It is well understood in the field of microscopy that the visualization of the finest details was often limited by the ‘intrusion’ of light from out-of-focus planes. Biomedical research has driven the need for microscopes that can resolve very fine detail in three dimensions within intact, and often living, specimens. The use of fluorescence labelling further exacerbates the problem of out-of-focus light because the signal is generated throughout the volume of the sample of a confocal or wide-field imaging. The following paper describes the most prevalent techniques for 3D imaging and summarizes important optical considerations to achieve high-quality images. These measures allow the production of real-time 3D images with biochemistry techniques to localize the fluorescence protein and generate the labelling of the cell, using the combination of spectroscopy with imaging. However, these approaches are usually limited to low-magnification applications, such as dissection, spectral noise density, or cell biochemistry location. Most compound light microscopes produce flat, 2D images because high-magnification microscope lenses have inherently shallow depth of field, rendering most of the image out of focus and at low resolution. A large open aperture in an optical system can capture high-resolution images but yields a shallow depth of field. This paper proposes a low-cost modular method for retrofitting microscopy imaging systems to achieve 3D focus scanning, and it can correct multispectral density readouts of noise in the sensor. Such a system can lead to miniaturizing portable microscope devices capable of scanning hundreds of cell compounds during its growth, with high contrast and resolution, reducing the challenges of complex post-processing image filtering.
|Publication status||Published - 23 Jun 2021|
|Event||European Light Microscopy Initiative - Online|
Duration: 22 Jun 2021 → 25 Jun 2021
|Conference||European Light Microscopy Initiative|
|Period||22/06/21 → 25/06/21|