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傳統(tǒng)的光纖型調(diào)制熒光儀（如pam-2100、mini-pam等）只能測量葉片上一個(gè)點(diǎn)的光合活性。利用一個(gè)點(diǎn)的數(shù)據(jù)代表一個(gè)葉片，利用一個(gè)葉片代表一個(gè)植株，進(jìn)而代表一個(gè)群體（如森林、大田作物等），這種方法的誤差是比較大的。

從1980年代末期開始，科研人員就期望能通過成像型熒光儀來測量全葉片光合活性，并進(jìn)行了不懈的嘗試，但受技術(shù)上的限制，所設(shè)計(jì)的儀器無法商品化或商品化了但得不到大家認(rèn)可。其中一個(gè)很重要的原因就是能夠發(fā)出飽和脈沖水平強(qiáng)光的二極管（led）尚未面世。要利用調(diào)制熒光技術(shù)測量全葉片水平的光合作用，首先要保證葉片上任何一點(diǎn)所接受到的光強(qiáng)必須是*相同的。調(diào)制熒光技術(shù)要求光源必須能發(fā)出很強(qiáng)的飽和脈沖光。鹵素?zé)裟馨l(fā)出很強(qiáng)的光，但其光場非常不均勻，根本不能用于成像！裝在一個(gè)平面上的led陣列發(fā)出的光很均勻，但在2000年前，能發(fā)出*光合有效輻射（par）的led根本沒有面世！

2000年，能發(fā)出*par的藍(lán)光led面世。2001年，zui的調(diào)制熒光儀制造商德國walz公司設(shè)計(jì)制造了真正的*臺(tái)多功能調(diào)制熒光成像系統(tǒng)imaging-pam。imaging-pam采用*發(fā)光led作為光源，保證葉片表明受光均勻且光強(qiáng)足夠強(qiáng)；imaging-pam采用ccd作為檢測器，能檢測葉片上每個(gè)像素的光合作用；imaging-pam秉承了walz公司pam系列熒光儀的一貫優(yōu)點(diǎn)，功能強(qiáng)大，測量參數(shù)多，操作極其簡單，一面世就受到植物學(xué)家的青睞，迅速占領(lǐng)市場。

2005年，walz又推出了m系列imaging-pam，一個(gè)主機(jī)可以連接不同的探頭（microscopy-，micro-，mini-和maxi-探頭），分別在130×150 um、3.5×4.5 mm、24×32 mm或10×13 cm的面積上測量熒光成像?，F(xiàn)在，只需一個(gè)主機(jī)連接不同的探頭，即可滿足從單細(xì)胞到全葉片，從分子生物學(xué)到生態(tài)學(xué)研究的全面需要。

m系列imaging-pam不同版本的比較

maxi-版	mini-版	micro-版	microscopy-版





成像面積10×13 cm	成像面積24×32 mm	成像面積3.5×4.5 mm	成像面積130×150 um

放大1.5倍	放大6倍	放大45倍	放大130-1300倍

功能
＊一個(gè)主機(jī)連接不同的探頭可滿足從單細(xì)胞到全葉片、從分子生物學(xué)到生態(tài)學(xué)的不同需求
＊全葉片光合作用分析（熒光成像），可測熒光誘導(dǎo)曲線并進(jìn)行淬滅分析
＊可測快速光響應(yīng)曲線（120 s內(nèi)完成，比光合放氧和氣體交換等技術(shù)快得多）
＊葉片光合作用的橫向異質(zhì)性檢測
＊ *相同的條件下同時(shí)測量多個(gè)樣品（植物、地衣、苔蘚、微藻等）
＊遺傳育種、突變株篩選的強(qiáng)大工具
＊不同的測量面積，不同的分辨率
＊可利用多孔板（如96孔板）做多個(gè)微藻樣品的同時(shí)成像
＊脅迫損傷的早期檢測
＊不連接顯微鏡即可測量綠色熒光蛋白（gfp）熒光
＊可測量葉片吸光系數(shù)

測量參數(shù)

＊以上所有參數(shù)均可成像
＊吸光系數(shù)abs和新參數(shù)ql、y(npq)和y(no)的成像是imaging-pam*的
＊生態(tài)毒理學(xué)研究中，選一個(gè)參考點(diǎn)，可以直接求出其它處理（如農(nóng)藥）的受抑制程度inh.

各種熒光參數(shù)的成像是將0.0（黑色）至1.0（紫色）的數(shù)值轉(zhuǎn)換成顏色來顯示的。

應(yīng)用范圍

＊環(huán)境科學(xué)
＊水生生物學(xué)
＊海洋與湖沼學(xué)
＊生態(tài)毒理學(xué)
＊園藝學(xué)
＊農(nóng)業(yè)科學(xué)
＊林學(xué)

dcmu在葉片中的滲透過程

m系列imaging-pam不同版本介紹

maxi-版
大探頭，成像面積10×13 cm

調(diào)制熒光成像系統(tǒng)的maxi-探頭利用300 w的led陣列，可以在10×13 cm的面積上提供均為的調(diào)制測量光、光化光和飽和脈沖光。該探頭的支架上配備特制護(hù)眼遮光罩，可以在保護(hù)眼睛的同時(shí)觀測到紅色熒光的變化。

walz提供兩種數(shù)碼相機(jī)ccd供選擇。用戶若需要高清晰度，推薦選擇imag-max/k[2/3" chip, 1392×1040象素， 4象素組合（binning）技術(shù)]。標(biāo)準(zhǔn)應(yīng)用可選擇imag-max/k2（1/2", 640×480象素），可與imag-max/k2z物鏡（f1.0/f=8-48mm）結(jié)合使用。


測量盆栽植物	測量離體葉片	測量微藻樣品	新增鏡頭可調(diào)放大倍數(shù)


y(npq)	ps/50	f	96個(gè)微藻樣品成像

mini-版
小探頭，成像面積24×32 mm

調(diào)制熒光成像系統(tǒng)的mini-探頭采用強(qiáng)大的luxeon led陣列，包括4組（每組3個(gè)）led，均配有長波截止濾光片。配備8個(gè)紅光（650 nm）和8個(gè)近紅外（780 nm）led，用于測量葉片吸光系數(shù)的成像。

3種版本可選：
imag-min/b：藍(lán)光，450 nm，測量葉片等；
imag-min/r：紅光，620 nm，測量藍(lán)藻；
imag-min/gfp：藍(lán)光，480 nm，測量綠色熒光蛋白（gfp）

由于mini-探頭的便攜式設(shè)計(jì)，使其特別適合野外應(yīng)用。由于mini-探頭的成像面積僅為maxi-探頭的1/16，因而前者發(fā)出的zui大光強(qiáng)更大，但耗電卻小得多。mini-探頭可以安裝在光合儀gfs-3000的葉室3010-s上，同步測量全葉片氣體交換和熒光成像，并且其光源可由gfs-3000控制，達(dá)到真正的同步測量。
mini-探頭采用1/3"數(shù)碼相機(jī)ccd（640×480象素）和f1.2/f=12mm物鏡。其設(shè)計(jì)目的為測量固定距離下的熒光成像。


與光合儀gfs-3000連用	長時(shí)間測量可裝在三角架上	fm	qn

micro-版
微探頭，成像面積3.5×4.5 mm

調(diào)制熒光成像系統(tǒng)的micro-探頭是一個(gè)極便攜的探頭，采用整合式cosmicar-pentax cctv物鏡（f1.4/f=16mm），直接安裝在數(shù)碼ccd（1/3" chip, 640×480像素）上。

micro-探頭只配備一個(gè)luxeon led（藍(lán)光，450 nm）和一個(gè)特制雙色分光鏡，類似于落射熒光顯微鏡。

盡管成像面積只有3.5×4.5 mm，但45倍的放大率卻允許對(duì)葉片熒光成像的異質(zhì)性分析達(dá)到支脈（minor veins）級(jí)。同時(shí)還可提供一個(gè)特制版本用于測量gfp的成像。

micro-探頭還可安裝在標(biāo)準(zhǔn)版imaging-pam（2001年設(shè)計(jì)）主機(jī)上。該探頭提供x-y軸可調(diào)的樣品臺(tái)。其設(shè)計(jì)目的為測量固定距離下的熒光成像。


fo	gfp成像	微探頭直接安裝在ccd上

microscopy-版
顯微探頭，成像面積130×150 um

必須與特制落射熒光顯微鏡（hund或zeiss）結(jié)合使用，該顯微鏡可以提供激發(fā)光并檢測熒光

imag-max/k（數(shù)碼相機(jī)ccd）[1392×1040象素，4象素組合（binning）技術(shù)]可以提供高靈敏度。

探頭標(biāo)準(zhǔn)配置是一個(gè)*luxeon led（450-480 nm），用于提供測量光、光化光和飽和脈沖。目前已可提供rgb探頭（紅-綠-藍(lán)-白led光源），它采用了phyto-pam技術(shù)，可以顯微鏡下自動(dòng)對(duì)藍(lán)藻、綠藻、硅/甲藻、紅藻進(jìn)行分類并測量光合作用。


分類，紅色為硅藻，綠色為絲狀綠藻	光合，fv/fm活性，可區(qū)分細(xì)胞不同部位的活性

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