCS 光譜式共軛焦顯微鏡的雷射光源 ( Laser system ) 介紹
Leica TCS 光譜式共軛焦顯微鏡的物鏡 ( Objectives ) 介紹
Leica TCS 光譜式共軛焦顯微鏡的應用軟體 ( Application Software ) 介紹
使用於共軛焦顯微鏡上的活細胞培養設備 ( Micro-incubation )
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