pco.flim 荧光寿命分析系统

发布时间:2021-04-20 01:54:59

pco.flim相机系统是采用Two-Tap CMOS成像芯片的荧光寿命成像相机。相机的像素和激发光间的同步使100皮秒至100微秒间的荧光衰减时间的分析成为可能。在1008 x 1008像素分辨率上,pco.flim最多可以读取45个双帧/秒,且可支持5 kHz - 40 MHz频率范围的调制。其C-mount接口可以使改相机系统轻松地连接到任何显微镜或镜头。此外,其USB 3.0接口可让您

pco.flim相机系统是采用Two-Tap CMOS成像芯片的荧光寿命成像相机。相机的像素和激发光间的同步使100皮秒至100微秒间的荧光衰减时间的分析成为可能。在1008 x 1008像素分辨率上,pco.flim最多可以读取45个双帧/秒,且可支持5 kHz  -  40 MHz频率范围的调制。

其C-mount接口可以使改相机系统轻松地连接到任何显微镜或镜头。此外,其USB 3.0接口可让您将相机连接到各种计算机。这会为您的实验和研究节省了大量的精力和成本。

pco.flim激光器是一种均匀照明的光源,设计用于pco.flim。它的数字调制频率范围为0  -  250 MHz。您也可以订购pco.flim激光用于宽场荧光显微镜照明,或者用于全内反射荧光(TIRF)显微镜,光片荧光显微镜(LSFM)和共焦旋转盘显微镜。

您可在多种不同波长和输出功率的激光。如果您样本的发光寿命在几纳秒的范围内,则选择激光作为光源使至关重要的。如果您使用的是尼康显微镜,我们的激光也提供相匹配的尼康双安全快门选项。元奥仪器是德国PCO.AG公司在中国区的指定授权代理商,负责德国PCO.AG产品在中国市场的推广、销售、技术支持等工作,为客户提供性价比高的产品。 欢迎您来电咨询!


  • 100 ps到100µs的寿命测量

  • 调制频率从5千赫兹到40兆赫

  • 500千赫兹到40兆赫外部调制信号

  • 正弦/矩形的调制信号波形

  • 像素分辨率1008 x 1008

  • 频域FLIM

  • USB 3.0接口

  • 量子效率39%

  • 动态范围1000:1

  • 读出噪音45 e- rms

  • 90帧每秒 (2 tap读出)

  • 可选曝光时间为10ns到10秒

  • 无振动水冷

  • 特殊的测量与分析软件


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

  • selectable 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 selectable exposure times

  • vibration-free water cooling

  • special software for measurement and analysis

注:产品信息若有变更恕不另行通知  / 德国PCO公司中国指定代理商---元奥仪器






分辨率 ( x )


1008 x 1008



5.6 x 5.6


5 kHz - 40 MHz


500 kHz - 40 MHz




动态范围 A/D




pco.flim 荧光寿命成像系统装配图:


注:产品信息若有变更恕不另行通知  / 德国PCO公司中国指定代理商---元奥仪器


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 Göttingen).


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

Seemlesly integrated into 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 Göttingen).

注:产品信息若有变更恕不另行通知  / 德国PCO公司中国指定代理商---元奥仪器


1. Frequency Domain FLIM with pco.flim Modulated CMOS Camera for Fluorescence Lifetime Microscopy

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.



Hongtao Chen, Gerhard Holst, and Enrico Gratton
Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering University of California, Irvine, California Science & Research, PCO AG, Kelheim, Germany

Website: Researchgate

Related Files:

2. LLIM of Chemical Sensors with pco.flim

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 Moßhammer, Vincent V. Scholz, Sergey M. Borisov, Gerhard Holst and Michael Kühl, Analytical Chemistry 2019

Related Files:

Luminescence Lifetime Imaging of Chemical Sensors

3. Widefield FLIM of Protoporphyrin IX with pco.flim

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.



M.T. Erkkilä, 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.
Related Files: