高階多維度影像分析平台

High-Dimensional Advanced Imaging Core Facility

I6-正立型共軛焦顯微鏡 -共軛焦系統

Leica SP8 Deep In Vivo Explorer (DIVE) /Confocal

Leica SP8 Deep In Vivo Explorer (DIVE) /Confocal

正立型共軛焦顯微鏡 -共軛焦系統
正立型共軛焦顯微鏡 -共軛焦系統
正立型共軛焦顯微鏡 -共軛焦系統

儀器描述

Leica SP8 DIVE為全球第一套全光譜自由多色、多光子深度活體影像系統。本系統配置可調式倍頻雷射,可做共軛焦顯微鏡與組織深層之穿透掃描。經過特殊閃電(Lightning)運算方式,解析度亦可達120 nm。搭配最新型可調光束擴展器 (VBE):可靈活調整光束聚散,在解析力及觀察深度取得絕佳平衡。

主要特點

  • 單光子共軛焦掃描:
    多色高解析度觀察 (Ligtening)
    多點位置應用及組織樣品大面積拼圖
    弱螢光影像
    澄清化厚組織樣品或透明樣品 (如:透明斑馬魚,細胞層級等)
  • 多光子共軛焦掃描:
    活體動物/厚組織樣品,增加取樣的深度,能更深入樣品裡層
    減少光毒害,減低螢光漂白速率,增加樣品螢光觀察與掃描的時間
    無須感測光針孔 (pinhole), 反射螢光可 100 % 被偵測

儀器規格

  • 電動正立型顯微鏡:Leica DM6 CFS flex
  • 雷射系統:
    共軛焦系統:405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 594 nm,及633 nm
    多光子系統:可調式雷射660-1320 nm, 固定波長1040 nm
  • 螢光濾片:DAPI, FITC, TexasRed
  • 物鏡:
    10x, 20x甘油鏡, 40x油鏡, 63x油鏡, 100x油鏡, 及20x Imm Corr,
    25x (IR專用水鏡, NA 1.0, 工作距離: 2.6 mm)
  • 感測器:
    共軛焦系統:兩組共軛焦顯微鏡專用PMT感測器,一組HyD高感度感測器
    多光子系統:兩組專用4Tune NDD高感度感測器
  • DIC 微分干涉相位差觀察模式
  • 電動載物台
  • 控制軟體:LAS-X SP8
  • 離線分析軟體:LAS-X SP8

使用注意事項

  • 建議使用1.5H (0.17 mm) 蓋玻片
  • 使用Multi-photon需先經過共軛焦基礎訓練後再次考核多光子系統操作
  • 以下功能應用不提供自行操作:
    1. 非一般標準玻片型式之樣品,非乾式樣品,具有溶液或細胞培養液樣品。
    2. Photon manipulation實驗

放置地點

國家生技研究園區A棟5樓502b室

管理人員

羅麗紋 博士

電子信箱:llw@gate.sinica.edu.tw

電話:02-7750-5554 或 02-7750-5556

Description

Leica SP8 DIVE (Deep In Vivo Explorer) is the first multiphoton microscope with spectrally tunable detection. The SP8 DIVE provides spectral freedom: Equipped with 4Tune, a tunable, non-descanned detection unit, the SP8 DIVE offers you unlimited flexibility and enables you to develop new multicolor deep in vivo experiments. The SP8 DIVE provides maximum penetration depth and best contrast: The new Vario Beam Expander can be tuned for improved penetration depth of more than 1 mm and maximum resolution. Achieve more contrast and depth for multicolor, deep in vivo imaging. The SP8 DIVE gets the best out of your experiment!

Application

  • Confocal imaging:
    Multi-color high-resolution scanning -LIGHTENING
    Multiple position application and large-area tile scanning
    Low-fluorescence imaging
    Clearing tissue/sample (large scale tissue/ Zibrafish…etc)
  • Multi-photon imaging:
    Animal/thick tissue imaging
    Decrease light toxicity and photo-bleach could be increase observation and scanning time
    No pinhole, reflection light could be increase

Specification

  • Microscope main body:Leica DM6 CFS flex
  • Laser system:
    Confocal system:405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 594 nm, and 633 nm
    Multi-photon system:Tunable 660-1320 nm, fix-wavelength 1040 nm
  • Filter:DAPI, FITC, TexasRed
  • Objective:
    10x, 20x Imm Corr, Glyc oil, 40x oil, 63x oil, 100x oil, and 20x
    25x (IR water, NA 1.0, W.D. 2.6 mm)
  • Detector:
    Confocal system:2, PMT detector;1, HyD detector
    Multi-photon system:2, 4Tune NDD detector
  • DIC module
  • Automatic sample stage
  • Control software:LAS-X SP8
  • OFF-Line software:LAS-X SP8

Precautions for user

  • Recommend to use “1.5H (0.17 mm)” coverslip
  • To certify of Multi-photon training must be complete the basic confocal operation training.
  • Some functional application is only technician operation:
    1. Non-common used slice, Wet-type sample which contain culture medium and buffer
    2. Photon manipulation

Facility location

National Biotechnology Research Park Building A, 5th Floor, room 502b

Manager

Li-Wen Lo, PhD

E-mail:llw@gate.sinica.edu.tw

Tel:02-7750-5554 或 02-7750-5556