Photon Imaging System Research Articles

Quantitative analysis of luciferase activity of viral and hybrid promoters in bovine preimplantation embryos.

This study was conducted to investigate quantitatively the luciferase activity of gene constructs with viral and hybrid enhancers and promoters in bovine preimplantation embryos by using firefly luciferase reporter genes. In Experiment I, to examine the stability of the luciferase, bioluminescence intensity of bovine embryos injected with the luciferase gene driven by the SV40 early promoter and enhancer (SVEluc) was measured with a luminometer at 2 days after microinjection. The results indicated that the bioluminescence could be analysed at any time within 30 min because the luciferase activity was constant during the measurement period from 5 to 30 min. In Experiment II, the luciferase expression of fertilized oocytes injected with four gene constructs (TKEluc, TK6WEluc, SVEluc, and Miwluc) was analysed by using a photon imaging system at 2 or 6 days following microinjection. The results from Experiment II indicated that the reporter gene governed by the Miw promoter (RSV LTR and chicken beta-actin promoter) was expressed more intensively in bovine morulae and blastocysts than three other gene constructs. In Experiment III, the effect of SV40 enhancer was investigated when fused downstream to the luciferase cDNA of the Miwluc vector. The results showed that SV40 enhancer further activated the luciferase activity of the Miw promoter in bovine preimplantation embryos. It was concluded, therefore, that the Miw promoter together with the SV40 enhancer would confer the strongest expression of the firefly luciferase reporter gene among the gene constructs tested in preimplantation bovine embryos.

Three-photon excitation fluorescence imaging of biological specimens using an all-solid-state laser

We demonstrate that three-photon excitation images of both fixed and living biological specimens can be readily obtained using an all-solid-state Nd:YLF laser excitation source. Optically sectioned images of fixed Caenorhabditis elegans embryos stained with DAPI and embryos triple-labeled with DAPI, fluorescein and Texas Red are presented. Time series images of a living LLC-PK cell stained with Hoechst 33342 during the progression from metaphase to telophase are also presented. The mode of excitation was inferred from the power-law of anthracene and Hoechst 33342 fluorescence versus incident laser power and an axial resolution comparison of anthracene fluorescence with two-photon excited Calcium Crimson fluorescence. Multiphoton excitation imaging is an attractive method for optically sectioning live specimens because of the lower levels of phototoxicity produced compared to other optical sectioning techniques. The combination of two- and three-photon excitation extends the capabilities of a multiple- photon imaging system since a single wavelength can provide localized excitation of a wide variety of fluorophores whose collective emission spectra can span the entire visible spectrum.

Three‐photon excitation fluorescence imaging of biological specimens using an all‐solid‐state laser

We demonstrate that three-photon excitation images of both fixed and living biological specimens can be readily obtained using an all-solid-state Nd:YLF laser excitation source. Optically sectioned images of fixed Caenorhabditis elegans embryos stained with DAPI and embryos triple-labeled with DAPI, fluorescein and Texas Red are presented. Time series images of a living LLC-PK cell stained with Hoechst 33342 during the progression from metaphase to telophase are also presented. The mode of excitation was inferred from the power-law of anthracene and Hoechst 33342 fluorescence versus incident laser power and an axial resolution comparison of anthracene fluorescence with two-photon excited Calcium Crimson fluorescence. Multiphoton excitation imaging is an attractive method for optically sectioning live specimens because of the lower levels of phototoxicity produced compared to other optical sectioning techniques. The combination of two- and three-photon excitation extends the capabilities of a multiple- photon imaging system since a single wavelength can provide localized excitation of a wide variety of fluorophores whose collective emission spectra can span the entire visible spectrum.

New approaches in medical imaging using plastic scintillating detectors

A small animal imaging camera was built in our laboratory, using-fast plastic scintillating detectors ( τ = 2–4 ns) and position sensitive photomultipliers (Hamamatsu) digitized using flash ADCs. Pinhole collimators were used for 125I imaging to achieve submillimeter resolution with scintillating plates of 28 mm radius and 1.5 mm thickness. A high resolution PET module was constructed with arrays of 1.0 mm diameter plastic scintillating fibers. The feasibility of high resolution imaging was demonstrated by the study of brain blood flow in a rat using 125I IMP in single photon detection mode and with 64Cu PTSM by using PET mode. Construction of single photon and positron emission tomographic imaging systems for small animals and subsequently for human imaging is in progress.

A Monte Carlo program for the simulation of scintillation camera characteristics

There is a need for mathematical modelling for the evaluation of important parameters for photon imaging systems. A Monte Carlo program which simulates medical imaging nuclear detectors has been developed. Different materials can be chosen for the detector, a cover and a phantom. Cylindrical, spherical, rectangular and more complex phantom and source shapes can be simulated. Photoelectric, incoherent, coherent interactions and pair production are simulated. Different detector parameters, e.g. the energy pulse-height distribution and pulse pile-up due to finite decay time of the scintillation light emission, can be calculated. An energy resolution of the system is simulated by convolving the energy imparted with an energy-dependent Gaussian function. An image matrix of the centroid of the events in the detector can be simulated. Simulation of different collimators permits studies on spatial resolution and sensitivyt. Comparisons of our results with experimental data and other published results have shown good agreement. The usefulness of the Monte Carlo code for the accurately simulation of important parameters in scintillation camera systems, stationary as well as SPECT (single-photon emission computed tomography) systems, has been demonstrated.

Low Contrast Detectability and Contrast/Detail Analysis in Medical Ultrasound

Abstruct-The first- and second-order statistics of envelope detected ultrasound (US)B-mode images for the case of a scattering phantom with many scatteiers per resolution cell have been previously derived. These characteristics are integrated over the region of a simulated focal (disk) lesion and the signal-to-noise ratio (SNR) for lesion detectability is obtained. This SNR requires the average number of independent speckle cells over the lesion area (analogous to the number of x-ray photons over the lesion area in incoherent light or x-ray imaging). This number is obtained from our autocorrelation analysis (second-order statistics). By setting the SNR expression equal to the threshold value SNRT required to detect a lesion in the presence of speckle noise, the dependence of lesion contrast on lesion diameter at threshold is found, i.e., the contrast/detail function. This is a simple inverse relation for ideal observers of US B-scans. It is also found that the contrast/detail results for envelope detectionin diagnostic ultrasound are almost identical with the results for square law detection (the usual laser case) with the latter serving as an upper limit for performance in lesion detection. Finally, the results of human observer performance using a contrast/ detail phantom are compared with the predictions for optimal or ideal 2cm Fig. 1. Schematic of contrast/detail phantom for diagnostic ultrasound. performance. The results are comparable with results for photon imaging systems, with valuesof the SNR at threshold in the neighborhood of 2-3.

The Ranicon: A resistive anode image converter

A versatile photon and charged particle imaging system is described. The Ranicon employs a microchannel electron multiplier plate to convert each detected event into a charge signal. This charge pulse is proximity focused onto a large area resistive anode plate equipped with pickup electrodes on its edges. Each event is located electronically by the ratios of the charges collected at the edges or by the differences of the signals' rise times. One or two dimensional pictures are built up by storing events digitally (e.g., a core memory) or in analog form (e.g., a storage oscilloscope). Compact laboratory models have been constructed and tested. Operating characteristics, applications, limitations, and advantages of the Ranicon are discussed.