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 TCS SP5 MP
LEICA ( 德國 )  的共軛焦多光子顯微影像技術 與 設計, 率先採用光纖傳導雷射與 EOM 雷射調控, 以及, 專利的超靈敏 NDD 光譜偵測器, 領先其他廠商, 迄今, 尚無其他廠商可以趕上媲美.

今日的 TCS SP5 系列, 也是唯一全光譜 ( Excitation / Spectral Beam Splitter / Emission -  Spectral Detection ) 可調式的共軛焦顯微鏡系統, 更是未來 超光學解析   ( Super-Resolution ) 技術與系統的基礎平台. 結合全世界最先進的光學技術, 邁入單分子偵測影像的領域 ( dynamic single molecular detection ), 創造了許多令人激賞與期待的結果.

Leica TCS SP5 MP 多光子影像系統, 是架構在 TCS SP5 II 上, 可升級採用 Ti:Sapphire 飛秒雷射 ( Femto second 或 Pico second ), 因為減少光的吸收及激發光的散射, 所以, 可減少光漂白及光毒害, 可做厚樣品的深層影像擷取., 例如活體的細胞, 器官, 胞器, 組織.. 等樣品.
一般多光子影像的應用領域

Neuroscience

  • Calcium Dynamics in Brain Slices

  • Neuronal plasticity

  • Synaptic physiology

  • Spine structure-function relationship

  • Dendritic signalling

Immunology

  • Lymphocyte trafficking

  • Immune system dynamics

Development

Pathological conditions
(Development, progression and potential treatment)

  • Tumors (angiogenesis / metastasis)

  • Alzheimer disease

Diverse specimens:

  • Lymphatic organs

  • Kidney

  • Heart

  • Skin

  • Brain
TCS SP5 II 光譜式共軛焦顯微鏡系統 是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.
多光子影像技術的好處 ...
  • 降低雷射激發光的散射, 所以更能深入穿透樣品, 適於厚樣品的應用.
  • 激發光僅限於焦平面的極小點, 不是整個大面積的區域, 減低雜訊. 避免快速螢光漂白.
  • 無須使用 Pinhole
  • 減低光毒害, 適合活細胞影像擷取.
  • 極適合應用於 uncaging, photoactivation, Photobleaching

Excitation:
 - No MP excitation outside of  focus

 - Simply increase excitation  intensity to compensate  scattering losses deep inside    sample

Emission:

- No out-of-focus-fluorescence
- No confocal aperture needed      
- Use non-descanned detection to 
  capture all scattered photons

　
Leica 多光子影像系統採用超靈敏光譜訊號偵測器 ( NDD ) , 具有專利的設計. 具有最高 4 通道設計組合. ( 4-channel NDD )

  • Scattered fluorescent photons can also be collected

  • Special dichroic allows simultaneous acquisition of fluorescence and IR-SGC
NDD ( Non-Descan Detector ) RLD (反射螢光偵測器  ) TLD ( 穿透光偵測器 )
2 ch. / 4 ch. NDD

Detectors directly behind Objective, RLD

Detectors directly behind Condensor, TLD
　 　 　
mean intensity image : 20

  • Mouse brain slice:    ~ 20 µm (center plane)

  • Detection range:      500 – 550 nm

  • PMT:             950 V

  • Objective:                 20 x 1.0 W

  • Excitation:     920 nm, power level identical 
Leica TCS SP5 MP 的 Ti:Sapphire 雷射光源 ( 690 - 1040 nm ) - Leica 獨家 EOM 調控
Newport ( Spectra-Physics ) Coherent

High power (HX) category ( 高功率 飛秒雷射 )

  • IR Laser MaiTai HP, ps,690-1020nm

  • IR Laser Chameleon Ultra, ps, 690 - 1020 nm

  • IR Laser Chameleon ultra 1, ps, 690 - 1040 nm

  • IR Laser Chameleon ultra II, ps, 680 - 1080 nm

Standard power category ( 常用一般功率 飛秒雷射 )

  • IR Laser MaiTai, ps, 780 – 920 nm

  • IR Laser MaiTai wideband, ps, 710-920nm

  • IR Laser MaiTai broadband, ps, 710-990nm
Leica 獨家多光子影像應用專屬物鏡
物鏡是光學解析的重要因素, 其影響實驗好壞與解析因素包括物鏡倍率, NA 值, 工作距離, 解像力  ( XY / XZ ), 浸漬媒介物 ( 空氣, 水, 甘油 )

  • Optimized for 3D-imaging of thick specimen

  • HCX PLAN APO 63x 1.3 CS

  • Immersion: 80% glycerol and 20% water

  •  Free working distance of 280 µm

  •  Correction collar for

  • coverslip thickness

  • immersion medium variations

  • temperature

  • Optimized for temperature: 21 °C
　 　

HCX APO 20x 1.0 W       A New Objective for Electrophysiology

  •   Low magnification 20x

  •   High resolution by high NA 1.0

  •   2 mm free working distance

  •   Access angle 39° for good sample accessibility

  •   Ceramic non-conducting lower part for

  •   nterference-free patch clamping

  •   High transmission in VIS-IR:

  • Image dimension @ Zoom 1 : 7752 µm

  • Resolution xy (488nm):         195 nm

  • Pixel Size acc. Nyquist:         97.5nm

  • Pixel size @ Z1 & 81922:      95nm

 

  • Imaging of primordial germ cells (PGC) in Drosophila embryos

  • Visualisation with confocal imaging very difficult

  • Depending on stage egg yolk typically „masks“ the PGCs

  • Egg yolk hardly penetrated with visual light

  • PGCs often only visible with IR illumination

多光子影像  應用 ( 長時間紀錄電影檔, 請點閱圖示 )
　

  • 3 channel overlay with RNA injection H2B (blue),
    m cherry (red)      EGFPras (green)

  • conventional scanner- 200Hz, 512x512 ,

  • MP excitation

  • 420 stacks

  • Time-laps: 21H

 　

  • Time lapse deep in-vivo Imaging,  Excitation at 890nm , 3 channel acquisition:

  •  Autofluorescence of elastin (blue),  Syto13 for nuclei of cells in the vascular wall (green/white),  Eosin auto-fluorescence (red);

  • Imaging depth 650µm.

  • Preparation

  • Common carotid arteries from mice are carefully dissected, excised, and stored in Hanks’ balanced salt solution (HBSS, pH 7.4). 
LEICA TCS SP5 II  圖示 ( 簡潔極短的光路, 運藏許多創新的專利設計, 達成最高的光學解析與使用效益.
LEICA TCS SP5 II ( 架設於倒立顯微鏡 DMI 6000 B ) LEICA TCS SP5 II ( 架設於正立顯微鏡 DM 6000 B )
使用於 TCS SP5 II 的基本雷射系統 ( 選配 )
Continuous wave lasers Laser Type For Imaging
VIS WLL, average power 1.5 mW, 470 - 670 nm
Diode, 40 mW, 442 nm
Ar, 65 mW, 458, 476, 488, 496, 514 nm
HeNe, 1 mW, 543 nm
HeNe, 2 mW, 594 nm
HeNe, 10 mW, 633 nm
DPSS, 20 mW, 561 nm
UV UV OPSL, 80 mW, 355 nm
Pulsed lasers IR TiSa ( femto second or pico second ), 1 W, 690 - 1040 nm ( various range )
共軛焦 與 原子力顯微鏡 的整合 ( Confocal + AFM )
Leica TCS SP5 II equipped with AFM ( Veeco Bioscope II )

AFM image

Wild type of purple bacteria Rhodobacter Sphaeroides

S. Bahatyrova et al. Nature, 430, 1058-1062 (2004

1. High resolution imaging of biomolecules under physiological conditions in real-time and in situ (static and dynamic/time-lapse imaging)
　

  •  Nucleic Acids
  •  Proteins
  •  Membranes
  •  Cells
  •  Tissues
  •  Biomaterials

2. Single molecule measurements of inter- / intra-molecular interactions and nanomechanical properties
　

  •  Protein-Protein / Ligand-Receptor Binding
  •  Molecular (Un)Folding
  •  Surface Elasticity / Adhesion

　

3. Mechanical manipulation of tissues and intracellular elements 

TCS SP5 MP 架構於  TCS SP5 II 共軛焦顯微鏡系統, 可使用於正力或倒立顯微鏡, 僅需雷射光纖管切換. 全方面的設計, 也可升級到超解析 ( Super-Resolution , STED, SMD-system, FLCS, FLIM, FCS ... etc ) 的系統, 滿足未來的需求.
請下載檔案 .. 　
　

Downloads Leica TCS SP5 MP

Application Notes

EPhysSoftware_AppLetter.EN (PDF, 1013.33 K, 04. Mar 2009)
Ephys_DeepTissue-AppLetter.EN (PDF, 2.96 MB, 19. Nov 2008)

Brochures

Leica TCS_SP5_II-Brochure.EN (PDF, 3.11 MB, 07. Jul 2009)
Leica TCS_SP5_II-Brochure_Technical_Data.EN (PDF, 389.92 K, 07. Jul 2009)
Leica_TCS_SP5_MP-Flyer.EN (PDF, 1.75 MB, 27. Nov 2007)
TCS SP5_System Overview.EN_June09 (PDF, 1.29 MB, 21. Dec 2009)
　		 　

Application Notes

Appl_Let_17_Glycerol_Objective_144dpi (PDF, 1.33 MB, )
Dye Finder-AppLetter_30.EN (PDF, 3.99 MB)
EPhysSoftware_AppLetter.EN (PDF, 1013.33 K)
Ephys_DeepTissue-AppLetter.EN (PDF, 2.96 MB,)
FRAP-Application_Letter_21.EN.pdf (ZIP, 1.02 MB)
FRAP2-Application_Letter_23.EN.pdf (ZIP, 224.44 K)
FRET_AB-AppLetter_28.EN (PDF, 1.76 MB)
FRET_SE-Application_Letter_20.EN.pdf (ZIP, 949.05 K)
LiveDataMode-AppLetter_27.EN (PDF, 2.31 MB,)
Photoconversion-Application_Letter_22.EN.pdf (ZIP, 222.93 K,)
SP5 X WLL-AppLetter_29.EN (PDF, 4.59 MB)
STED-AppLetter_32.EN (PDF, 6.29 MB, )
Leica_DM6000 CFS-Flyer.EN (PDF, 692.04 K, )
Leica_TCS_SP5_MP-Flyer.EN (PDF, 1.75 MB, )
SP5 AOBS-flyer.EN (PDF, 770.55 K,)
SP5 Resonant Scanner-flyer.EN (PDF, 1.97 MB, )
SP5 SP Detector-flyer.EN (PDF, 892.51 K, )
SP5 Tandem Scanner-flyer.EN (PDF, 966.94 K, )
TCS SP5_System Overview.EN_June09 (PDF, 1.29 MB, )

Leica TCS SP5 II 目錄 ( PDF, 3042 KB ) 　
Leica TCS SP5 X 目錄 ( PDF, 578 KB ) 　
Leica TCS SP5 II Technical Datasheet ( PDF, 381 KB ) 　

Leica TCS SMD series 目錄 ( PDF, 3768 KB )
　
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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