Fluorescence lifetime imaging can reveal the high-resolution structure of various biophysical and chemical parameters in a microenvironment quantitatively.However,the depth of imaging is generally limited to hundreds ...Fluorescence lifetime imaging can reveal the high-resolution structure of various biophysical and chemical parameters in a microenvironment quantitatively.However,the depth of imaging is generally limited to hundreds of micrometers due to aberration and light scattering in biological tissues.This paper introduces an iterative multi-photon adaptive compensation technique(IMPACT)into a two-photon fluorescence lifetime microscopy system to successfully overcome aberrations and multiple scattering problems in deep tissues.It shows that 400 correction modes can be achieved within 5 min,which was mainly limited by the frame rate of a spatial light modulator.This system was used for high-resolution imaging of mice brain tissue and live zebrafish,further verifying its superior performance in imaging quality and photon accumulation speed.展开更多
基金supported by the National Key Research and Development Program of China(No.2021YFF0502900)the National Natural Science Foundation of China(Nos.62175163,62225505,61935012,61835009,62127819,and 62205220)+2 种基金the Shenzhen Key Projects(No.JCYJ20200109105404067)the Shenzhen Talent Innovation Project(No.RCJC20210706091949022)the Shenzhen Science and Technology Planning Project(No.ZDSYS20210623092006020)。
文摘Fluorescence lifetime imaging can reveal the high-resolution structure of various biophysical and chemical parameters in a microenvironment quantitatively.However,the depth of imaging is generally limited to hundreds of micrometers due to aberration and light scattering in biological tissues.This paper introduces an iterative multi-photon adaptive compensation technique(IMPACT)into a two-photon fluorescence lifetime microscopy system to successfully overcome aberrations and multiple scattering problems in deep tissues.It shows that 400 correction modes can be achieved within 5 min,which was mainly limited by the frame rate of a spatial light modulator.This system was used for high-resolution imaging of mice brain tissue and live zebrafish,further verifying its superior performance in imaging quality and photon accumulation speed.
基金The project of Hebei Administration of Traditional Chinese Medicine (Grant No. 2020167)the project of the Second Hospital of Hebei Medical University (Grant No. 2HS202014)the Chinese Medicine Association (Grant No. CMEI2019KPYJ00142)
