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M系列调制叶绿素荧光成像系统IMAGING-PAM

简要描述:全叶片荧光成像,精确到每个像素,光合作用横向异质性分析,多达17个荧光参数,测量荧光诱导、暗弛豫、光响应曲线等,代表了调制叶绿素荧光技术的未来发展趋势。

  • 更新日期:2024-01-30
  • 访  问  量: 10513

详细介绍


传统的光纤型调制荧光仪(如pam-2100、mini-pam等)只能测量叶片上一个点的光合活性。利用一个点的数据代表一个叶片,利用一个叶片代表一个植株,进而代表一个群体(如森林、大田作物等),这种方法的误差是比较大的。

从1980年代末期开始,科研人员就期望能通过成像型荧光仪来测量全叶片光合活性,并进行了不懈的尝试,但受技术上的限制,所设计的仪器无法商品化或商品化了但得不到大家认可。其中一个很重要的原因就是能够发出饱和脉冲水平强光的二极管(led)尚未面世。要利用调制荧光技术测量全叶片水平的光合作用,首先要保证叶片上任何一点所接受到的光强必须是*相同的。调制荧光技术要求光源必须能发出很强的饱和脉冲光。卤素灯能发出很强的光,但其光场非常不均匀,根本不能用于成像!装在一个平面上的led阵列发出的光很均匀,但在2000年前,能发出*光合有效辐射(par)的led根本没有面世!

2000年,能发出*par的蓝光led面世。2001年,zui的调制荧光仪制造商德国walz公司设计制造了真正的*台多功能调制荧光成像系统imaging-pamimaging-pam采用*发光led作为光源,保证叶片表明受光均匀且光强足够强;imaging-pam采用ccd作为检测器,能检测叶片上每个像素的光合作用;imaging-pam秉承了walz公司pam系列荧光仪的一贯优点,功能强大,测量参数多,操作极其简单,一面世就受到植物学家的青睐,迅速占领市场

2005年,walz又推出了m系列imaging-pam,一个主机可以连接不同的探头(microscopy-,micro-,mini-和maxi-探头),分别在130×150 um、3.5×4.5 mm、24×32 mm或10×13 cm的面积上测量荧光成像。现在,只需一个主机连接不同的探头,即可满足从单细胞到全叶片,从分子生物学到生态学研究的全面需要

m系列imaging-pam不同版本的比较

maxi-版

mini-版

micro-版

microscopy-版

成像面积10×13 cm

成像面积24×32 mm

成像面积3.5×4.5 mm

成像面积130×150 um

放大1.5

放大6

放大45

放大130-1300


功 能
* 一个主机连接不同的探头可满足从单细胞到全叶片、从分子生物学到生态学的不同需求
* 全叶片光合作用分析(荧光成像),可测荧光诱导曲线并进行淬灭分析
* 可测快速光响应曲线(120 s内完成,比光合放氧和气体交换等技术快得多)
* 叶片光合作用的横向异质性检测
* *相同的条件下同时测量多个样品(植物、地衣、苔藓、微藻等)
* 遗传育种、突变株筛选的强大工具
* 不同的测量面积,不同的分辨率
* 可利用多孔板(如96孔板)做多个微藻样品的同时成像
* 胁迫损伤的早期检测
* 不连接显微镜即可测量绿色荧光蛋白(gfp)荧光
* 可测量叶片吸光系数


测量参数


* 以上所有参数均可成像
* 吸光系数abs和新参数ql、y(npq)和y(no)的成像是imaging-pam*的
* 生态毒理学研究中,选一个参考点,可以直接求出其它处理(如农药)的受抑制程度inh.



各种荧光参数的成像是将0.0(黑色)至1.0(紫色)的数值转换成颜色来显示的。


应用范围

* 环境科学
* 水生生物学
* 海洋与湖沼学
* 生态毒理学
* 园艺学
* 农业科学
* 林学
* 环境科学
* 水生生物学
* 海洋与湖沼学
* 生态毒理学
* 园艺学
* 农业科学
* 林学


dcmu在叶片中的渗透过程


m系列imaging-pam不同版本介绍

maxi-版
大探头,成像面积10×13 cm

调制荧光成像系统的maxi-探头利用300 w的led阵列,可以在10×13 cm的面积上提供均为的调制测量光、光化光和饱和脉冲光。该探头的支架上配备特制护眼遮光罩,可以在保护眼睛的同时观测到红色荧光的变化。

walz提供两种数码相机ccd供选择。用户若需要高清晰度,推荐选择imag-max/k[2/3" chip, 1392×1040象素, 4象素组合(binning)技术]。标准应用可选择imag-max/k2(1/2", 640×480象素),可与imag-max/k2z物镜(f1.0/f=8-48mm)结合使用。



测量盆栽植物

测量离体叶片

测量微藻样品

新增镜头可调放大倍数

y(npq)

ps/50

f

96个微藻样品成像



mini-版
小探头,成像面积24×32 mm

调制荧光成像系统的mini-探头采用强大的luxeon led阵列,包括4组(每组3个)led,均配有长波截止滤光片。配备8个红光(650 nm)和8个近红外(780 nm)led,用于测量叶片吸光系数的成像。

3种版本可选
imag-min/b:蓝光,450 nm,测量叶片等;
imag-min/r:红光,620 nm,测量蓝藻
imag-min/gfp:蓝光,480 nm,测量绿色荧光蛋白(gfp)

由于mini-探头的便携式设计,使其特别适合野外应用。由于mini-探头的成像面积仅为maxi-探头的1/16,因而前者发出的zui大光强更大,但耗电却小得多。mini-探头可以安装在光合仪gfs-3000的叶室3010-s上,同步测量全叶片气体交换和荧光成像,并且其光源可由gfs-3000控制,达到真正的同步测量
mini-探头采用1/3"数码相机ccd(640×480象素)和f1.2/f=12mm物镜。其设计目的为测量固定距离下的荧光成像。

与光合仪gfs-3000连用

长时间测量可装在三角架上

fm

qn



micro-版
微探头,成像面积3.5×4.5 mm

调制荧光成像系统的micro-探头是一个极便携的探头,采用整合式cosmicar-pentax cctv物镜(f1.4/f=16mm),直接安装在数码ccd(1/3" chip, 640×480像素)上。

micro-探头只配备一个luxeon led(蓝光,450 nm)和一个特制双色分光镜,类似于落射荧光显微镜。

尽管成像面积只有3.5×4.5 mm,但45倍的放大率却允许对叶片荧光成像的异质性分析达到支脉(minor veins)级。同时还可提供一个特制版本用于测量gfp的成像。


micro-探头还可安装在标准版imaging-pam(2001年设计)主机上。该探头提供x-y轴可调的样品台。其设计目的为测量固定距离下的荧光成像。

fo

gfp成像

微探头直接安装在ccd上



microscopy-版
显微探头,成像面积130×150 um

必须与特制落射荧光显微镜(hund或zeiss)结合使用,该显微镜可以提供激发光并检测荧光

imag-max/k(数码相机ccd)[1392×1040象素,4象素组合(binning)技术]可以提供高灵敏度。

探头标准配置是一个*luxeon led(450-480 nm),用于提供测量光、光化光和饱和脉冲。目前已可提供rgb探头(红-绿-蓝-白led光源),它采用了phyto-pam技术,可以显微镜下自动对蓝藻、绿藻、硅/甲藻、红藻进行分类并测量光合作用。


分类,红色为硅藻,绿色为丝状绿藻

光合,fv/fm活性,可区分细胞不同部位的活性


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