(Lecture) How to Use Laser Scanning Confocal Microscope SP8 (Lecture (in English))

山元 武文・森 康博(実験実習支援センター)
Takefumi Yamamoto, Yasuhiro Mori (Central Research Laboratory)
Collaborator: Leica Microsystems K.K.

 Leica TCS SP8は倒立型共焦点顕微鏡(全自動蛍光微分干渉)システムで、プレパラートサンプルだけでなく、φ35 mmカバーガラスボトムディッシュ上のLivingサンプルも観察可能です。励起レーザーとして405 nmレーザーおよびWLL (White Light Laser; パルスレーザー)を搭載しています。WLLは470 nmから670 nmまでの任意の波長を最大8波長まで同時に発振することができるため、試薬・蛍光タンパク質に最適な励起波長を選択することができます。
 超高感度検出器HyD (Hybrid Detector)を搭載しているため、これまで画像取得が困難であった微弱なシグナルやダメージに弱い生細胞もS/Nの良い高解像でイメージング可能です。また独自のプリズム分光方式により、試薬・蛍光タンパク質に最適な蛍光波長の選択やクロストークの軽減、自家蛍光の除去なども実現できます。

 Leica TCS SP8 is a fully automated inverted confocal microscope system including DIC, can observe both slide glass (prepared slide) and living cells on the φ35 mm dish (Dish with glass bottom). Equipped lasers are 405 nm and WLL (White Light Laser; pulsed laser). WLL excites any wavelength between 470 nm and 670 nm and up to 8 excitation lines simultaneously. Then, WLL perfectly matches the wavelength of any fluorophore.
 It is also possible to detect dimmer or photo-sensitive specimens since a HyD (Hybrid Detector; super-sensitive detector) is built in. These kinds of specimens were somehow difficult to acquire, however HyD enables you to acquire image in high signal-to-noise ratio and resolution. Easy to avoid cross talk and possible to separate from auto fluorescence by using our unique prism-and-slit detection method for fluorescence.
 Thanks to the prism-and-slit detection and WLL, two-dimensional excitation-emission spectrum is available. Thanks to the WLL and HyD, non-wanted fluorescence can be removed.

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Last Updated 2020/7/27