High-throughput microscopy allows to high-content quantitative analysis of multiple events in a large population of cells, which is of crucial importance for many applications, such as personalized genomics, cancer diagnostics, and drug development 1, 2. Our work reveals an important capability of FPM towards high-speed high-throughput imaging of in vitro live cells, achieving video-rate QPI performance across a wide range of scales, both spatial and temporal. By using only 4 low-resolution images corresponding to 4 tilted illuminations matching a 10×, 0.4 NA objective, we present the high-speed imaging results of in vitro Hela cells mitosis and apoptosis at a frame rate of 25 Hz with a full-pitch resolution of 655 nm at a wavelength of 525 nm (effective NA = 0.8) across a wide field-of-view (FOV) of 1.77 mm 2, corresponding to a space–bandwidth–time product of 411 megapixels per second. The optical-transfer-function (OTF) analysis of FPM reveals that the low-frequency phase information can only be correctly recovered if the LEDs are precisely located at the edge of the objective numerical aperture ( NA) in the frequency space. Herein, we report a high-speed FPM technique based on programmable annular illuminations (AIFPM). Moreover, the underlying theoretical mechanism as well as optimum illumination scheme for high-accuracy phase imaging in FPM remains unclear. However, the large dataset requirement in conventional FPM significantly limits its imaging speed, resulting in low temporal throughput. Among different approaches, Fourier ptychographic microscopy (FPM) is probably the most promising technique to realize high-throughput QPI by synthesizing a wide-field, high-resolution complex image from multiple angle-variably illuminated, low-resolution images. Blue illumination is advantageous when high temperatures or flames give a strong background radiation of visible or infrared wavelengths or for use with photogenic patterning.High-throughput quantitative phase imaging (QPI) is essential to cellular phenotypes characterization as it allows high-content cell analysis and avoids adverse effects of staining reagents on cellular viability and cell signaling. White illumination is highly efficient and matches any camera‘s sensitivity curve well. The LED 35 Illumination Units contain 9 high power LEDs each and are focused for efficient usage of the light. They come with individual power supplies and are triggered or modulated by TTL signals up to 100 kHz. The LED 35 Illumination Units can be used in a „quasi-CW“ operating mode (light pulses are perceived by the human eye as flicker-free light) or synchronized to other devices and events by DaVis software. LaVision offers LED solutions for different purposes. thermal mechanical fatigue (MFT) testing.background suppression in bright or hot environments.Known spectral characteristics together with low heat dissipation and the ability to work in pulsed mode come in a cost-efficient package for various fields of applications. LED illumination has clear advantages over the alternatives. A wide range of imaging applications do not require the unique illumination capabilities of a laser but still demand for more precise control than a constant halogen illumination can offer.
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