C13440

The mouse caudal epididymis was removed and bluntly dissociated in 1 mL high-saline solution (HS: 135 mM NaCl, 5 mM KCl, 1 mM MgSO₄, 2 mM CaCl₂, 20 mM HEPES, 5 mM glucose, 10 mM lactic acid and 1 mM Na-pyruvate, adjusted to pH 7.4 with NaOH) to release the sperm at 37 °C for 10 min. Then, the sperm suspension was transferred to petri dishes coated with 0.05% polylysine in advance, so that the sperm head was fixed, while the tail could swing freely. The flagellum oscillation of sperm was recorded at 200 fps for 3 s and multiple images were generated using a Hamamatsu digital camera C13440 (Hamamatsu, Tokyo, Japan) equipped in a Nikon microscope. Fuji software was used to synthesize a superimposed image to track the waveform of the sperm flagellum, as previously represented [48 (link)].

We assembled an imaging system in a black lead box. The x-ray source used in the system was M237 (50 kV, Newton Scientific) with Au target. The Cs₅Cu₃Cl₆I₂ film was placed on a reflector (CCM1-G01, Thorlabs), and the scintillation light was deflected to a CMOS camera (C13440, Hamamatsu) with pixel size of 6.5 μm by 6.5 μm. For the demonstration of chip pipelining inspection, a chip was fixed to a scanning stage (PSA200-11-X, Zolix) at a step rate of 4.5 mm s⁻¹. The camera was set at automatic acquisition mode. For the angiography simulation, a processing approach typical of digital subtraction angiography was applied. A “mask image” was captured before the injection of the contrast agent. This mask image was then subtracted from the subsequent image taken after contrast injection, yielding a clear angiographic image. The pixel values of scintillation photon noise and the camera’s electronic noise were calculated as 6.4 (fig. S26).
DQE was calculated with MIQuaELa software and measured following the guidance of the international standard IEC62372. The noise power spectrum (NPS; fig. S27) and MTF were measured with RQA3 beam, which was produced form a tube (Leo, Varex Imaging) voltage of 50 kV. The detector responses were measured under several different doses to obtain signal transfer property. The absorption dose of 2.5 μGy_air was used to measure the NPS of the detector.

We assembled a homemade imaging system in a black lead box. The x-ray source used 435 in the system was M237 (50 kV, Newton Scientific) with Au target. The BA₁₀EuI₁₂/PS film was placed on a reflector (CCM1-G01, Thorlabs), and the scintillation light was deflected to a 437 CMOS camera (C13440, Hamamatsu) with a pixel size of 6.5 × 6.5 μm².

To conduct imaging experiments, AuNPs were used as a representative sample. In total, 30 μL of the AuNPs solution was applied onto the microlens chip, followed by an observation with an inverted optical microscope equipped with a white light source (white LED light source, ZEISS Colibri, ZEISS, Oberkochen, Germany), as depicted in Figure 1. A 40× objective (ZEISS LD A-Plan) with a numerical aperture (NA) of 0.55 was used. The objective was focused on the imaging plane of the microlens. The backscattering signal of AuNPs was collected by the objective and finally transmitted to a sCMOS camera (Hamamatsu C13440, Hamamatsu, Shizuoka, Japan). The exposure time was set between 166 μs and 20 ms, and the region of interest (ROI) was adjusted accordingly. The intensity of the AuNP is represented by the grayscale values measured from the images. Only the AuNPs located in the ROI were included in the statistics.

For immunostaining, 3D images were taken using an Olympus IX83 microscope and Hamamatsu C13440 digital camera. All images in the same experiments were captured with the same exposure time. The foci were selected by Imaris (Oxford Instruments) or cellSens (Olympus) software. Colocalization events were counted using the Imaris spot detection function in conjunction with the colocalization channel. For the quantification of telomere intensity, 3D images were projected to 2D with maximum intensity projection function, and the telomere foci were selected using the Imaris surface function. For quantification of TERRA intensity, TERRA signals were selected using cellSens. To calculate the intensity of XPF foci at telomeres, the signals of TRF2 foci were selected as ROI, the intensity of XPF was counted on the ROI of TRF2. Statistical significance determined by student’s t-test or Mann-Whitney test and other parameters were analyzed by GraphPad Prism.