phyto‑pam 全球第一款可自動(dòng)對浮游植物分類的熒光儀
schreiber教授因發(fā)明pam系列調(diào)制葉綠素?zé)晒鈨x而獲得首屆國際光合作用協(xié)會(huì)（ispr）創(chuàng)新獎(jiǎng)
1983年，walz公司首席科學(xué)家、德國烏茲堡大學(xué)的ulrich schreiber教授設(shè)計(jì)制造了全世界第一臺(tái)調(diào)制熒光儀——pam-101/102/103，并在植物生理、生態(tài)、農(nóng)學(xué)、林學(xué)、水生生物學(xué)等領(lǐng)域得到廣泛應(yīng)用，出版了大量高水平研究文獻(xiàn)。但該儀器由于采用光電二極管為檢測器，因此只能檢測高等植物、室內(nèi)培養(yǎng)的微藻等葉綠素含量較高（> 10 mg l-1）的樣品。
在pam-101/102/103出現(xiàn)的同時(shí)，schreiber教授就有了設(shè)計(jì)一臺(tái)多波長調(diào)制熒光儀的構(gòu)想。1988年，schreiber教授和他的博士后kolbowski博士第一次設(shè)計(jì)出了16波長（led）熒光分光光度計(jì)。
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1995年，schreiber教授和kolbowski博士一起設(shè)計(jì)出了全世界第一臺(tái)可對浮游植物自動(dòng)分類的調(diào)制葉綠素?zé)晒鈨xphyto-pam。phyto-pam采用調(diào)制技術(shù)，利用4種不要波長的led作為光源，利用光電倍增管作為檢測器，可以對水樣中的藍(lán)藻、綠藻、硅藻/甲藻自動(dòng)分類，并分別測量它們的葉綠素含量和光合活性。
phyto-pam由于采用光電倍增管作為檢測器，因此檢測限達(dá)到 0.1 μg l-1 chl。根據(jù)研究對象和研究目的不同，可有3套系統(tǒng)供您選擇。
phyto-pam是水域生態(tài)學(xué)、海洋與湖沼學(xué)、水質(zhì)監(jiān)測等領(lǐng)域的有效工具。
系統(tǒng)描述：
脈沖-振幅-調(diào)制（pulse-amplitude-modulation, pam）技術(shù)的測量原理是基于對調(diào)制測量光激發(fā)的熒光信號(hào)的選擇性放大。在phyto-pam浮游植物熒光儀中，微秒級(jí)的測量光脈沖是由4種不同顏色的發(fā)光二極管（led）陣列發(fā)出的：藍(lán)色（470 nm）、綠色（520 nm）、淺紅色（645 nm）和深紅色（665 nm）。不同顏色的測量光脈沖在高頻率下交替應(yīng)用，就可以獲得4種波長的光激發(fā)出的半同步的熒光信號(hào)。結(jié)合不同藻類的參考光譜（reference spectrum）就可區(qū)分不同藻類，并分別測量它們的光合活性和葉綠素含量。
phyto-pam可以對藍(lán)藻、綠藻和硅/甲藻進(jìn)行分類。由于硅藻和甲藻的色素組成差別不大，目前技術(shù)上還很難對它們進(jìn)行區(qū)分。要想對它們區(qū)分，除了考慮色素組成外，更重要的是考慮捕光色素-蛋白復(fù)合體的結(jié)構(gòu)特別是橫向截面積，這必須結(jié)合“泵”和“探針”法測量熒光。目前walz公司正結(jié)合“泵”和“探針”法開發(fā)對微藻分類更多、更精確的儀器。phyto-pam還可以測量這些藻類的葉綠素濃度（檢測限為0.1 μg l-1 chl）。phyto-pam更加強(qiáng)大的功能是可以探測自然水樣中藍(lán)藻、綠藻和硅/甲藻的光合活性和光適應(yīng)狀態(tài)。
phyto-pam采用微型光電倍增管作為檢測器，可以檢測及其微弱的靈敏變化，同時(shí)還具備強(qiáng)光自動(dòng)關(guān)閉的保護(hù)功能，因此儀器操作和維護(hù)更加容易。
特點(diǎn)：
1) 全世界第一臺(tái)可對浮游植物自動(dòng)分類的調(diào)制葉綠素?zé)晒鈨x
2) 4波長光源：470、520、645和665 nm
3) 對藍(lán)藻、綠藻和硅/甲藻進(jìn)行分類
4) 可選配室內(nèi)系統(tǒng)（i）、野外系統(tǒng)（ii）和測附著藻類/大型藻類的系統(tǒng)（iii）
5) 靈敏度高，檢測限為0.1 μg l-1 chl
6) 專業(yè)phytowin操作軟件，數(shù)據(jù)收集、分析和存貯功能強(qiáng)大
7) 用戶可利用培養(yǎng)的微藻做參考光譜，非“黑匣子”
8) 可在野外測量后根據(jù)水體藻類組成利用優(yōu)勢種（一種或多種）的參考光譜校對實(shí)驗(yàn)結(jié)果
功能：
1） 可對藍(lán)藻、綠藻和硅/甲藻自動(dòng)分類（定性）
2） 可自動(dòng)測量水樣中藍(lán)藻、綠藻和硅/甲藻的葉綠素含量（定量）和總?cè)~綠素含量
3） 可同時(shí)測量水樣中藍(lán)藻、綠藻和硅/甲藻的光合作用和總光合活性
4） 可測量光合作用的量子產(chǎn)量和相對電子傳遞速率
5） 可自動(dòng)記錄量子產(chǎn)量和相對電子傳遞速率的快速光響應(yīng)曲線
6） 用戶可做自己的參考光譜
7） 可連接記錄儀或示波器記錄原始熒光誘導(dǎo)動(dòng)力學(xué)曲線
應(yīng)用領(lǐng)域：
多用于水生生物學(xué)、水域生態(tài)學(xué)、海洋學(xué)、湖沼學(xué)、水質(zhì)監(jiān)測和預(yù)警、微藻生理學(xué)、微藻抗逆性、環(huán)境科學(xué)、生態(tài)毒理學(xué)、極地藻類（冰藻）研究等領(lǐng)域，對于了解自然水體中藻類種群的動(dòng)態(tài)變化、水華/赤潮預(yù)警、野外水體中光合作用的時(shí)空變化、校正初級(jí)生產(chǎn)力的計(jì)算等有較大幫助。
系統(tǒng)組成：
phyto-pam的主機(jī)連接不同的檢測器可以組成3套不同的測量系統(tǒng)：
系統(tǒng)i：實(shí)驗(yàn)室版本，利用光學(xué)單元ed-101us/mp和標(biāo)準(zhǔn)10×10 mm樣品杯檢測熒光
系統(tǒng)i的所有光電元件均需安裝在鐵架臺(tái)上，適合實(shí)驗(yàn)室用。但由于主機(jī)phyto-c內(nèi)置大容量電池，因此它也可以在野外或在船上使用。系統(tǒng)i的一個(gè)突出優(yōu)點(diǎn)是光學(xué)單元ed-101us/mp的開放式設(shè)計(jì)，它允許安裝不同的濾光片或不同顏色的光化光led陣列。與系統(tǒng)ii的phto-ed相比，10×10 mm樣品杯中的光場分布更加均勻。同時(shí)，系統(tǒng)i還可以連接溫度控制器us-t和微型磁力攪拌器phyto-ms。這些特點(diǎn)決定了系統(tǒng)i更加適合浮游植物光合作用的基礎(chǔ)研究。
系統(tǒng)ii：野外便攜式版本，利用phyto-ed和直徑15 mm的樣品杯檢測熒光
在系統(tǒng)ii中，所有光電元件都整合在便攜式的激發(fā)-檢測單元phyto-ed中。phyto-ed密封防水。系統(tǒng)ii最適合在野外或在船上工作，當(dāng)然室內(nèi)也完全可以使用。
系統(tǒng)iii：光纖型版本，利用phyto-edf檢測附著藻類或大型藻類的熒光
系統(tǒng)iii的光纖型激發(fā)-檢測單元phyto-edf可以檢測所有生長在表面的光合生物的光合作用。比較適合的測試材料包括附著藻類、底棲藻類、藻墊（microbial mats）和大型藻類等。由于采用光纖傳導(dǎo)信號(hào)而且測量面積小，因此靈敏度比系統(tǒng)i和ii要低。但是由于附著藻類等材料的葉綠素含量遠(yuǎn)遠(yuǎn)高于水體中的浮游植物，因此系統(tǒng)iii的靈敏度完全可滿足實(shí)驗(yàn)要求。
● 基礎(chǔ)配置
○ 可選配置 | 系統(tǒng)i
（實(shí)驗(yàn)室版）
 | 系統(tǒng)ii
（野外版）
 | 系統(tǒng)iii
（光纖版）
 |
| 主機(jī)phyto-c | ● | ● | ● |
| 測量光led陣列phyto-ml | ● | | |
| 光化光led陣列phyto-al | ● | | |
| 光電倍增管pm-101p | ● | | |
| 光學(xué)單元ed-101us/mp | ● | | |
| 工作臺(tái)st-101 | ● | | |
| 激發(fā)-檢測單元phyto-ed | | ● | |
| 光纖型激發(fā)-檢測單元phyto-edf | | | ● |
| 微型磁力攪拌器phyto-ms | ○ | | |
| 球狀微型光量子探頭us-sqs | ○ | ○ | ○ |
| 溫度控制器us-t | ○ | | |
| 攪拌器water-s | | ○ | |
技術(shù)參數(shù)：
測量光：波長470、520、645和665 nm的測量光led。
光化光：波長655 nm的led；光化光強(qiáng)度0～2000 μmol m-2 s-1 par（系統(tǒng)i和ii）或0～1300 μmol m-2 s-1 par（系統(tǒng)iii）。
飽和脈沖：波長655 nm的led；飽和脈沖強(qiáng)度4000 μmol m-2 s-1 par（系統(tǒng)i和ii）或2600 μmol m-2 s-1 par（系統(tǒng)iii）。
信號(hào)檢測：光電倍增管，帶短波截止濾光片（λ>710 nm）；選擇性鎖相放大器。
測量參數(shù)：ft, f(或fo), fm(或 fm’), δf, y(δf/ fm’或fv/fm), etr和chl濃度等。
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