pco.flim相機系統是采用Two-Tap CMOS成像芯片的熒光壽命成像相機。相機的像素和激發光間的同步使100皮秒至100微秒間的熒光衰減時間的分析成為可能。在1008 x 1008像素分辨率上，pco.flim最多可以讀取45個雙幀/秒，且可支持5 kHz - 40 MHz頻率范圍的調制。
其C-mount接口可以使改相機系統輕松地連接到任何顯微鏡或鏡頭。此外，其USB 3.0接口可讓您將相機連接到各種計算機。這會為您的實驗和研究節省了大量的精力和成本。
pco.flim激光器是一種均勻照明的光源，設計用于pco.flim。它的數字調制頻率范圍為0 - 250 MHz。您也可以訂購pco.flim激光用于寬場熒光顯微鏡照明，或者用于全內反射熒光（TIRF）顯微鏡，光片熒光顯微鏡（LSFM）和共焦旋轉盤顯微鏡。
您可在多種不同波長和輸出功率的激光。如果您樣本的發光壽命在幾納秒的范圍內，則選擇激光作為光源使至關重要的。如果您使用的是尼康顯微鏡，我們的激光也提供相匹配的尼康雙安全快門選項。元奧儀器是德國Excelitas PCO公司高速攝像機中國總代理商，負責德國PCO.AG產品在中國市場的推廣、銷售、技術支持等工作，為客戶提供性價比高的產品。 歡迎您來電咨詢！
主要特點：
100 ps到100μs的壽命測量
調制頻率從5千赫茲到40兆赫
500千赫茲到40兆赫外部調制信號
正弦/矩形的調制信號波形
像素分辨率1008 x 1008
頻域FLIM
USB 3.0接口
量子效率39%
動態范圍1000:1
讀出噪音45 e- rms
90幀每秒 (2 tap讀出)
可選曝光時間為10ns到10秒
無振動水冷
特殊的測量與分析軟件
pco.flim
luminescence lifetime imaging camera
The pco.flim is the first Frequency Domain FLIM camera using a two tap CMOS sensor. Synchronized modulation of pixels and stimulated light enables you the analysis of luminescence decay times in the range of 100 ps – 100 μs.
With its 1008 x 1008 pixels resolution the pco.flim reads out 45 double frames/s at a max. You can use it for a modulation frequency range of 5 kHz – 40 MHz.
Using C-mount as standard the system is easy to connect to any microscope or lens. Further, the USB 3.0 interface lets you connect the camera to all kinds of computers. This saves you significant efforts and costs for operation and research.
The pco.flim laser is a homogeneously illuminating light source designed for use with the pco.flim. It features a digital modulation frequency range of 0 – 250 MHz. You can either order the pco.flim laser for widefield epifluorescence microscope illumination, or for Total-Internal-Reflection-Fluorescence (TIRF) microscopy, light sheet fluorescence microscopy (LSFM) and confocal spinning disk microscopy.
Choose between a wide range of different wavelengths and light output powers. The laser is ideally suited if your relevant luminescence lifetimes are in the range of a few nanoseconds. In case you are using a Nikon microscope, the Nikon double-safety-shutter option is also available
pco.flim laser as light source for epifluorescence measurements
selctable laser wavelengths and light output powers
100 ps – 100 μs lifetimes measurable
5 kHz – 40 MHz modulation frequencies
500 kHz – 40 MHz external modulation signals
modulation signal shape sinusoidal / rectangular
1008 x 1008 pixel resolution
frequency domain FLIM
USB 3.0 interface
39 % quantum efficiency
1000:1 dynamic range
45 double frames/s (2 tap readout)
1 ms to 2 s selctable exposure times
vibration-free water cooling
special software for measurement and analysis
注：產品信息若有變更恕不另行通知 / 德國Excelitas PCO公司高速攝像機中國總代理商—元奧儀器
技術參數：
參數 | 單位 | 設定點 | pco.flim |
分辨率 (橫 x 縱) | 像素 | 1008 x 1008 | |
像素尺寸 | μm | 5.6 x 5.6 | |
調制頻率(輸出) | 5 kHz - 40 MHz | ||
調制頻率(輸入) | 500 kHz - 40 MHz | ||
最高量子效率 | % | 39 | |
動態范圍 A/D | 位 | 14 |
芯片Qe圖：
pco.flim 熒光壽命成像系統裝配圖：
注：產品信息若有變更恕不另行通知 / 德國Excelitas PCO公司高速攝像機中國總代理商—元奧儀器
應用案例：
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| 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). | ||
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Figure 5: A Thermofisher FluoCells? prepared slide, which contains a section of mouse kidney stained with a combination of fluorescent dyes. The visible marker is Alexa Fluor? 488 wheat germ agglutinin, a green-fluorescent lectin, which was used to label elements of the glomeruli and convoluted tubules, which were excited with 488 nm at a modulation frequency of 30 MHz. The left image shows the fluorescence intensity of the mouse kidney sample (20x air objective). The middle image shows the phase angle derived distribution of fluorescence lifetimes of the Alexa Fluor 488? in the range of 1.5 – 3.5 ns (NIS Elements, colorbar 1.5 – 3.5 ns). The right image shows the lifetime distribution image weighted by the fluorescence intensity image | ||
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| Seemlesly integrated in o Nikon NIS Elements AR software to use the pco.flim camera for homodyne frequency domain fluorescence lifetime imaging with all referencing and phasor plot feedback. | Endogenous fluorescence of a Convallaria (lily of the valley) slice sample. The image shows the endogenous fluorescence lifetime distribution derived from the measured phase angle in false color coding and weighted by the fluorescence intensity. The displayed lifetimes range from 0.5 - 4 ns. | HEK-293 cells co-expressing a fusion protein with Cyan Fluorescent Protein (CFP) and with Yellow Fluorescent Protein (YFP). Dimerization of this protein is detected by FRET as judged by the reduction in CFP lifetime. The image shows the fluorescence lifetime distribution derived from the measured phase angle in false color coding and weighted by the fluorescence intensity. The displayed range is from 0 – 4 ns (see color bar, courtesy of Prof. Dr. F.S. Wouters and Dr. G. Bunt, University Medicine G?ttingen). |
注：產品信息若有變更恕不另行通知 / 德國Excelitas PCO公司高速攝像機中國總代理商—元奧儀器
應用文獻：
1. Frequency Domain FLIM with pco.flim Modulated CMOS Camera for Fluorescence Lifetime Microscopy
Abstract
Widefield frequency-domain fluorescence lifetime imaging microscopy (FDFLIM) is a fast and accurate method to measure the fluorescence lifetime of entire images. However, the complexity and high costs involved in construction of such a system limit the extensive use of this technique. PCO AG recently released the first luminescence lifetime imaging camera based on a high frequency modulated CMOS image sensor, QMFLIM2.
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Hongtao Chen, Gerhard Holst, and Enrico Gratton Website: Researchgate | |
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2. LLIM of Chemical Sensors with pco.flim
Abstract
Luminescence lifetime based imaging is still the most reliable method for generating chemical images using chemical sensor technology. However, only few commercial systems are available that enable imaging lifetimes within the relevant nanosecond to microsecond range. In this technical note we compare the performance of an older time-domain (TD) based camera system with a frequency-domain (FD) based camera system regarding their measuring characteristics and applicability for O2 and pH imaging in environmental samples and with different indicator dye systems emitting in the visible and near-infrared part of the spectrum. We conclude that the newly introduced FD imaging system delivers comparable if not better results than its predecessor, now enabling robust and simple chemical imaging based on FD luminescence lifetime measurements.
| Klaus Koren, Maria Mo?hammer, Vincent V. Scholz, Sergey M. Borisov, Gerhard Holst and Michael Kühl, Analytical Chemistry 2019 Related Files: |
3. Widefield FLIM of Protoporphyrin IX with pco.flim
Abstract
Achieving a maximal safe extent of resection during brain tumor surgery is the goal for improved patient prognosis. Fluorescence-guided neurosurgery using 5-aminolevulinic acid (5-ALA) induced Protoporphyrin IX has thereby become a valuable tool enabling a high frequency of complete resections and a prolonged progression-free survival in glioblastoma patients. We present a widefield fluorescence lifetime imaging device with 250 mm working distance working under similar conditions like surgical microscopes based on a time-of-flight based dual tap CMOS camera.
In contrast to intensity-based fluorescence imaging our method is invariant to light scattering and absorption while being sensitive to the molecular composition of the tissue.We evaluate the feasibility of lifetime imaging of Protoporphyrin IX using our system to analyze brain tumor phantoms and fresh 5-ALA labeled human tissue samples. The results demonstrate the potential of our lifetime sensing device to go beyond the limitation of current intensity-based fluorescence-guided neurosurgery.
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| M.T. Erkkil?, B. Bauer, N. Hecker-Denschlag, et al. (2018), Widefield fluorescence lifetime imaging of protoporphyrin IX for fluorescence-guided neurosurgery: an ex vivo feasibility study, J. Biophotonics, 2018;00:0–0. | |
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