研究目的
Investigating the spectroscopic characteristics of cancer tissues using GPU accelerated real-time confocal fluorescence lifetime imaging microscopy (FLIM) based on the analog mean-delay (AMD) method.
研究成果
The study demonstrated a real-time confocal AMD-FLIM system for spectroscopic sensing of cancer tissues, achieving a frame rate of ~13 fps for real-time confocal FLIM with 200 × 200 pixels.
研究不足
Photobleaching and intensity variation occurred in the fluorescence intensity image.
1：Experimental Design and Method Selection:
The study utilized the analog mean-delay (AMD) method for high-speed confocal fluorescence lifetime imaging.
2：Sample Selection and Data Sources:
Alexa Fluor 488 diluted by phosphate buffered saline (PBS) on a glass slide was used.
3：List of Experimental Equipment and Materials:
Diode pulse laser, dichroic mirror, optical short pass filter, x-y scanner, objective lens, photomultiplier tube, digitizer, data acquisition board, GPU.
4：Experimental Procedures and Operational Workflow:
The system synchronized the x-y scanner, pulse laser, and digitizer for data acquisition.
5：Data Analysis Methods:
The fluorescence lifetime was calculated using the AMD method.
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Dichroic mirror
MD499
Thorlabs
Reflects the laser beam towards the sample and transmits the fluorescence signal
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Optical short pass filter
FF01-498/SP-25
Semrock
Filters out wavelengths longer than 498 nm
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Photomultiplier tube
H10720-01
Hamamatsu
Detects fluorescence signals
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GPU
GeForce GTX TITAN Black
NVIDIA
Accelerates data processing
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Diode pulse laser
479 nm, 8 MHz, 30 ps, 0.8 mW
Excitation source for fluorescence imaging
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Digitizer
PCI-5114
National Instruments
Acquires electric pulse signals from the PMT
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Data acquisition board
PCIE-6353
National Instruments
Controls the digitizer and synchronizes the system
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