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Leica
TCS SP2 RS
High
speed Confocal & Multi-Photon Live-Cell Imaging
& Recording System 高速雷射掃描共軛焦光譜顯微鏡系統
及 多光子雷射影像系統 (
活細胞影像紀錄系統 ) |
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TCS
SP2
RS
高速雷射掃描共軛焦分光光譜顯微鏡.
建立了共軛焦科技的另一里程碑.
超高速掃描 (
Point-scan ), 適用活細胞高速影像擷取與光譜訊號紀錄
Point-Scan 提供超高解析
( 比
Nipkow-disk 方式更能得到較高的解析
)
可以同時 (
Real-Time ) 做 Multi-channel
螢光影像擷取
光譜式共軛焦系統,
可同時安裝 405
nm 雷射 及 多光子雷射 (
Multi-Photon )
即時穿透光影像 (
Phase contrast, DIC ) 高速掃描
( 多重參數掃描組合
)
激發光設計 ( Excitation )
( TCS SP2,
SP2-AOBS, SP2 RS )
完全採用可程式光波調控設計
( AOTF, Acousto Optical Tunable Filter, programmable
), 包括紫外光雷射,
可見光雷射,
紅外光雷射,
皆採用 AOTF
調控雷射光波選取與強度控制.
釋放接收光譜設計
( Emission ) (
TCS SP2, SP2-AOBS, SP2 RS )
完全採用可程式光波調控設計
(
SP, Spectrophotometer, programmable )
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Leica TCS SP2 RS light path |
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詳細
Leica TCS SP2 RS 請點選連結. Leica
SP2 RS Brochures ( 0.19 MB PDF ) |
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Leica SP2 RS ( Confocal
Spectral Microscope ) 技術原理 |
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| Leica
TCS SP2 RS ( 安裝於 Leica
DM LFSA Fixed stage microscope ) |
Leica
TCS SP2 RS 架設於 Leica
DM IRBE 倒立顯微鏡.
( 活細胞螢光離子流應用,
多光子雷射系統採用
Tsunami
Ti:Sapphire Femto-second 雷射,
Direct-Mirror Coupling )
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Leica
TCS SP2 RS 在光學系統設計上,
與 TCS
SP2 具有相同的設計.
然而,
TCS SP2 RS 具有不一樣的掃描鏡模組,
超高速掃描的 Resonant
Scanning RS-scanner
可達每秒掃描近
8000 條掃描線,
等同 150
fps ( frame per second ) - 512 x 32 Frame size,
X & Y-bi-direction scan mode )
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TCS SP2 RS 掃描速率 |
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同步 Trigger
IN & Trigger OUT
(
Multi-Channel )
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| 溫控及二氧化碳培養系統,
專工長時間活細胞培養與影像擷取紀錄用.
( 可同時銜接 IR-Patch
clamp, Patch-amplifier 或
Micromanipulator
)
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多光子雷射 (
SP Maitai, Femto-second, Direct-Mirror Coupling
)
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光柵
( Slit ) :
位於每一個
PMT 之前,
取代螢光濾鏡與分光鏡.
可任意移位,
可任意移動柵門寬度,
即,
自行設定取決光譜範圍
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| 使用者可自行定義光譜偵測範圍
- 無限制的設定與存取.
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| 使用者可自行定義光譜偵測範圍
- 無限制的設定與存取.
可有效分離重疊的螢光訊號.
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| 無須軟體
UN-Mixing
處理,
即可有效分離幾近重疊的螢光訊號.
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| 可做光譜掃描,
量化螢光實際釋放光譜. |
| Leica
LCS 軟體 |
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唯一的微電腦自動控制調整鏡頭瞳孔照明
( Pupil Illumination )
照明光源能完全穩合所使用的各種鏡頭的瞳孔照明,
是一項極重要的光學解析技術.
簡單的講,
每個鏡頭都有其不同的照明瞳孔孔徑,
一般都是取
40x 鏡頭的瞳孔孔徑當作參考值
( 或其它一個鏡頭的孔徑
), 因而,
忽略了其它鏡頭的瞳孔照明.
如果,
當您使用較低強度的雷射時,
此種沒有穩合瞳孔照明的缺點,
會誤導使用人員,
而深深影饗實驗的工作,
例如,
過度加強雷射強度造成螢光漂白或樣品壞死.,
或者,
錯誤調整感測器靈敏度,
造成影像雜訊過高.
所以,
在可見光及紅外光雷射光源方面,
Leica TCS SP2 已有最佳瞳孔照明設計,
能自動穩合所使用的各種倍率的鏡頭.
在紫外光雷射光源方面,
Leica TCS SP2 已有自動控制的針孔聚光鏡
( Pinhole Lens ).
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| 雷射光源無完全充滿鏡頭的照明瞳孔, 損失光學解析
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雷射光源超出鏡頭的照明瞳孔太多, 損失螢光強度 |
最佳瞳孔照明提供 :
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最佳鏡頭解析 ( 高解像力 )
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最佳照明效益 ( 影像鮮明 )
l
最適宜多光子影像的螢光擷取,
最適宜低功率或低強度的雷射
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此
7 個可旋轉的旋鈕,
每個旋鈕的操控參數,
都可任意選用與設定,
如每一個
PMT 的
Gain 及
offset, Pinhole 的口徑調整,
Z-縱深調焦,
Zoom 變焦,
…..等.
所有軟體的操控,
可設定於此
7 組旋鈕,
可方便使用人員快速的擷取影像與調整最佳的條件.
無須去開啟一大堆視窗,
用滑鼠去推拉一些功能.
往往影像擷取必須快速取得最佳條件,
否則,
過長的時間會造成樣品過度曝晒於雷射下,
樣品壞死,
螢光染劑過速漂白 |
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共軛焦顯微鏡建議使用專屬的鏡頭,
其於
R, G, B, 各作有特別的修正
( correction ), 能提供最高的光學穿透與解析
( Point-spread resolution ). 故, 其有別於一般鏡頭.
例如
: PL-APO 63x/1.32 CS Oil ( 有
CS 標示,
即代表
Confocal System 專屬鏡頭
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| 新一代
405 nm
藍光雷射 (
Blue laser, solid-state Diode blue laser )
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Leica
的 594
nm Laser, 對 Texas
Red 激發效率最佳. |
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Leica TCS SP2 RS
相關技術資料與規格
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Microscope,
Upright, HC |
Leica DM R, RE,
RXE, RXA 2 |
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Leica DM 6000 (
New - 2004 ) |
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Leica DM LFSA (
Fixed Stage Microscope ) |
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Microscope,
Inverted, HC |
Leica DM IRB,
DM IRBE, DM IRE 2. |
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Z drive |
High precision
Galvo-Z-drive : 170 micrometer travel, XYZ : 40 nm
resolution, XZ< 1 nm resolution. |
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Internal
motorization of Leica RE, RXE, RXA2, IRBE, IRE 2
and DM 6000 ( Z-wdie ) |
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Lasers &
Attenuation switching modules |
Ar UV
351, 364 nm |
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He-Cd 442
nm |
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Ar 457 nm, 476
nm, 488 nm, 514 nm ( TCS SP2
RS : new SIRIUS 488 nm laser ) |
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Ar-Kr 488 nm,
568 nm, 647 nm |
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Kr 568 nm |
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CW Solid-State
561 nm ( NEW ) |
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He-Ne 543 nm |
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He-Ne 594 nm (
NEW orange ) |
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He-Ne 633 nm |
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DPSS 430 nm |
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Diode Laser 405
nm ( 20 mW ) - TCS SP2 RS |
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Ti:Sapphire
Lasers ( for Multi-Photon Imaging ) - TCS SP2 RS |
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AOTF, 4 or 8
channels ( for Visible range ) |
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AOTF, UV range |
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EOM, IR range (
Electro-optical modulator ) |
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Laser Merge
Module for 4 lasers. |
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Confocal Unit |
Optics : |
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User-switchable
between Leica microscope ( Upright and Inverted ) |
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Alignment-Free
for lifetime |
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3 individual
ports for external lasers for up to 6 lasers
connected simultaneously |
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Programmable
8-channel AOBS Beam Splitter (
for SP2 AOBS only ) |
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Spectral range
of Detector optics : 400 - 850 nm, programmable. |
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UV and MP
possible in one system ( UV-VIS-MP ) |
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Fiber coupling
( MP : either fiber coupling or direct mirror
coupling ) |
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Field-upgradable
to MP microscopy |
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UV system with
individual objective correction lens |
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Adjustable
PUPIL illumination |
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Single pinhole,
variable diameter size. |
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RS
Scanner : |
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RS
K-scanner with
two independent RS galvanometers. |
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Line frequency
: up to 8000 lines/sec. |
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TCS
SP2 RS Frame rate : 9.4 fps ( 512 x 512 pixels / X
& Y-bi-direction scan ), 150 fps ( 512 x 32 pixels
/ X & Y-bi-direction scan ) |
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Scan resolution
: up to 4096 x 4096 pixels |
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Scan Zoom 1 -
32x ( On-line scan Zoom and
Rotation ) |
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Scan Rotation :
-2 to +92 degrees |
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Scan field : 22
mm diagonal in intermediate image plane |
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Control Unit |
Detectors |
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Highly
sensitivity prism spectrophotometer detector, 1 -
4 simultaneous channels. Up to 8 detector channel
( 4 confocal detector and 4 NDD detector for
Multiphoton imaging ) |
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Continuously
adjustable bandwidth and center wavelength |
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Spectral
steepness factor less than 1 |
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Low noise
photomultipliers |
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Up to 12 bits
digitization per channel |
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Transmitted
light / DIC Detector |
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Non-Descanned (
NDD ) Dual channel transmission detector ( MP ) |
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Non-Descanned (
NDD ) Dual channel Reflection & Fluorescence
detector ( MP ) |
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FLIM detector (
for MP-FLIM or D-FLIM pulsed diode 405 nm ) |
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FCS detector (
for Fluorescence Correlation Spectroscopy ) |
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Optional
External Detector |
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Programmable
Panel-Box, user-configurable |
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Electronics |
Scanner control
system with FRGA ( Field-programmable gate arrays
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Trigger-IN and
Trigger-OUT ports |
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8-Detector
channels, 12-bits digitization, Simultaneous. |
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Ultra-Wide SCSI
interface to PC |
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High
performance PC Workstation ( Windows XP
environment ) |
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One or Two
monitor configuration |
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Software |
Leica LCS
software package |
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Fully
operator-configurable user interface |
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Intuitive and
guided |
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Context-sensitive online help system |
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Multi-dimensional series acquisition ( up to 7
parameters simultaneously ) |
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Programmable
manual control panel |
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Region of
Interest scan |
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Excitation line
/ Frame multiplexing |
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Emission
spectrum recording |
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Multi-color
restoration, spectral un-mixing |
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Advanced Time
lapse recording |
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Surface
reconstruction |
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Multiple
measurement functions |
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Physiology
application software package |
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Multi-Color
application software package |
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FRET
application software |
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FLIM
application software |
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FCS application
software |
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FRAP
application software |
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3D
reconstruction with multiple reconstruction &
rendering functions |
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Macro developer
software |
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De-convolution
software |
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Material
software |
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The 2nd
workstation package ( off-line ) |
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Software new
function |
Leica LCS
MicroLab ( New ) |
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Dye separation |
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Pixel intensity
readout in documentation-viewer |
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Offline phase
correction |
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Zoom-IN
function combined with corp function ( AOTF
control, ROI-scan ) |
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Panning
function during scanning available with Panel Box |
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Objectives |
Leica PL-Fluotar,
PL-APO objectives ( CS ) |
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Live-Cell
Chamber |
Temperature &
CO2 controlled chamber system |
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Wide-Field
Imaging |
Cooled CCD
Camera with software, workstation. |
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Motorized
microscope stage |
Programmable
motorized XY-scanning stage |
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