浮游植物熒光儀Phyto-PAM
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浮游植物熒光儀Phyto-PAM款可自動(dòng)對(duì)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而獲得首屆光合作用協(xié)會(huì)(ispr)創(chuàng)新獎(jiǎng)
1983年，walz公司*科學(xué)家、德國(guó)烏茲堡大學(xué)的ulrich schreiber教授設(shè)計(jì)制造了**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(cè)器，因此只能檢測(cè)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(zhǎng)調(diào)制熒光儀的構(gòu)想。1988年，schreiber教授和他的博士后kolbowski博士*次設(shè)計(jì)出了16波長(zhǎng)(led)熒光分光光度計(jì)。
1995年，schreiber教授和kolbowski博士一起設(shè)計(jì)出了**臺(tái)可對(duì)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(zhǎng)的led作為光源，利用光電倍增管作為檢測(cè)器，可以對(duì)水樣中的藍(lán)藻、綠藻、硅藻/甲藻自動(dòng)分類，并分別測(cè)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(cè)器，因此檢測(cè)限達(dá)到 0.1
μg l-1 chl。根據(jù)研究對(duì)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(cè)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(cè)量原理是基于對(duì)調(diào)制測(cè)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(cè)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(cè)量光脈沖在高頻率下交替應(yīng)用，就可以獲得4種波長(zhǎng)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(cè)量它們的光合活性和葉綠素含量。
phyto-pam可以對(duì)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(duì)它們進(jìn)行區(qū)分。要想對(duì)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(cè)量熒光。目前walz公司正結(jié)合“泵”和“探針”法開(kāi)發(fā)對(duì)微藻分類更多、更精確的儀器。phyto-pam還可以測(cè)量這些藻類的葉綠素濃度(檢測(cè)限為0.1
μg l-1 chl)。phyto-pam更加強(qiáng)大的功能是可以探測(cè)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(cè)器，可以檢測(cè)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) 可對(duì)浮游植物自動(dòng)分類的調(diào)制葉綠素?zé)晒鈨x
2) 4波長(zhǎng)光源：470、520、645和665 nm
3) 對(duì)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(cè)附著藻類/大型藻類的系統(tǒng)(iii)
5) 靈敏度高，檢測(cè)限為0.1μg 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(cè)量后根據(jù)水體藻類組成利用優(yōu)勢(shì)種(一種或多種)的參考光譜校對(duì)實(shí)驗(yàn)結(jié)果
功能：
1) 可對(duì)藍(lán)藻、綠藻和硅/甲藻自動(dòng)分類(定性)
2) 可自動(dòng)測(cè)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(cè)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(cè)量光合作用的量子產(chǎn)量和相對(duì)電子傳遞速率
5) 可自動(dòng)記錄量子產(chǎn)量和相對(duì)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：
浮游植物熒光儀Phyto-PAM 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(cè)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(duì)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(cè)器可以組成3套不同的測(cè)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(cè)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(kāi)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(chǎng)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(cè)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(cè)單元phyto-ed中。phyto-ed密封防水。系統(tǒng)ii在野外或在船上工作，當(dāng)然室內(nèi)也*可以使用。
系統(tǒng)iii
光纖型版本，利用phyto-edf檢測(cè)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(cè)單元phyto-edf可以檢測(cè)所有生長(zhǎng)在表面的光合生物的光合作用。比較適合的測(cè)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(cè)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*可滿足實(shí)驗(yàn)要求。
| ● 基礎(chǔ)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(cè)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(cè)單元phyto-ed | ● | ||
| 光纖型激發(fā)-檢測(cè)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 | ○ |
浮游植物熒光儀Phyto-PAM 技術(shù)參數(shù)：
測(cè)量光：波長(zhǎng)470、520、645和665 nm的測(cè)量光led。
光化光：波長(zhǎng)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(zhǎng)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(cè)：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(cè)量參數(shù)：ft, f(或fo), fm(或 fm’), δf, y(δf/ fm’或fv/fm), etr和chl濃度等。
環(huán)境溫度：-5～+45 ℃，已在極地成功應(yīng)用。
部分文獻(xiàn)
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