When using the 40X 1.0NA water dipping objective from Zeiss we find that these two image planes are offset from each other by approximately 150 microns. Changing the focal length(s) and thus the expansion ratio of the scan lens and/or tube lens can also modify the beam diameter. Anal Bioanal Chem. 1–15 (2013).9.Zhu, H., Sikora, U. & Ozcan, A. Quantum dot enabled detection of Escherichia coli using a cell-phone.
The face-value comparison is valuable though, as equating all or most of the variables between the systems nullifies the test. It was the overall design and the parts chosen for TIMAHC that was of interest to compare to out-of-the-box commercial systems to gauge performance differences. Bottom) xy motor stage and single-axis turning mirror are shown. Thanks to Craig Brideau from the laboratory of Dr. Peter Stys in helping with the signal-to-noise data collection on the Nikon A1 multi-photon microscope.
Either the sample stage and condenser height can be configured and optimized for a particular objective, or one can add extension collars (Thorlabs, part# SM1LXX) to the shorter objectives to equalize the parfocal distance. Meanwhile, the NREA algorithm effectively reduces the noise while preserving the desired signal. Nat Methods 2: 932–940 doi:10.1038/nmeth818. 2. So PT, Dong CY, Masters BR, Berland KM (2000) Two-photon excitation fluorescence microscopy. Noise intensity was calculated by spatially filtering and averaging pixel intensity outside the signal area.Full size imageThe NREA algorithm is based on suppression of the random noise signal by ensemble averaging while preserving the additive nature of the desired signal.