Abstract
Cell imaging using low-light techniques such as bioluminescence, radioluminescence, and low-excitation fluorescence has received increased attention, particularly due to broad commercialization of highly sensitive detectors. However, the dim signals are still regarded as difficult to image using conventional microscopes, where the only low-light microscope in the market is primarily optimized for bioluminescence imaging. Here, we developed a novel modular microscope that is cost-effective and suitable for imaging different low-light luminescence modes. Results show that this microscope system features excellent aberration correction capabilities and enhanced image resolution, where bioluminescence, radioluminescence and epifluorescence images were captured and compared with the commercial bioluminescence microscope.
Highlights
The development of highly sensitive photon sensor arrays has opened new research avenues for probing cell biology via luminescent reporters
While our goal is to evaluate the two tube lenses, it must be noted that the LV200 and the low-light microscope system (LLM) system are equipped with two different Electron multiplying charge-coupled device (EMCCD) cameras (ImageEM C9100-14 and C9100-13, Hamamatsu, respectively)
It should be noted that the magnification is not conventional if the microscope objective – tube lens combination is from different manufacturers
Summary
The development of highly sensitive photon sensor arrays has opened new research avenues for probing cell biology via luminescent reporters. The two most common low emission luminescence signals are bioluminescence (emission of light due to endogenous enzymatic reactions) and chemiluminescence (emission of light by exogenous sensing probes), where luminometer-based assays have been developed to measure bio- or chemi-luminescence for sensitive detection of analytes in bulk samples [4]. Similar to bio- or chemi-luminescence detection, radioluminescence detection is the basis for liquid scintillation counting, which is used to quantify the presence of a radiolabeled probe in a sample [5]. While these bulk assays are very sensitive, they do not have the spatial resolution to distinguish between individual cells or cell compartments
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