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M系列調(diào)制葉綠素?zé)晒獬上裣到y(tǒng)IMAGING-PAM

簡要描述:全葉片熒光成像,精確到每個像素,光合作用橫向異質(zhì)性分析,多達(dá)17個熒光參數(shù),測量熒光誘導(dǎo)、暗弛豫、光響應(yīng)曲線等,代表了調(diào)制葉綠素?zé)晒饧夹g(shù)的未來發(fā)展趨勢。

  • 更新日期:2024-11-11
  • 訪  問  量: 11324

詳細(xì)介紹


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點(diǎn)的光合活性。利用一個點(diǎn)的數(shù)據(jù)代表一個葉片,利用一個葉片代表一個植株,進(jìn)而代表一個群體(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

放大6

放大45

放大130-1300


功 能
* 一個主機(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
* *相同的條件下同時測量多個樣品(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)做多個微藻樣品的同時成像
* 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)
* 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)led,均配有長波截止濾光片。配備8個紅光(650 nm)和8個近紅外(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luxeon led(藍(lán)光,450 nm)和一個特制雙色分光鏡,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葉片熒光成像的異質(zhì)性分析達(dá)到支脈(minor veins)級。同時還可提供一個特制版本用于測量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。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*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),可以顯微鏡下自動對藍(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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50. aldea m, hamilton jg, resti jp, zangerl ar, berenbaum mr, frank td, delucia eh: comparison of photosynthetic damage from arthropod herbivory and pathogen infection in understory hardwood saplings. oecologia 2006;149:221-232.
51. bonfig kb, schreiber u, gabler a, roitsch t, berger s: infection with virulent and avirulent p. syringae strains differentially affects photosynthesis and sink metabolism in arabidopsis leaves planta 2006;225:1-12.
52. dima e, manetas y, psaras gk: chlorophyll distribution pattern in inner stem tissues: evidence from epifluorescence microscopy and reflectance measurements in 20 woody species trees 2006;20:515-521.
53. escher bi, quayle p, muller r, schreiber u, mueller jf: passive sampling of herbicides combined with effect analysis in algae using a novel high-throughput phytotoxicity assay (maxi-imaging-pam). journal of environmental monitoring 2006;8:456-464.
54. heddad m, norén h, reiser v, dunaeva m, andersson b, adamska i: differential expression and localization of early light-induced proteins in arabidopsis thaliana. plant physiology 2006:in press.
55. hölzl g, witt s, kelly aa, zähringer u, warnecke d, dörmann p, heinz e: functional differences between galactolipids and glucolipids revealed in photosynthesis of higher plants. proc. natl. acad. sci. usa 2006;103:7512-7517.
56. ivanov ag, hendrickson l, krol m, selstam e, öquist g, hurry v, huner npa: digalactosyl-diacylglycerol deficiency impairs the capacity for photosynthetic intersystem electron transport and state transitions in arabidopsis thaliana due to photosystem i acceptor-side limitations. plant cell and physiology 2006;47:1146-1157.
57. kaiser h, grams tee: rapid hydr-opassive opening and subsequent active stomatal closure follow heat-induced electrical signals in mimosa pudica. journal of experimental botany 2006;57:2087-2092.
58. kuster a, altenburger r: development and validation of a new fluorescence-based bioassay for aquatic macrophyte species. chemosphere 2006;67:194-201.
59. lohmann a, schottler ma, brehelin c, kessler f, bock r, cahoon eb, dormann p: deficiency in phylloquinone (vitamin k1) methylation affects prenyl quinone distribution, photosystem i abundance, and anthocyanin accumulation in the arabidopsis atmeng mutant. journal of biological chemistry 2006;281:40461-40472.
60. nagel ka, schurr u, walter a: dynamics of root growth stimulation in nicotiana tabacum in increasing light intensity. plant cell and environment 2006;29:1936-1945.
61. petit a-n, vaillant n, boulay m, clément c, fontaine f: alteration of photosynthesis in grapevines affected by esca. phytopathology 2006;96:1060-1066.
62. pieruschka r, schurr u, jensen m, wolff wf, jahnke s: lateral diffusion of co2 from shaded to illuminated leaf parts affects photosynthesis inside homobaric leaves. new phytologist 2006;169:779-788.
63. swarbrick pj, schulze-lefert p, scholes jd: metabolic consequences of susceptibility and resistance (race-specific and broad-spectrum) in barley leaves challenged with powdery mildew. plant cell and environment 2006;29:1061-1076.
64. vopel k, hawes i: photosynthetic performance of benthic microbial mats in lake hoare, antarctica. limnology and oceanography 2006;51:1801-1812.
65. 蔡馬, 賀立紅, 梁紅: 2種銀杏葉片葉綠素?zé)晒馓匦缘谋容^. 安徽農(nóng)業(yè)科學(xué) 2006;34:3322-3324.
66. 鞏擎柱, 呂金印, 徐柄成, 李鳳民, 張海波: 水分脅迫和種植方式對小麥葉綠素?zé)晒鈪?shù)及水分利用效率的影響. 西北農(nóng)林科技大學(xué)學(xué)報(bào) 2006;34:83-87.
67. 郭學(xué)民, 王貴禧, 高榮孚, 梁麗松: 果實(shí)表皮毛的掃描電鏡觀察及其對果實(shí)表面熒光特性的影響. 內(nèi)蒙古農(nóng)業(yè)大學(xué)學(xué)報(bào) 2006;27:43-47.
68. 賀立紅, 賀立靜, 顧群, 梁紅: 銀杏同一葉片不同部位葉綠素?zé)晒馓匦缘难芯? 北方園藝 2006:27-29.
69. 賀立紅, 賀立靜, 梁紅: 銀杏不同品種葉綠素?zé)晒鈪?shù)的比較. 華南農(nóng)業(yè)大學(xué)學(xué)報(bào) 2006;27:43-46.
70. aldea m, hamilton jg, resti jp, zangerl ar, berenbaum mr, delucia eh: indirect effects of insect herbivory on leaf gas exchange in soybean. plant cell and environment 2005;28:402-411.
71. baek m-h, kim j-h, chung by, kim j-s, lee is: alleviation of salt stress by low dose g-irradiation in rice. biologia plantarum 2005;49:273-276.
72. borisjuk l, nguyen th, neuberger t, rutten t, tschiersch h, claus b, feussner i, webb ag, jakob p, weber h, wobus u, rolletschek h: gradients of lipid storage, photosynthesis and plastid differentiation in developing soybean seeds. new phytologist 2005;163:761-776.
73. gog l, berenbaum mr, delucia eh, zangerl ar: autotoxic effects of essential oils on photosynthesis in parsley, parsnip, and rough lemon. chemoecology 2005;15:115-119.
74. kühl m, chen m, ralph pj, schreiber u, larkum awd: a niche for cyanobacteria containing chlorophyll d. nature 2005;433:820.
75. lautner s, grams tee, matyssek r, fromm j: characteristics of electrical signals in poplar and responses in photosynthesis. plant physiology 2005;138:2200-2209.
76. manetas y, pfanz h: spatial heterogeneity of light penetration through periderm and lenticels and concomitant patchy acclimation of corticular photosynthesis trees 2005;19:409-414.
77. marjanovic z, uwe n, hampp r: mycorrhiza formation enhances adaptive response of hybrid poplar to drought annals of the new york academy of sciences 2005;1048:496-499.
78. podola b, melkonian m: se-lective real-time herbicide monitoring by an array chip biosensor employing diverse microalgae. journal of applied phycology 2005;17:261-271.
79. ralph pj, macinnis-ng cmo, frankart c: fluorescence imaging application: effect of leaf age on seagrass photokinetics. aquatic botany 2005;81:69-84.
80. ralph pj, schreiber u, gademann r, kühl m, larkum awd: coral photobiology studied with a new imaging pulse amplitude modulated fluorometer. journal of phycology 2005;41:335-342.
81. berger s, papadopoulos m, schreiber u, kaiser w, roitsch t: complex regulation of gene expression, photosynthesis and sugar levels by pathogen infection in tomato. physiologia plantarum 2004;122:419-428.
82. hill r, larkum awd, frankart c, kühl m, ralph pj: loss of functional photosystem ii reaction centres in zooxanthellae of corals exposed to bleaching conditions: using fluorescence rise kinetics. photosynthesis research 2004;82:59-72.
83. hill r, schreiber u, gademann r, larkum awd, kühl m, ralph pj: spatial heterogeneity of photosynthesis and the effect of temperature-induced bleaching conditions in three species of corals. marine biology 2004;144:633-640.
84. podola b, nowack ecm, melkonian m: the use of multiple-strain algal sensor chips for the detection and identification of volatile organic compounds. biosensors and bioelectronics 2004;19:1253-1260.
85. koziolek c, grams tee, schreiber u, matyssek r, fromm j: transient knockout of photosynthesis mediated by electrical signals. new phytologist 2003;161:715-722.
86. schreiber u, walz h, kolbowski j: propagation of spatial variations of chlorophyll fluorescence parameters in dandelion leaves induced by spot laser heating. pam news 2003.

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