pco.flim 熒光壽命成像相機(jī)作為第一款采用每像素兩個(gè)讀出抽頭（Two-Tap）CMOS傳感器的頻域(FD) FLIM相機(jī)，我們的pco.flim提供像素和激發(fā)光的同步調(diào)制，從而能夠分析亞納秒到微秒范圍內(nèi)的發(fā)光衰減時(shí)間。憑借1008 x 1008的像素分辨率，pco.flim最大可讀出45雙幀/秒，并可用于5 kHz - 40 MHz的調(diào)制頻率范圍。這款相機(jī)兼具靈活耦合和簡(jiǎn)便設(shè)置，為廣角FD-FLIM儀器的理想選擇。pco.flim相機(jī)系統(tǒng)可通過C接口靈活耦合至任何顯微鏡或鏡頭，并設(shè)有一個(gè)USB 3.0接口，可輕松將相機(jī)連接至各種計(jì)算機(jī)。pco.flim激光是一種均勻照明的光源，設(shè)計(jì)用于pco.flim。其數(shù)字調(diào)制頻率范圍為0 - 250 MHz。pco.flim激光適用于寬場(chǎng)落射熒光顯微鏡照明、全內(nèi)反射熒光（TIRF）顯微鏡、光片熒光顯微鏡（LSFM）和共聚焦轉(zhuǎn)盤顯微鏡。各種不同的波長(zhǎng)和光輸出功率任您選擇。pco.flim激光非常適合用于亞納秒到微秒范圍內(nèi)的相關(guān)發(fā)光壽命樣本。對(duì)于使用尼康微鏡的用戶，另提供尼康雙安全快門選擇。pco.flim相機(jī)系統(tǒng)是采用Two-Tap CMOS成像芯片的熒光壽命成像相機(jī)。相機(jī)的像素和激發(fā)光間的同步使100皮秒至100微秒間的熒光衰減時(shí)間的分析成為可能。其多種觸發(fā)選項(xiàng)使相機(jī)可以方便的集成于多種實(shí)際應(yīng)用。

德國(guó)PCO公司首次發(fā)布了這款采用Two-Tap CMOS技術(shù)的熒光壽命成像系統(tǒng)Flim，應(yīng)用范圍涵蓋分子特性分析和測(cè)試，F(xiàn)RET，離子濃度/PH值測(cè)量，生物組織學(xué)/病理學(xué)，壓感材料等，可廣泛應(yīng)用于生命科學(xué)、物理科學(xué)，材料科學(xué)領(lǐng)域。

主要特點(diǎn)：

• 100 ps到100μs的壽命測(cè)量

• 調(diào)制頻率從5千赫茲到40兆赫

• 500千赫茲到40兆赫外部調(diào)制信號(hào)

• 正弦/矩形的調(diào)制信號(hào)波形

• 像素分辨率1008 x 1008

• 頻域FLIM

• USB 3.0接口

• 量子效率39%

• 動(dòng)態(tài)范圍1000:1

• 讀出噪音48 e- rms

• 90幀每秒 (2 tap讀出)

• 可選曝光時(shí)間為10ns到10秒

• 無振動(dòng)水冷

• 特殊的測(cè)量與分析軟件

技術(shù)參數(shù)：

應(yīng)用案例：

Figure 1: The left photo shows the fluorescence intensity of HEK-293 cells which expressed a CFP/DJ-1 protein as control of a FRET experiment. The middle image shows the phase angle derived distribution of fluorescence lifetimes in the range of 0 – 4 ns (imageJ LUT 16 colors, colorbar 0 – 4 ns) which has been masked by an intensity filter. The range of lifetimes around 2 ns was found in all of the 26 cells, which have been measured and showed about 10% FRET efficiency compared to the pure CFP expression. The right image shows the lifetime distribution image weighted by the fluorescence intensity image (the color images are false color coded using the same colorbar and LUT) without mask (courtesy of Prof. Dr. F.S. Wouters and Dr. G. Bunt, University Medicine G?ttingen).

Figure 5:The left image shows the fluorescence intensity of a sliced daisy sample (20x air objective). The middle image shows the phase angle derived distribution of fluorescence lifetimes in the range of 0 - 4 ns (imageJ LUT 16 colors, colorbar 0-4 ns). The difference in fluorescence lifetimes clearly allows to differentiate the cells and the pollen grains. The right image shows the lifetime distribution image weighted by the fluorescence intensity image (the color images are false color coded, the modulation frequency was 30 MHz) (courtesy of Prof. Dr. F.S. Wouters and Dr. G. Bunt, University Medicine G?ttingen).

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