NIKON ECLIPSE 80i 共聚焦荧光显微镜中文说明书
Nikon 80i荧光显微镜使用说明
Nikon 80i荧光显微镜使用说明△荧光显微镜属贵重仪器，未经保管人员同意请勿擅自使用。
△严格按照操作步骤进行，在使用过程中如发现问题应及时向保管人员反映。
△因违反操作规程，而导致仪器损坏，根据本中心《损坏仪器赔偿制度》进行赔偿。
△使用者在使用完毕后应进行登记.维护和保养本仪器应定期进行维护和保养。
●防止镜头霉变，定期除湿。
●为防止损坏荧光灯泡，荧光光源关闭后，15分钟内不得再重新开启。
●荧光灯泡的最佳使用期限是200小时，如发现荧光强度明显降低应更换荧光灯泡。
荧光显微镜操作步骤△显微镜系精密仪器，务请不要碰击，要仔细耐心使用。
△荧光光源关闭后，15分钟内不得再开启。
△荧光灯泡的最佳使用期限是200小时，因此每次使用时请尽量缩短时间。
1.插上电源插座，打开主机电源开关。
2. 将待检标本置于载物台上。
3. 用10×物镜对焦点，根据使用者的眼间距调节双目镜筒的间距。
4. 调节光圈及灯光强度至合适位置。
5. 调节粗调和微调使标本至最清晰。
6. 拔出活动杆，使光路通过CCD至显示器。
7.保存图象至电脑中。
8. 使用完毕后关掉电源，如镜头上有指纹或污迹用擦镜纸将其擦除。
●荧光光源操作1.插上荧光光源电源插座，打开荧光光源电源开关。
2. 打开光栅。
3. 根据使用荧光素的不同选择不同的荧光滤光片。
4. 将待检标本置于载物台上。
5. 用10×物镜对焦点，根据使用者的眼间距调节双目镜筒的间距。
6. 调节粗调和微调使标本至最清晰。
7. 拔出活动杆，使光路通过CCD至显示器。
8. 保存图象至电脑中。
9. 使用完毕后关掉电源，如镜头上有指纹或污迹用擦镜纸将其擦除。
10. 登记使用情况。
Nikon 80i主要技术参数：放大倍数40-1000，CFI60无限远光学系统，内置滤色镜，数字成像头。
明场、暗场、简单偏光、荧光/明场、荧光/暗场。
波长：330-380，450-490，510-560。
主电压90~250V。
尼康显微镜操作说明
尼康显微镜操作说明尼康A1R共聚焦操作说明第⼀章 A1R共聚焦使⽤总则第⼀条共聚焦操作⼈在⾃主上机前需提交培训申请，经过导师培训，通过考核后⽅可进⾏⾃主上机操作。
第⼆条使⽤共聚焦需⾄少提前1天向仪器负责⼈提交申请使⽤仪器时间段并严格按照规范使⽤仪器。
第三条严格按照规范进⾏开关机，在拍摄过程中如出现任何异常请及时联系仪器负责⼈并说明情况，特别注意仪器使⽤完毕按规范关闭仪器所有开关，清理好镜头，做好登记，保持房间和台⾯整洁，出门时关好房灯和房门。
第⼆章 A1R共聚焦开关机操作说明共聚焦主要包括：显微镜部分、共聚焦部分和计算机部分，请按照以下顺序进⾏开关机。
第⼀条 A1R-si 开机顺序1. 打开 UPS 或接线板总电源开关（⼀般都处于⼀直开机状态）；2. 打开需要使⽤的激光器，不使⽤的激光器可以不打开（具体见后⾯）；3. 依次打开显微镜的长寿命汞灯电源，卤素灯电源，电动载物台控制电源；4. 打开显微镜底座右后端的 HUB 控制部件电源；5. 打开共聚焦控制器左侧⾯板右上⾓的控制器开关；6. 待确认控制器打开的状态下，打开电脑，双击桌⾯ NIS-Elements快捷图标，选择confocal 选项进⼊软件，开始共聚焦操作。
第⼆条 A1R-si 关机顺序1. 请先确认关闭软件，再关闭共聚焦控制器（此条请特别注意）；2. 依次关闭显微镜各个控制部件电源；3. 依次关闭各个激光器之后，关闭激光器电源总开关；4. 关掉 UPS 或接线板总开关（遇到如停电等特殊情况时进⾏此步骤）。
第三条激光器的开关机顺序各个激光器的开关是独⽴的。
下⾯分别加以介绍：⾸先要打开激光器电源盒前⾯钥匙旋钮，然后根据需要打开相应的激光器。
（1）405nm 激光器开机：电源开关拨到 ON 位置-- 钥匙开关拨到 I 位置;关机：钥匙开关拨到 O 位置-- 电源开关拨到 OFF 位置（2）488nm 多线 Ar离⼦激光器开机：确认开关 “标记1”处于 Stand by 位置，钥匙开关“标记2”处于 on 位置，这时风扇开始转动，打开开关“3”，液晶⾯板会有数字显⽰ 17 左右-- 将开关“标记4”由 standby 拨到 operate，液晶⾯板显⽰数值 40 左右，打开完成；关机：将开关“标记4”由 run 拨到 stand by，液晶⾯板会有数字显⽰ 17 左右时关掉开关“3”即可，488钥匙⼀直处于开机状态。
Osa在果蝇卵巢生殖干细胞分化中的功能研究
Osa在果蝇卵巢生殖干细胞分化中的功能研究李敏;瞿杰;李梦婕;胡晓龙【摘要】The Drosophila ovary germline stem cell(GSC)is recognized as an ideal model system for studying stem cell fate regulation in vivo. The epigenetic mechanism is involved in the Drosophila ovary GSC fate regulation. Identification and characterization of apparent epigenetic regulators involved in this process will contribute to reveal underlying regulatory mechanisms. The aim of this study is to investigate the possible functions and the molecular mechanisms of Osa,the chromatin remodeling complex BAP-specific subunit,in Drosophila GSC differentiation. GAL4/UAS binary system combined with RNAi technology was used to specifically down-regulate osa expression in escort cells (ECs),and immune fluorescent dyeing was for detecting the relevant indexes. The results showed that knock-down of osa led to a significant increase of undifferentiated germ cells(UGCs)in the germarium of ovarian tissue,the positive cells of pMad and Dad-lacZ,two BMP signaling activity reporters,increased significantly. Moreover,EC morphology was abnormal,and they failed to wrap germline cells. In conclusion,Osa might be involved in the regulation of GSC differentiation by BMP signaling pathway-depending and specific morphological process of ECs.%果蝇卵巢生殖干细胞(Germline stem cell,GSC)是在活体(in vivo)研究干细胞命运调控的理想平台.表观遗传机制在果蝇卵巢GSC 命运调控中发挥重要作用,其机理的探明需要研究并发现更多参与此过程的表观调控因子.为探究果蝇染色质重塑复合物BAP中特有的亚基Osa在果蝇卵巢GSC分化调控中的功能及其分子机理,利用GAL4/UAS二元表达系统结合RNAi技术在干细胞微环境组分护卫细胞(Escort cells,ECs)中特异性下调osa的表达,并通过免疫荧光染色法对相关指标进行检测.结果显示,敲减ECs中osa可致卵巢组织卵原区中未分化生殖细胞数目(Undifferentiated germ cells,UGCs)显著增多,同时BMP信号通路激活标志pMad、Dad-lacZ阳性细胞数目显著增多,并观察到EC细胞形态异常,不能有效包裹生殖系细胞.推论Osa以BMP信号通路依赖方式参与果蝇卵巢GSC的分化调控,其作用机理还可能涉及EC细胞特定的形态学过程.【期刊名称】《生物技术通报》【年(卷),期】2017(033)009【总页数】6页(P139-144)【关键词】果蝇;Osa;生殖干细胞;分化【作者】李敏;瞿杰;李梦婕;胡晓龙【作者单位】上海交通大学生命科学技术学院,上海 200240;上海交通大学生命科学技术学院,上海 200240;上海交通大学生命科学技术学院,上海 200240;上海交通大学生命科学技术学院,上海 200240【正文语种】中文成体干细胞是一类位于成体组织中的未分化细胞，具有自我更新（Self-renewal）能力与分化潜能，这两方面的特性对于组织内稳态的维持及组织损伤后修复具有重要意义［1］。
激光共聚焦使用手册讲解
10
11. 保存该幅图像: 右击图像管理器中显示的图像图标， 选择另存为保存该幅图像.
(保存为“xml”类型是 FV10-ASW软件专用的 图像格式.)
11
Memo FV10-ASW 专用的图形格式 OIF格式: 创建“一个含 (16-bit TIFF)的图像” 和 “一个附 属文件,”不能单独分割. OIB格式: 创建单个的OIF格式的文件, 方便进行移动和进 行其它的操作.
Note 2 Notes 1 & 3
3. 使用手控面板选择荧光滤色片. (参照 Memo.)
Note 3. 观察之前，检查汞灯的机械快门要放在 开启的位置 ().
3
4. 标本聚焦.
Memo
关于荧光滤色片 NIBA：蓝色激发/绿色荧光 （例：FITC、EGFP等） WIG ：绿色激发/红色荧光 （例：Rhodamine、DsRed等）
3
2
5.
双击快捷方式：
打开 FV10-ASW应用软件. User ID: Administrator Password: Administrator
* 系统软件的启动需要等待一定时间.
5
3
3
显微镜镜下观察
微分干涉差观察
1
手控面板
*DIC 元件
1. 使用手控面板选择物镜.
(参照 Memo.)
10 11
11. 保存该幅图像: 右击图像管理器中显示的图像图标， 选择另存为保存该幅图像.
(保存为“xml”类型是 FV10-ASW软件专用的 图像格式.)
*取消当前荧光染料，选择另外荧光染料， 要双击已指定的荧光染料，并重复步骤2.
点击Apply按钮. (关闭染料选择面板可以用Close按钮.)
尼康荧光显微镜
尼康荧光显微镜技术特点一、光学系统：尼康CFI60无限远光学以其卓越的成像质量受到全球市场的高度好评，齐焦距离60mm。
对人眼观察和数字化成像都是最优的物镜：1、提升的边缘性性和平场度2、消渐光晕（亮度的差别小于5%）3、提升的紫外DAPI 和其他滤光片的焦平面的性能。
二、80i主机的标准配置配备了新的复眼透镜系统，“复眼”透镜阵列使视场边缘的光强达到视场中心光强的90%，所以完全能适用数字成像的照明。
使目镜的视场数达到25mm以上。
三、噪声消除器（H i S/N）是80i 荧光装置的标准特征，信噪比提高了5倍，电动快速低噪声马达滤光片转盘。
四、电动控制装置精巧方便，130W高效长寿命荧光装置，无需调中，滤光片电动快速转换，低噪声，使用寿命2000小时以上，同为NIKON产品，提高产品整体性，方便售后服务五、扩展了人机学性能。
新的矩形机械台：固定位手轮、扭力矩手轮调节，可延伸的工作台固定把手位、超硬铝合金工作台表面。
人机学主机。
显微镜构成尼康显微镜80i的主要构成：（如下图所示）1、80i显微镜主机2、聚光镜3、载物台及夹片器4、荧光物镜：10倍40倍100倍5、物镜转盘6、电动荧光装置：包含荧光专用激发块7、C-BOX2控制器8、双目观察镜筒及照像端口9、目镜10、视频接口11、冷CCD12、长寿命荧光光源主要可调节的部分：（如下图所示）长寿命荧光光纤照明器显微观察方法明场显微术：1 打开电源装置的开关。
将显微镜主机后面的电源开关置于“I”的位置。
电源指示灯亮。
2 按预置开关（显微镜前面的绿色按钮）打开明场照明。
3 将显微镜右下部的ND8、ND32 和NCB11 减光片移至光路中。
（NCB11是白光平衡片，ND8、ND32两个减光片分别将光强减弱为原来的1/8和1/32）4 推入目镜筒上面的光路转换杆，对准双目镜筒使光路充满5 转动激发方式转换盘放在没有激发块的空位处（打开显微镜右后面C-BOX2的开关，通过控制器选择没有荧光块的空位转入光路中）6 提升聚光镜到最高位置。
Nikon 80i显微镜操作流程(荧光)
7、荧光在激发光照射下时间不宜过长，否则易淬灭。
8、未照完的荧光切片可在4度避光条件下短暂保存，不超过24h。
9、数据如何拷贝?是否需要预约?
主要仪器信息
设备名称
正置荧光显微镜
厂家型号
Nikon 80i存放Leabharlann 点A103暗室负责人
联系电话
主要功能
病理切片（HE染色、免疫组化及免疫荧光）的图像采集
使用流程
（荧光）
1、开机：从左往右，荧光显微镜稳压器→图像采集系统→显微镜→电脑。双击桌面的拍摄软件，将其打开。
2、按需要转动荧光挡板。1：蓝色，2：绿色，3：红色。拍照时将SHUTTEER调节至O位置。
7、关闭软件。
8、关机：从右往左，电脑→显微镜→图像采集系统→荧光显微镜稳压器。
注意事项
（荧光）
1、荧光显微镜稳压器开启后15min内不得关闭，关闭后3min内不得开启。
2、不要转动粗准焦螺旋内侧的粗调焦扭力矩调节手轮。
3、采集荧光图像时，不得调节白平衡。
4、图像如需后期分析，曝光时间需一致。
5、修改默认目录：File→options。
3、把载玻片放在载物台上，调节聚焦。在镜下用肉眼观察确定拍摄位置，拉出活动杆将视野切换到电脑。
4、在控制窗口点Exposure，观察预览图像的亮度。选择适当的曝光时间预览图片的亮度合适。
5、点Capture按钮拍摄，等待几秒后保存图片，格式为.JPEG或tiff。
6、点Live按钮，继续拍摄下一张照片。
Nikon 80i正置三光源显微镜操作规程
一、透射明视场观察 开机： 1、打开显微镜电源，按下镜座前端电源按钮，将卤素灯调节到合适亮度。 2、确认光路：目镜筒右侧拉杆全部推入，聚光器转盘在“A”档，荧光滤色块转盘在空档， NCB 滤光片被置入光路，其余 ND 置出光路，底部光源上的起偏器置出光路，检偏器置出光 路，落射光闸置于 C。 3、将 5X 物镜置入光路，将样品放入载物台，用粗调对标本进行对焦，微调锁定焦面，找到 观察目标，切换到高倍镜进行观察，同时调节孔径光阑适应物镜 N.A。 关机： 1、旋低载物台高度，将物镜切换到空档位或 5X 物镜位置。 2、将镜座左侧亮度调节旋钮调至最低，并关闭机身电源按钮，关闭变压器电源。 二、反射明场观察 开机： 1、确认光路：双灯室适配器拨杆成“对角线”状态；左侧三片滤光片，ND 拉出光路，NCB11 与 UV/D 置入光路；右侧起偏器及检偏器拉出光路；落射光闸置于 O 档，反射模块置于 BF 档；目镜筒右侧拉杆全部推入， 2、打开反射照明器光源，将卤素灯调节到合适亮度。 3、将 5X 物镜置入光路，将样品放入载物台，用粗调对标本进行对焦，微调锁定焦面，找到 观察目标，切换到高倍镜进行观察。 关机： 1、同明视场观察关机步骤 1；将反射照明器电压调至最低并关闭电源。 2、将反射附件左侧三滤光片、右侧两滤光片拉出光路，落射模块置于空档，光闸置 C 档。 三、荧光观察 开机： 1、确认光路：双灯室适配器拨杆正指向自己，目镜筒右侧拉杆全部推入，荧光光闸置于 C， 荧光附件至物镜间所有明视场光学附件置出光路（左侧三滤光片，右侧两滤光片）。 2、将 5X 物镜置入光路，将样品放入载物台，在明场下对焦，找到合适视野。 4、打开汞灯电源，并激发；将荧光滤色块转盘调至合适的位置，并调整光闸至“O”档。 5、利用荧光附件左侧的减光片 ND 减弱光强（系数为数值的倒数），直至荧光为合适亮度。 6、关闭明场照明，用微调锁定合适的荧光焦面，找到观察目标，切换到高倍镜进行观察。 关机： 1、将荧光滤色块转盘上的光闸调至“C”档，将荧光滤色块转盘调至空档位置。 2、旋低载物台高度，将物镜切换到空档位或 5X 物镜位置，关闭汞灯电源。 四、数码拍照 开机： 1、 打开显示器及主机电源，待系统进入桌面，硬盘灯不再频繁闪烁。 2、 打开 CCD 控制器电源，等 30 秒，待指示灯为绿色常亮。 3、 双击桌面软件图标进入 NIS 软件，点 Live，完全拉出目镜筒右侧拉杆。 关机： 1、完全退出 NIS 软件；关闭 CCD 控制器电源，然后关闭计算机，并关闭显示器电源。 注意事项： 1、 关机后需等半小时以上，待卤素灯或汞灯完全冷却方可罩上防尘罩。 2、 高压汞灯电源开机后至少间隔 30 分钟才能关闭；关闭后至少间隔 30 分钟才能开启。 3、 不用时一定要罩上防尘罩，远离灰尘、潮湿和油脂，切勿用皮肤接触任何玻璃部件。
NIKON ECLIPSE 80i 共聚焦荧光显微镜中文说明书
安全注意事项
警告
10. 安全注意事项注意。 本产品主要用于对载玻片上的细胞和组织进行显微镜观察和图像捕获。 在操作之前检查确定样品是否具有危险性。 按照您的实验室规定的标准程序操作危险样品。 如果样品具有潜在的传染性，戴上橡皮手套并避免触摸样品。如果样品与产品之间发生接触，按照您的实验室规定的标准 程序净化被污染的部分。
3
安全注意事项
警告
6. 光源发热。 灯及其周围区域（包括灯箱）在照明期间或者照明关闭后不久温度会非常高。 • 烫伤危险。在照明期间或者照明关闭后不久切勿触摸灯管或其周围区域。 • 在更换灯管之前确保灯管及其周围区域已经凉透。 • 为了避免火灾危险，切勿在灯打开时或灯关闭以后不到 30 分钟的时间内将纤维、纸或者汽油、石油醚、涂料稀释剂或 酒精等极其易燃的挥发性物质放置在灯箱附近。
8. 滤光块的处置（在产品装上数字成像头或 D-FL 落射荧光装置时）。
• 干涉滤光片（尤其是在强光照射中的激发滤）会随使用时间而退化。在用到相应的小时数后应进行更换。
• 如果滤光片曝露在湿度很大的环境中，滤光片的特性有可能发生改变。要想防止滤光片特性发生变化或退化，应避免在 湿度大或高温的环境中使用或存放滤光片。防止滤色镜遭受温度骤变。滤光片在不使用时应存放在干燥器或者带有干燥 剂的全密闭容器中。
8. 在更换滤光块时切记将灯关闭（当数字成像头或 D-FL 落射荧光装置已安装于产品上时）。 在更换滤光块时，切记关闭数字成像头或 D-FL 落射荧光装置的光源。未将灯关闭可能会使人暴露于紫外线的照射下。
9. 将 D/UV 插片留在光路中（在使用数字成像头或 D-FL 落射荧光装置时）。 在安装汞灯箱的情况下进行落射明场显微操作、落射暗场显微操作或落射 DIC 显微操作时切记将 D/UV 插片（D-ES EPI ND 插片之一）留在光路中。如果插片移出光路，紫外光将会射出，损坏您的眼睛。
荧光原位杂交技术检测自然流产中绒毛染色体异常与男女双方叶酸含量的研究
荧光原位杂交技术检测自然流产中绒毛染色体异常与男女双方叶酸含量的研究李晓泽;药泽蓉;胡志鹏;魏岳峰;梁飞;魏魏【摘要】目的利用荧光原位杂交技术(FISH)检测绒毛染色体,同时检测夫妇双方叶酸水平,寻找叶酸含量与染色体畸变二者的关系. 方法选择100例早期自然流产妇女作为病例组,100例正常早期妊娠妇女作为对照组,对病例组胚胎或绒毛运用FISH 技术进行染色体数目检测,并用绒毛染色体核型分析进行验证.检测病例组与对照组夫妇双方血清叶酸水平. 结果病例组中,FISH诊断流产绒毛染色体数目异常40例,占40.0％,以三体型最为常见,与绒毛染色体核型分析结果比较,差异有统计学意义.病例组与对照组夫妇双方叶酸水平比较,差异有统计学意义. 结论 FISH技术能够快速、准确、高效地检测自然流产或死胎染色体异常情况,血液叶酸水平低下为自然流产中绒毛染色体异常的危险因素.【期刊名称】《中国生育健康杂志》【年(卷),期】2014(025)001【总页数】3页(P75-77)【关键词】自然流产;染色体异常;荧光原位杂交;叶酸含量【作者】李晓泽;药泽蓉;胡志鹏;魏岳峰;梁飞;魏魏【作者单位】046011 山西长治长治市妇幼保健院医学遗传室;046011 山西长治长治市妇幼保健院医学遗传室;046011 山西长治长治市妇幼保健院医学遗传室;046011 山西长治长治市妇幼保健院医学遗传室;046011 山西长治长治市妇幼保健院医学遗传室;046011 山西长治长治市妇幼保健院医学遗传室【正文语种】中文荧光原位杂交技术（fluorescence in situ hybridization，FISH）是近年来兴起的分子细胞遗传学方法，被认为是诊断染色体异常的一个新技术。
本研究采用FISH 技术对100例早期自然流产绒毛细胞13、16、18、21、22、X／Y号染色体进行检测，同时检测自然流产夫妇双方的叶酸水平，结合绒毛染色体结果，探讨二者间关系。
尼康80i显微镜简单操作说明
1 开启电源：显微镜的电源开关，在显微镜后侧。
2 按下照明开关，打开透射显微观察的照明灯。
并将亮度控制旋钮调到预设位置。
3 将ND8、ND32 和NCB11 滤光块切入光路。
4 按下光路切换杆，将双目镜完成切入光路5 转动激发方式切换转盘，将其调节到无滤光块的位置上。
6 将聚光镜升到最高位置。
7 将视场光阑和孔径光阑充分打开8 将10 倍物镜移入光路。
9 放置标本后调节载物台，将标本移入视野中10 对标本对焦。
11 调节屈光度和瞳间距。
12 对聚光镜进行对焦。
13 切换到所需的物镜并观察标本。
14 观察完毕后请关闭电源。
2.4 落射荧光显微观察进行显微观察前必须：•检查照明灯的累计照明时间和落射荧光附件的状态。
如果照明灯己超过其使用寿命，请更换照明灯。
•使用无荧光型的玻片。
•使用非荧光型浸油。
•为防止标本发生褪色，请在不进行显微观察时关闭光闸。
1将明场光源关闭。
注意：A 荧光显微术用于观察荧光染色和荧光标记的样本；B 使用前明确样本荧光激发光波长和发射光波长，了解需要使用的荧光滤光片；C 荧光显微镜光学系统部件在组成以及原理上与前三种显微镜差别很大，在使用时应了解荧光显微镜光程和明场显微镜光程；D 汞灯灯室可产生高温高热，使用完请勿立即盖上防尘罩，而要在关闭电源30 分钟后再盖上防尘罩，防止烧坏显微镜部件和发生火灾E 为延长汞灯使用寿命，应按规范操作：开启汞灯变压器电源预热三分钟后再启动激发按钮，激发后就可以使用，每次使用至少开机30分钟，两次连续使用的时间间隔至少30分钟以上；2将当前所需激发方式的滤光块移入光路。
3放置标本，并调节载物台使标本进入视野。
4 切换到所需的物镜并观察标本。
5 返回明场显微观察。
转动激发方式切换转盘，将其调节到无滤光块的位置上。
拍照软件：NIS图像捕捉软件介绍打开NIS软件包，主菜单中有File Edit Camera Preprocess View Help以下主要介绍各目录下的主要子目录一．File:在此目录下主要是打开，保存，另存，Options(选项),打印，退出等其中重要的是Options点击File/options会出现一个对话框，可设置图片保存路径，前缀，格式（JPEG/TIFF等）还可设置标尺的计量单位（纳米，微米，毫米等）二．Edit：在此目录下主要是Resize(尺寸重设)，Crop（剪裁），Copy（复制）Flip Horizontally （水平翻转），垂直翻转，向左旋转，向右旋转，文件合并（File Merge）其中重要的是File Merge点击Edit／File Merge会出现一个对话框，可以分别输入红，绿，蓝三色荧光图片，点OK，就可实现三色荧光叠加。
ECLIPSE E100 显微镜 中文详细说明说
标志
警告 注意
含义 无视该标志提示有可能导致严重人身伤害甚至死亡。 无视该标志提示有可能造成人身伤害或财产损失。
产品上标志的含义
标ࢤ
ؚ义 注意！表面高温
该标志出现在视场透镜配件附近（视场透镜配件下方安装了照明灯）和底盘背面（底盘上安装了照明灯）， 提醒您注意下列情况：
警告：存在烫伤危险。照明灯及其周围区域（包括视场透镜配件）在使用时会变热。照明灯开启时或 关闭后三十分钟内，请勿与它们接触。
M413 C 07.7.STC.1
*M*M431439CENN0O11**
ECLIPSE E100 显微镜
使用说明书
感谢惠购尼康产品。 本说明书中所介绍的基本显微镜操作仅适用于使用尼康 ECLIPSE E100 显微镜的用户。 为了确保您的正确使用,请在使用前仔细阅读本使用说明书。
• 未经许可,不得翻印或转载本说明书中的任何内容。 • 本说明书的内容若有更改，恕不另行通知。 • 尼康已经尽力确保准确清晰，若您发现其中存在不清楚或不正确的内容，请就近与尼康代理商联系。 • 本说明书中提到的某些配件可能并不包含在您所购买的产品中。 • 若需将本系统与其他产品配合使用,请仔细阅读该产品的说明书。（如：显微镜、落射荧光照明灯等。） • 如未按厂商说明的方式使用本产品,可能会导致产品失效。
14. 移动固定夹
进行切换时，显微镜被牢牢固定住。 使用显微镜前，请确保固定夹己被移除。如果您想了解更详细的信息，请参阅第 27 页。
6
目录
本使用说明书中使用的警告和注意标志 ·························································································2 产品上标志的含义 ·························································································································2 通用安全说明································································································································· 3 第一章 各部件的名称························································································································8 第二章 开关和控制键······················································································································10 第三章 显微镜操作简明步骤···········································································································12 第四章 显微观察（详细步骤） ·······································································································14 第五章 其它操作·····························································································································22 1 油浸观察 ································································································································22 2 调节粗对焦螺旋的矩 ··············································································································24 3 线钩 ·······································································································································24 第六章 装配····································································································································25 1 标准套件的装配 ·····················································································································26 2 安装附加部件 ·························································································································28
尼康 TE300 Eclipse 显微镜用户指南说明书
Nikon TE300 Eclipse BrightfieldMicroscopeUser GuideLSU Health Science Center-Shreveport Research Core FacilityUser manual for Nikon Elements software Equipment: Nikon TE300 Eclipse microscopeDS Fi2 color camera (For colored images)If the room lights are not on when you arrive to use the Nikon, check and confirm that no one is using the Zeiss two-photon microscope before you turn on the light.Check whether the floating table and the microscope is clean, if not, clean it.Sign in on the sign in sheet; please use both your given name and your surname, your department, your PI, and provide a telephone number at which you can be reached. If you do not use your lab telephone as your main contact, please provide your mobile number.Turning on/preparing microscope 1. Turn on the surge protector by pressing its power button2. Turn on the computerOnce the computer is on, please login to your own user account with the following user ID and password:User ID: lsumc-master\usernameThe password is your personal school email passwordTurn on the camera before you start the software.• Turn on the Nikon Digital Sight DS-U3 unit, which is connected to the camera via a cable. Thegreen light will come on; once it stops blinking and remains a solid green, the camera is ready to use.Digital Sight DS-U3 unitDS Fi2 color cameraOpen the NIS Elements software by double clicking on the icon .A log-in dialogue box will appear after clicking on the software icon. Log in use:User ID: nikonuser12Connecting cablePassword: corescopesIn the camera dialogue box, choose the DS-U3 color camera (Neglect the other options).1. The Nikon lamp switch on the left side of the scope should be pushed in (ON ). If not, push it in.2. Choose the correct stage insert and use the nylon screws to fasten it in place. Extra screws arein a white plastic box and the yellow screwdriver is for changing stage. You can find stage inserts in the box labeled “Nikon Wide -field stage”.3. The motorized stage joystick located to the right of the microscope has three speeds: 100%, 50%and 25%, which can be changed by pressing the switch to the left of the joystick. The switchcontinuously cycles through the speeds, so try different speeds to find the one you prefer.Nikon lamp switch Screws and screwdriver for changingstage Motorized stage joystick and focus knobSetting up for imaging (objective, plates, slides)1. Lower the objective positions with the focus wheel. The lengths of the objectives are different,and the taller objectives can easily hit the stage while you turn the objective turret. 1 2 32.Place your sample securely in the stage tray. Remember to invert slides, coverslip side down.3.The microscope objectives are engineered to work best with coverslips 0.17 mm in thickness,usually referred to as number 1.5 or 1 ½ when ordering.4.Plates and dishes should not be flipped. If possible, you may want to remove plastic lids, sinceplastic can distort imaging.Observing images through the eyepieces (light path: A)1.When viewing your sample through the eyepieces, set the selector knob on the lower right sideof the scope to A, which directs 100% of the light to the eyepieces. B & D allow light to go to the eyepieces and the camera at the same time; C directs 100% of the light to the camera.Light path selector wheel2.Rotate the objective turret manually to select an objective; please do not touch the objectivesas you rotate the turret, since those with correction collars can have the collars inadvertentlychanged.3.In the OC Panel of the software, click on the objective name corresponding to the one that youhave manually selected. Note: Clicking on objectives in the OC panel does not control the actual turning of objective turret, but it tells the software which objective is currently in place andallows the software to adjust the acquisition settings best for the selected objective. Although you have to manually choose objectives, you still need to click on the corresponding objective button in the software to ensure best imaging result. Scale bars won’t be in the correct umformat if no or wrong objective is selected.[The objectives currently mounted on the turret comprise the following: 4x, 10x, 20x, 20x, 40x dry, 40x oil. Please note that only ONE (40x oil) of these is an oil objective, requiring the use of immersion fluid; the others are all air objectives, which do not require any immersion fluid, and will in fact be damaged by oil, glycerin, or water. If needed, 60x (oil) or 100x (oil) objectives are available to be checked out.]PLEASE use ONLY the Nikon oil in the brown plastic container at the microscope station with the 40X oil objective.4. Turn on the light switch5. Look through the eyepiece with transmitted light. Find and focus on your sample.Observing live images on the computer and acquisition (light path: C)1. To view samples in the software, manually set the selector knob on the lower right side of thescope to C , which directs 100% of the light to the camera.2. Click on the3. 4. Use the fine focus knob to readjust the focus on screen, because the focus through eyepiece isnot exactly the same with the focus of the software.5. Adjust the white balance of an image using the AWB button at the top tool bar of the live viewwindow.6. Capture an image by clicking the camera button .DS Fi2 (color) Camera Settings•Mode: Normal and Binning. Default Normal.•Resolution: you will see Fast (Focus) and a drop down menu, and Quality (Capture) and another drop down menu. Below those menus is a box called High Quality Capture that can be checked or unchecked.•Exposure: Uncheck auto Exposure to manually change exposure time.•Analog Gain: software sets value•AE Compensation: software sets value•Color: Auto White button will add white balance to your image.•Scene Mode >: no action•Commands: no actionSaving an image (ND, TIFF, JPEG; folders; offline access)When you are ready to save an image, go to File and choose Save As. You will have choice in which format to save the image. Each user has a personal folder located in the Users Folder on the desktop. Images can be saved there temporarily, but should be removed in good time. The easiest way is to use a flash drive and the USB ports on computer.Save your image as a .tif file from your original image. Keep in mind that TIFF format is required for publication. JPEG images is not always accepted.To save into a TIFF file. Go to File->Save as, then set the options as below:Shutting down the equipment1.If you have used oil, make sure to clean the objective with the lens paper and cleanerprovided. Currently, this only applies to the 40x oil objective used for fluorescent imaging.2.Clean your slides with cleaner and kimwipes.3.Lower the objectives and turn the objective to 4X.4.Log off and Shut Down the computer.5.Turn off the DS Fi2 color camera by turning off the Nikon Digital Sight DS-U3 unit.6.Turn OFF the surge protector under the table.7.Replace the cover and remember to sign out on the login sheet.MiscellaneousBinning averages the grey values in pixels into various sizes of bundles, which makes the signal brighter but reduces resolution. The software allows you to set the binning range for both cameras, with None as the default. The 2x2 selection will produce a good size image for focusing. If you change the binning size, you may need to redo the Auto Exposure. Using 2x2 binning, which averages the information in 4 pixels, increases your S/N ratio by a factor of 4. This is advantageous when you have a weak fluorescent signal. The use of 2x2 binning also decreases image file size and time transfer rates for the camera. Choose No Binning or the lowest value of binning for the best resolution.Tips for Better ImagingCoverslips: It is very important that your coverslip be glass, size 1 ½, 0.17 mm thick. Some of the objectives have correction collars that adjust for differences in thickness, but some do not. The correction collars may help if you are trying to image cells in plates or flasks. However, they may not be in a position ideal for your samples.Plastic: although in general images made through plastic ware are good, it may be worth the extra money to buy optically corrected plastic.Immersion medium objectives: If you are using the 40x, 60x, or 100x oil immersion objectives, please use ONLY the Nikon immersion oil next to the microscope. Residues from different oil types on the lenscan cause aberrations in the image. Kimwipes are fine for cleaning the oil off of slides (this can be followed with lens cleaner sprayed onto a Kimwipe, then used to remove the last traces of oil). When cleaning the objective lens, which is very sensitive, use the provided lens paper and draw it gently across the objective, which allows it to soak up and remove the oil without scratching or damaging the lens.For professional quality images, each component of the microscope system should be set to the optimal position for your samples before you begin to collect images. This is especially important for brightfield images that require Kohler illumination.Equipment Specifications for PublicationMicroscope: Nikon Eclipse TE300 inverted microscope with an epi-fluorescence attachmentObjectives: there are 6 objectives mounted on the turret; the rest are available for check out。
NikonECLIPSETi倒置荧光显微镜使用说明.pdf
2.打开荧光转盘上的荧光拨杆至“ 0”位置，此时荧光光路打开。旋转荧光转盘，转入所对
应的滤光片位置即可进行荧光观察； 在观察过程中可插入位于荧光灯源前方的减光片，
以改
变荧光强度，减弱对活细胞样品的损伤。
3.关机：直接关掉荧光灯电源开关即可。 注意事项：
1.短时间内频繁开关荧光灯电源，将极大的缩短荧光灯寿命。 2.荧光光源打开后即可使用，但必须使用 20 分钟才可以关闭；关闭 打开，否则会导致荧光灯损坏。
检偏器，将其放入光路后，再使用专用的银白色
DIC 物镜对焦观察。在观察过程中可旋转
起偏器，达到最佳的观察效果。
相差观察：
开机、关机等与明视场观察相同，区别是要使用聚光器相对应的相差模板 荧光观察：
PH1 。
1.开机：先打开荧光灯电源开关，然后按住松开，
Nikon ECLIPSE Ti 倒置荧光显微镜使用说明
操作步骤：
本显微镜的荧光光路与明视场（相差、 DIC 也属明视场）光路相对独立。当使用明视场
时，无需打开荧光光源， 进打开总电源即可。 当使用荧光观察时， 可同时打开总电源和荧光
电源。
明视场观察：
1. 开机：按动总电源开关“ 1”处接通电源，此时开关和指示灯点亮，打开显微镜底座左侧 的透射照明器开关，从小到大调节照明器开关下方的亮度调节旋钮，使照明亮度适当。
2. 首先将聚光器模板 A 放入光路， 用 10×物镜对样品进行对焦， 调节目镜的屈光度调节环， 然后换用 40×物镜观察样品。
3. 关机： 将亮度调节旋钮旋至亮度最暗位置， 按动总电源开关“ 0”处关闭电源。
关掉显微镜底座左侧的透射照明器开关，
然后
DIC 微分干涉观察：
开机、关机等与明视场观察相同， 区别是要使用聚光器相对应的 DIC 模板 N1 及起偏器、
激光共聚焦显微镜操作手册最终版
徐珏2011.12.3 17:59 于苏州宝石御景园目录标准探测模式（DU4）图像拍摄流程 (1)一、软件开启 (1)二、基本设置 (2)[A1 Setting]界面主要功能区域简介 (2)[Setting]界面功能简介 (4)[Filter and Dye]功能区简介 (6)串色现象解释及Line模式图解 (6)Line模式图解 (7)[scan setting]功能区简介 (7)三、获得图像 (8)[Acquisition]功能区简介 (10)放大方式选择按钮简介 (13)图片扫描参数调用简介 (17)JP2格式图片转化为JPG/TIFF格式图片的操作 (18)各通道荧光图像自由叠加的介绍 (18)数据分析 (21)一、ROI区域分析 (21)二、LUTs调节图像荧光亮度 (23)三、去背景 (25)四、标尺添加 (26)六、长度面积等常规测量 (28)七、细胞计数 (28)[Binary Toolbar]界面简介 (32)光谱扫描模式（SD模式） (34)一、SD模式基本设置 (34)[Detector]标签页功能简介 (35)[Binng/Skip]标签页简介 (36)二、图像获得 (37)三、数据分析 (39)[Spectrum Profile]视窗简介 (40)四、光谱拆分 (40)虚拟滤光扫描模式（VF模式） (47)一、VF模式基本设置 (47)[Detector]设置标签页简介 (48)[Gating Setting]标签页简介 (48)二、获得图像 (48)时间序列拍摄 (49)[Capture Timelapse]视窗简介 (49)[ND]界面简介 (51)[Time Measurement]界面简介 (53)[Capture Z-series]视窗简介 (55)拼大图拍摄 (60)光活化序列拍摄 (61)[Photo Acquition]功能区简介 (62)[Sequential Stimulation]界面简介 (63)常见细胞类型和细胞器形状 (65)常用荧光染料及应用领域 (68)标准探测模式（DU4）图像拍摄流程一、软件开启1、双击桌面NIS-Element图标。
80i荧光(操作指引)
NIKON 正置荧光显微镜80i
照相光路切换拉杆
荧光光路
视场光阑
孔径光阑 X 方向平台移动
Y 方向平台移动
色温片NCB11
减光片ND8,ND32
调焦/微调
调焦上限位
透射光视场光阑
聚光器
目镜
荧光光闸
荧光附件
荧光滤光块转盘
照相CCD
CCD 数据线
C 接口 卤素灯开关
NIKON正置荧光显微镜使用方法
一：白光观察
1.打开显微镜主机背面左侧电源开关及正前方卤素灯开关
2.调节左侧光亮度调节旋钮，把光调到合适亮度
3.把荧光滤光块转盘转到没有安装荧光滤光块的空位上。
4.把样品放在载物台上
5.使用低倍率物镜找象，调节调焦旋钮对样品对焦
6.选择观察所需的物镜对样品对焦
7.观察完毕，把亮度调节旋钮调到亮度最低，关闭正前方及背面左
侧开关。
二：荧光观察
1.打开汞灯激发器电源开关
2.把样品放在载物台上
3.转动荧光滤光块转盘，根据需要选择合适的荧光滤光块
4.打开关闸
5. 对焦观察观察，并可通过插入减光片来调节激发光的强度:注意初步找像前
把减光片都拉出来，以免荧光太弱，人眼感觉不到。
6. 观察后关闭汞灯电源开关。
注意事项：
1、确保汞灯工作30分钟以上方可关闭
2、确保汞灯关闭30分钟以上方可再次开启。
尼康D-Eclipse C1共聚焦显微镜用户指南说明书
Nikon D-Eclipse C1 Confocal MicroscopeUser GuideUniversity of Puget Sound – Department of BiologyJune 2015Table of ContentsI. Disclaimer! (2)II. Safety Considerations (2)III. Confocal Microscopy (2)IV. Operation of the Microscope (2)A. Start-up procedures (2)B. Finding your sample with conventional microscopy (3)C. Visualizing your sample with epifluorescence (4)V. Using NIS Elements to Image Epifluorescence (4)A. Adjusting Image Appearance in NIS Elements (4)B. Capturing and Saving Epifluorescent Images (6)C. Merging Captured Epifluorescent Images in NIS Elements (7)D. Stitching Multiple Epifluorescent Images together in NIS Elements (7)E. Annotating Images in NIS Elements (8)1. Add a Scale Bar (8)2. Take Measurements (10)I. Disclaimer!Please note: this is an abbreviated guide. The actual manual for this microscope + software package is easily over 300 pages. While it is a good exercise for any student to read the user manual when beginning work on a new instrument, this User Guide can serve as a guide to general use of the instrument and may be sufficient for many uses. However, in the event that you encounter trouble, need more information, or require assistance, contact Amy (email: ************************;location:Room117H,ThompsonHall;phone:253-879-2829)orrefer to the manual for the instrument or software.II. Safety ConsiderationsThe confocal microscope utilizes high powered lasers which emit intense beams of light. Though risk is minimal since the system is enclosed, some hazards remain. Exposure to lasers may result in damage to the eyes or skin, among other hazards. For these reasons, please use the following precautions when working on the confocal microscope:∙Never look directly into any laser beam light source.∙Adhere to all safety warning labels found on the microscope and lasers.∙Notify a supervisor, faculty member, or other responsible individual immediately if a laser is malfunctioning or losing power.For additional information on laser safety associated with using the confocal microscope, consult the Nikon website at /articles/fluorescence/lasersafety.html.III. Confocal MicroscopyConfocal microscopy is an imaging technique commonly used in the biomedical and life sciences to visualize fixed or living cells and tissues, among other applications. Confocal imaging is based on principles invented by Marvin Minsky in 1957, and involves the use of point illumination and elimination of light transmitted from the out-of-focus planes via a pinhole located in front of the detector. This imaging technique provides a significant increase in optical resolution and contrast over traditional wide-field fluorescent microscopy, and thus allows for the resolution of fine structure, acquisition of 3D images, and achievement of exquisitely-fine focus.IV. Operation of the MicroscopeStudents should always have training on the microscope prior to use! To arrange to be trained, or forotherassistance,pleaseseeAmyin117HThompsonHall(****************************** 879-2829).A. Start-up proceduresThe following sequence should be used when turning on the microscope (summary located on the wall next to the microscope):1. Sign up on the online instrument calendar located on the science core facility webpage:/sciencecorefacility.2. Turn on 2) the power strip for the lasers and the epifluorescence camera. This is located in thecorner on the bench to the left of the microscope.3. Turn on 3) each laser individually, only if you are planning to do confocal microscopy. These DONOT need to be turned on if you are performing epifluorescence. Ideally, the lasers should be onfor 20-30 minutes prior to use. The time that it takes to turn everything on and set up the microscope is generally enough time for the lasers to warm up properly. However, in the case of quantitative analysis of multiple samples, be sure to wait for an appropriate amount of time (ie, 20-30 minutes) from start-up to begin collecting images and data.a. 405 nm laser: Silver control box on the back of main lasers box. Turn the key located onthe back of the laser. Key in the horizontal (o) position is off. Key in the vertical (l) positionis on.b. 488 nm laser:i. Turn the key located on the black controller box to the left of the main lasers box(key in the vertical position (o) is off, key in the horizontal position (l) is on);ii. When green light stops flashing, push down the green “laser on” button.c. 561 nm laser: Silver control box to the front of the main control lasers box. Switch in thevertical (o) position is off. Switch in the horizontal (l) position is on.4. Turn on 4) the powerstrip by the computer. This controls power to the controller and detector.5. Turn on 5) the computer and monitor.6. Open the appropriate software for imaging. EZ-C1 for confocal laser scanning or NIS Elements forepifluorescence.B. Finding your sample with conventional microscopy1. Make sure the lowest objective is in place, and stage is lowered with the course focus to reducethe risk of running the objective into the stage or slide.2. Place the slide, coverslip-down, on the platform (remember, this is an inverted microscope, so theobjective is underneath the sample).3. Be sure the port knobs (located on the right side of the microscope) is pointing to the indicator for“EYE”, the shutter is open (lever located just behind the filter turret on the right side of the microscope, pushed back to the “O” position), and the filter turret (located ju st below the objectives) is in the transmitted light position “ ”, which is used for bright field imaging to initially find your specimen before switching to fluorescence.4. Turn on the transmitted light by pressing the white button on the lower left-hand side of themicroscope. You can adjust the intensity by turning the dial just below the white button.5. Using the eyepieces, find your cells/tissues/etc., and focus using the coarse focus dial or theremote fine focus controller dial. The platform can be moved to reposition your specimen using the rod on the right side of the microscope, rather than actually manually moving your slide.Note: Depending on your tissue, it might be hard to see with bright field microscopy. If that is the case, you might find it helpful to focus on the edge of your coverslip to get you close to the plane of focus, then position where you think your sample is over the objective and continue on to the next section “Visualizing your sample with epifluorescence”.6. Start with the 10x objective, find your cells/tissue/etc., and then switch to the 20x, 60x, or 100xobjective if needed. If using the 60x WI objective, place a drop of double distilled DI water on the top of the objective before use. If using the 100x Oil objective, place a small drop of immersion oil on your slide.Note: The 100x objective is kept the shelf to the left of the microscope. You must be trained prior to use of the 100X objective. Please use it carefully, be sure to clean itcorrectly after use, and take care when mounting and unmounting it. If you need assistance, please contact Amy.C. Visualizing your sample with epifluorescence1. Once you have found and focused on your sample using bright field microscopy, turn off thetransmitted light.2. Place the filter turret (located just below the objectives) to the correct position for the fluorescenceyou wish to view. Filters are labeled as follows:∙“UV”: UV excitation, used to visualize DAPI∙“B”: blue excitation, used to visualize GFP and FITC∙“G”: green excitation, used to visualize Cy3, TRITC3. Turn the LED light source on by tapping the foot pedal located on the floor underneath the tablethe microscope is sitting on. Note: Be sure to turn off the LED light source by tapping the foot pedal again w hen you don’t need to visualize fluorescence or if you have switched to confocal microscopy. You never want your sample exposed to unnecessary light as this might quench your fluorophore, resulting in reduced fluorescence.V. Using NIS Elements to Image EpifluorescenceA. Adjusting Image Appearance in NIS Elements1. Switch to the computer view: Turn the top portal knob (on the right side of the microscope, acrossfrom the CoolSnapEZ camera) to “SIDE”.2. Open the NIS Elements software by double-clicking the icon on the desktop, select RoperScientific and click OK.3. To view the live image, click the button in the upper left hand corner of the software. You willalso need to indicate what filter you are currently using for your image to appear the correct color.There are several presets in the tool bar, one for each emission color (see image below). Once you choose the correct fluorophore the tab at the lower left corner of your live image will display the correct label (see image below).4. If you need to change the label, or if the presets have been changed, right click the tab circled inred above and choose Properties. In the Name drop-down menu you can choose from a variety of fluorophores. Sometimes you may also need to change the color of those presets. Click Close when finished.5. To adjust the brightness of the image you will first want to select “Auto Exposure” from theCoolSNAP ES Settings on the right-hand side of the screen.6. To adjust the appearance of your image you can adjust the LUTs (Lookup Tables) located on theright-hand side of the screen.∙During the imaging process, the camera collects the transmitted light and assigns a value to each pixel based on its intensity. Brighter pixels are assigned higher intensity values. The intensity values are arranged in histograms along the x-axis, with lower intensity values on the left and higher intensity values on the right. The height of the bar at a particular intensity value represents its pixel population. Higher bars indicate more pixels at that intensity value.∙How these intensity values are assigned, or ‘mapped’, greatly affects the visual appearance of the image, although it does not affect the data or quantification in any way. The curve overlaying the histogram represents how the intensity values of the data map to the available display values. Adjusting the curve changes which of the intensity values are mapped to specific display values, and whether they are mapped linearly or logarithmically.∙There are three points on the curve that can be adjusted:a. The Max Point is at the right of the curve. Moving the Max Point to the left is similar toincreasing the Contrast. As more of the intensity values are mapped to the brightestdisplay values, fluorescence becomes brighter.b. The Min Point is at the left of the curves and marks the lowest display value that is not inthe background color. Moving the Min Point to the right shades more of the lower intensityvalues to lower display values, creating a visually cleaner background.c. The middle point represents the K value. Adjusting it smoothly changes the mapping fromlinear to logarithmic. Changing to a more logarithmic mapping reduces the contrastbetween the higher and lower intensity values by applying a log scale, so higher and lowerintensity values are mapped to display values that are closer together. This ‘compression’of the intensity values may improve the appearance of the less intense bands, whileavoiding very bright bands.Min PointB. Capturing and Saving Epifluorescent Images1. To capture the image click the camera icon in the tool bar. This will open a new window with yourcaptured image.2. To save the image, go to “File” then choose “Save As” and find your folder. Note all students musthave a subfolder within their adviser’s folder.3. Give your file a name and select what kind of file type you’d like to save your image as. There arethree recommendations. NOTE: If you save your image in TIFF or JPEG formats you must include the magnification in your image name or write it down in your lab notebook in order to go back and calibrate the image to the correct scale bar or make measurements.a. ND2 Image File Format. This saves your image in the proprietary NIS Elements formatsthat allows you to keep information about the instrumentation such as instrument type andmagnification with your image. This is particularly helpful if you don’t have a file namingconvention that includes the magnification your image was acquired at. You can processyour image in this file format and then save it in one of the following formats without therisk of losing image quality.b. TIFF, Tagged Image Format. This is a powerful image format that is the standard forgraphic artists, publishing, and photography. TIFFs are a lossless formatting meaning thata TIFF image may be edited and re-saved without losing image quality. One disadvantageto TIFFs is that they can’t always be viewed on all devices.c. JPEG, Tagged Image Format. This is a particularly useful file type that can be viewed onvirtually any device. The disadvantage to using JPEGs is that image quality may be lostwhen editing and resaving your image.C. Merging Captured Epifluorescent Images in NIS ElementsIf you have a sample with more than one fluorophore you can merge together two fluorescent images and/or merge a fluorescent image with a brightfield image.1. Go to the “File” tab then click “Merge Images…”.2. In the “Merge Images” window that appears you will select the file name of the images you want tomerge from the drop down menu under the appropriately colored component or Brightfield option, then click “OK”.3. Save the image according to the instructions in section B above. If you need to annotate your imagewith text, arrows, scale bar, or make measurements see section 5E below.D. Stitching Multiple Epifluorescent Images together in NIS ElementsIf you have a large sample or piece of tissue you want to see all together at once you can capture multiple images and then stitch them together.1. Capture your images in a systematic way by always moving across in the same direction or down ina column. Also make sure you capture the image for all fluorophores or under brightfield beforemoving on to the next area. Note: Images must be saved in a file format other than .ND2, such as JPEG or TIFF in order to stitch them together.2. Go to the “File” tab then click “Stitch Larger Image from Files…”, then open your image sequenceby navigating to the folder for the images that you want to stitch together.3. Select the correct layout options so that your image is stitched together in the correct order. It’shelpful if you remember the direction you moved to obtain images and how many columns you captured. Click Stitch.E. Annotating Images in NIS Elements1. Scale Barsa. Adding a scale bari. On the right hand side of your image there will be a tool bar. To add a scale bar,select the icon that that has “µm” on it.ii. If you are adding it to a .ND2 file, a calibrated scale will be added and you canproceed with editing the scale bar properties. If you are adding it to any other filetype you will need to calibrate the scale bar from pixels to µm.iii. To calibrate your scale you will need to right click on “Uncalibrated” which is locatedon the bottom of your image window to bring up the calibration menu.iv. Select “Calibrate using Objective” and then click on the appropriate objective. Youshould not have a scale bar on your image (see example below) and proceed with editing scale bar properties.b. Edit scale bar propertiesi. To change the appearance of your scale bar, click on the small arrow to the right ofthe “µm” icon and select “Scale Properties…”ii. In the new window that opens you can select the orientation, appearance, size, and font of your scale bar, then click “OK”.c. Saving an image with a scale barIn NIS Elements, annotations act like layers (or overlays) in an image. In order to have the scale bar appear on your saved image you first need to insert the overlay by going to the “Edit” tab, then select “Insert Overlay”. You can now save the image according to theinstructions in section 5B above. Note: It is recommended to save your annotated image as a new file name so you always have your original image to go back to.2. Take Measurementsa. Annotations and Measurements toolboxi. There are a number of measurements that the NIS Elements software can do with acalibrated (correctly scaled image) that a pretty self-explanatory once you find thetools. The “Annotations and Measurements” toolbox can be found in the lower lefthand side of the screen beneath the “CoolSNAP ES Settings” and “LUTs” toolboxes.Sometimes this toolbox accidently gets closed. To view it again go to the “Measure”tab at the top of the screen and select the measurement you would like to make.ii. One nice feature of the “Annotations and Measurements” toolbox is that it will keep track of any measurements or counts that you have made in an Excel Spreadsheetacross a set of pictures. This spreadsheet can be exported and saved.b. Saving an image with annotationsRemember: In NIS Elements, annotations act like layers (or overlays) in an image. In order to have the scale bar appear on your saved image you first need to insert the overlay bygoing to the “Edit” tab, then select “Insert Overlay”. You can now save the image according to the instructions in section 5B above. Note: It is recommended to save your annotated image as a new file name so you always have your original image to go back to.。
NIKON 80i中文说明书
1. 在组装、连接/断开电源线、换灯和维护时，将产品电源断开。 为了防止触电和/或故障，在组装、连接/断开电源线、换灯和清洁产品及物镜时，切记关闭产品的电源开关（按下开关， 至“!”位置）并从仪体插座上拔下电源线。
2. 更换灯注意事项。 为防止烫伤，在关灯后至少等待 30 分钟，使其有足够的时间冷却。为了避免触电或故障，切记在换灯之前先将产品电源 开关和外围设备开关关闭（按下电源开关按钮令其弹出），然后从壁仪体插座拔下电源线。 换灯以后确保灯箱盖牢牢固定到灯箱。切勿在灯箱盖打开时开灯。切勿打碎用过的灯，相反，将其作为特殊工业垃圾或按 照当地法规的规定进行处置。
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产品搬运处置说明
产品搬运处置说明
1. 搬运产品时动作要轻。 本产品属精密光学仪器，搬运时动作要轻。避免使其受到突然的冲击和振动。. 即便是相对较弱的冲击力也有可能影响物镜的精密度。
2. 弱电磁波。 产品会发出微弱的电磁波。切勿将精密电子仪器放置在产品近旁，以免降低其性能。如果电视或无线电接收受效果到影响， 将电视或无线电移至远离产品的地方。
4. 显微镜的电源线和交流适配器的电源线。 使用规定的电源线中的一种。误用电源线可能会导致起火或其它危险。 该产品属于一级触电保护产品。一定要将其连接到适当的接地端子。 关于规定的电源线，参阅第 8 章。 了防止触电，在拔插电源线之前一定要关闭产品总电源开关（按它至“0”位）。
5. 安全注意事项警告 检查交流适配器（在使用 D-CBC-BOX 集线箱时）D-CB C-BOX 只适用规定的型号交流(AC)适配器供应用。使用任一种其他型 号适配器会导致故障，过热，甚至失火。参阅第 8 章适配器规定。 • 防止故障和失火交流适配器应放通风良好处，为防止热辐射及过热，切勿遮盖和防止东西在适配器上面。 • 为防止故障，在接近交流适配器之前，总是切断 C-BOX 的开关。
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安全注意事项
警告
6. 光源发热。 灯及其周围区域（包括灯箱）在照明期间或者照明关闭后不久温度会非常高。 • 烫伤危险。在照明期间或者照明关闭后不久切勿触摸灯管或其周围区域。 • 在更换灯管之前确保灯管及其周围区域已经凉透。 • 为了避免火灾危险，切勿在灯打开时或灯关闭以后不到 30 分钟的时间内将纤维、纸或者汽油、石油醚、涂料稀释剂或 酒精等极其易燃的挥发性物质放置在灯箱附近。
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产品搬运处置说明
6. 调焦手轮。 • 切勿同时反向旋转产品左右两边的调焦手轮。这样做有可能会损坏产品。 • 旋转粗调焦手轮超过它的调节最大范围，可能对产品产生损害。在调节手轮时请不要过分用力。
7. 防止灰尘和外来光进入端口（在三目镜筒、数字成像头或 C-TE 人机学双目镜筒装好后）。 为了阻挡外来光和灰尘，切记在不使用的端口时盖上配置的盖子。
3. 使用专用的电源装置、灯和灯箱。 切记使用专用的电源装置、灯和灯箱。使用非专用的电源装置、灯和灯箱有可能导致故障或起火。关于专用的电源装置、 灯和灯箱，请参阅第 8 章。
4. 避免与水接触。 切勿使水与产品接触，把产品放置在远离液体的地方。水溅入产品后有可能导致短路，致使发生故障或异常发热。如果水 溅到产品上面，应立即关闭电源开关（按压它至“！”位置）并从插座中拔下电源线。然后用干布或类似物品揩干水分。 如果水进入产品，切勿继续使用；出现这种情况，请您就近与分销商联系。
8. 滤光块的处置（在产品装上数字成像头或 D-FL 落射荧光装置时）。
• 干涉滤光片（尤其是在强光照射中的激发滤）会随使用时间而退化。在用到相应的小时数后应进行更换。
• 如果滤光片曝露在湿度很大的环境中，滤光片的特性有可能发生改变。要想防止滤光片特性发生变化或退化，应避免在 湿度大或高温的环境中使用或存放滤光片。防止滤色镜遭受温度骤变。滤光片在不使用时应存放在干燥器或者带有干燥 剂的全密闭容器中。
7. 汞灯和氙灯（当数字成像头或 D-FL 落射荧光装置已经安装成）。 用于数字成像头或 D-FL 落射荧光装置的汞灯（或氙灯）在操作时需要特别注意。为了系统的安全和正确使用，请仔细阅 读下面的警告。牢记所有可能的危险。另外，仔细阅读超高压汞灯电源（或高亮度光源）的说明书和灯管生产商的说明书 （如有），然后遵守这些说明书的指示。 汞灯和氙灯的危险 1) 汞灯（和氙灯）在点亮时放射出紫外光，能够损坏您的眼睛和皮肤。直接观察灯光有可能致盲。 2) 灯管含有压力非常高的密闭气体，打开灯时压力会升高。如果灯管被擦伤、污染，受到很高的外部压力或物理冲击力， 或者超寿命使用，密闭气体有可能逸出，或者灯管有可能爆裂，导致气体吸入、玻璃伤人或其它伤害。 3) 当灯开亮时，灯及其周围区域会变得非常热。直接用手触摸灯有可能导致烫伤；放置在灯附近的易燃物质有可能起火燃 烧。 4) 用错灯的型号有可能导致发生事故，包括灯管爆裂。 安全是尼康产品的首要设计考虑因素。只要用户遵守本说明书列出的警告和注意事项，指本系统用于设计用途，那么前述 危险不会真正发生。 相反，如果不注意本说明书列明的警告和注意事项，使本系统受到振动或冲击，或者试图拆卸本系统，则有可能导致事故 和伤害。务必熟悉并遵守所有警告和注意事项。
M318C 05.1.STC.1(1/2)
显微镜 ECLIPSE 80i
说明书 <显微术>
导言
感谢惠购尼康产品。 本说明书是对显微镜基本操作的说明，供尼康 ECLIPSE 80i 显微镜的用户使用。. 为了确保正确使用，请在操作产品之前仔细阅读本说明书。
• 未经尼康公司明确许可，不得翻印或转载本说明书的全部或部分内容。 • 本说明书的内容若有更改，恕不另行通知。 • 尽管我们已尽最大的努力确保本说明书的准确性，但是，仍有可能存在讹误或不一致的地方。如果发现不清楚或不正确之
3. 仔细阅读各类说明书。 为了确保安全，仔细阅读本说明书以及与本产品配套使用的其它设备的说明书。注意每一本说明书开头部分列出的警告和 注意事项。 当数字成像头或 D-FL 落射荧光装置已经安装到产品时： 落射荧光显微术使用的汞灯（或氙灯）在操作时需要特别注意。请务必先阅读光源（超高压汞灯电源或高亮度光源）的说 明书。
5. 切勿将任何物品放置在产品顶上。 切勿将任何物品放置在产品顶上，或者用布或类似物品盖住本产品。这样做会使本系统温度上升，导致故障。
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安全注意事项
注意
6. 产品组装和安装的注意事项 • 组装产品时小心夹着您的手或手指。 • 光学元件（如透镜和滤色镜）上的擦痕或指痕会使显微镜图像的质量下降。谨防留下擦痕或者与透镜和滤色镜直接接 触。 • 产品主体部分约重 12kg。使用位于镜臂前部和主体部分底部的凹进处抓握主体部分。 • 在提带显微镜之前从显微镜上拆下所有附件（如已安装）。 • 切勿将产品安放在橱柜中。
5. 安装地点。 本产品属精密仪器。在不当环境中使用或存放有可能导致故障或性能不佳。在选择安装地点时请考虑以下因素： • 选择没有振动的地点。将产品安装在水平的表面上。 • 将显微镜和电源装置安装在离墙至少 10cm 以外的地方。 • 选择一旦发生碰撞、地震或其它潜在灾难时危险较小的地点。为了防止产品掉落，必要时使用结实的绳索或其它方法 将其固定到工作台或重量大而稳定的其它物体上。主体部分（背面）底座凹进处后面有 2 个 M6 螺孔（左右各一个）， 可以用于固定产品。 • 避免放置在太阳光直射的地点、室内光线直射的地点和其它明亮的地点。 • 避免灰尘过多的地点。 • 为了避免液体泼溅，不要在近水的地方使用产品。 • 确保环境温度在 0-40℃之间，湿度在 85%（含）以下。将产品安装在热而潮湿的地点有可能导致发霉或凝露，从而 影响性能或者引发故障。 • 切勿将产品安装在橱柜中。 • 选择出现紧急情况时便于将电源线从电源装置的交流插座中拔出的布局。 • 位于产品正上方的室内灯光有可能射入物镜，成为外来光。如有可能，在进行观察时关掉位于产品正上方的室内灯光。 • 切勿将显微镜放置在桌垫或类似的垫子上面。
处，请您就近与尼康公司的代表联系。
• 本说明书提供说明的部分产品可能不包含在您所购买的产品之中。 • 对于本产品的附件或配套使用的其它产品（超高压汞灯电源、高亮度光源等），请您务必阅读其相关说明书。
本说明书使用的警告/注意符号
尽管尼康产品的设计提供了最高的安全性，但是，忽视安全注意事项或不当使用仍有可能导致人身伤害或财产损坏，还有可能使质保条款 失效。为了确保安全使用，在操作设备之前请仔细完整地阅读本说明书。切勿丢弃本说明书。请将其保管在产品近旁方便存取的位置，随 时备考。 在本说明书中，安全提示下列符号标示。为了产品的安全和正确使用，切记遵照标有这些符号的说明。
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产品搬运处置说明
产品搬运处置说明
1. 搬运产品时动作要轻。 本产品属精密光学仪器，搬运时动作要轻。避免使其受到突然的冲击和振动。. 即便是相对较弱的冲击力也有可能影响物镜的精密度。
2. 弱电磁波。 产品会发出微弱的电磁波。切勿将精密电子仪器放置在产品近旁，以免降低其性能。如果电视或无线电接收受效果到影响， 将电视或无线电移至远离产品的地方。
符号
提示
警告
无视标有该符号的提示可能会导致死亡或能会导致受伤或财产损坏。
1
导言
本产品所使用符号的含义
如果产品标有以下符号，表示在使用过程的任何时候都需注意。 在使用或调节标有符号的任何部件之前，请查询说明书并阅读相关的说明。
注意！生物危害 在载物台上发现这一符号表示以下含义： • 警告：样品与产品的接触可能导致发生生物危害的危险。 • 为了防止生物危害的污染，避免直接用手触摸被污染部分。 • 按照您的实验室规定的标准程序净化被污染的部件。 注意防热 ECLIPSE 80i 灯箱上的这一符号表示以下含义： • 灯及其周围区域（包括灯箱）在照明期间或者照明关闭后不久温度会非常高。 • 烫伤危险。在照明期间或者照明关闭后不久切勿触摸灯管或其周围区域。 • 在更换灯管之前确保灯管及其周围区域已经凉透。
4. 显微镜的电源线和交流适配器的电源线。 使用规定的电源线中的一种。误用电源线可能会导致起火或其它危险。 该产品属于一级触电保护产品。一定要将其连接到适当的接地端子。 关于规定的电源线，参阅第 8 章。 了防止触电，在拔插电源线之前一定要关闭产品总电源开关（按它至“0”位）。
5. 安全注意事项警告 检查交流适配器（在使用 D-CBC-BOX 集线箱时）D-CB C-BOX 只适用规定的型号交流(AC)适配器供应用。使用任一种其他型 号适配器会导致故障，过热，甚至失火。参阅第 8 章适配器规定。 • 防止故障和失火交流适配器应放通风良好处，为防止热辐射及过热，切勿遮盖和防止东西在适配器上面。 • 为防止故障，在接近交流适配器之前，总是切断 C-BOX 的开关。
7. 在接通电源之前除去产品的所有遮盖物。 在使用产品时切勿用布等物品进行遮盖，因为这样会使温度异常升高，有可能导致起火。.
8. 长时间持续观察的注意事项。 为了减轻长时间观察导致的疲劳，将持续观察的时间限制在 1 小时。持续观察之间至少休息 10-15 分钟。调整其它设备的 布局及椅子的高度。
9. 产品处置。 为了避免发生生物危险，按照您的实验室规定的标准程序将产品作为被污染设备进行处置。
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安安安安安项
请遵守以下安全注意事项。
警告
1. 产品的设计用途。 本产品通过透射和落射照明，主要用于对切片上的细胞和组织进行显微镜观察和图像捕获。 其设计用途是在医院和其它实验室设施的病理学和细胞学领域对细胞和组织进行实验和观察。