Imaging technology at various scales of spatial resolution is crucial for understanding the physiological and morphological complexity of living organisms. The existing imaging technologies fail to facilitate images of a sample at different magnification levels at a given instant of time. We report the detailed design and instrumentation of an optical imaging system, namely, simultaneous multiple-level magnification selective plane illumination microscopy (with an acronym being given as sMx-SPIM), which addresses one of the technological challenges of imaging biological specimens at different magnification levels simultaneously. The simultaneous magnified views assist in perceiving biological activities occurring over a short period, especially in developmental biology, where the time scales (fraction of seconds) are critical. The proposed imaging system comprises one illumination arm and two detection arms, each of which uses a different magnification to attain multiple magnification levels simultaneously and overcomes the time consumption for changing the objective lens. The system is automated for image acquisition using a custom-built assembly of motion stages and external hardware. Experimental studies are carried out using biological specimens such as Daniorerio and Alliumcepa to validate the home-built sMx-SPIM imaging system at an obtainable spatial (axial) resolution of ∼ 3 to 5 μm.
The improper disposal of textile dyes like Rhodamine B into nearby water bodies and land areas affects living organisms nearby. Here, we explore the effects of rhodamine absorption through the soil into the plant root and how variation in rhodamine concentration affects the growth and development of the plant. The developmental defects over time are verified by nuclear staining the plant cells. Furthermore, we hypothesis that the chemotropic effect seen in plants guides the root away from rhodamine B high concentration regions to low concentration regions. This will in turn help the plant recover from the injury. These minute structural variations are analyzed with the help of our in-house developed dual-arm multi-level magnification light-sheet microscopy (DMx-LSFM) system. The microscope consists of an automated (translating and rotating) sample holding stage, which helps to observe nearby regions by bringing the region of interest into a plane of focus without disturbing the sample.
The cell wall is the hard layer outside the cell membrane, composed of cellulose, and provides strength to the plant body. Moreover, the tensile strength of the cell wall helps in maintaining the mechanical rigidity and expansion of cells. Any deformation to the cell wall will affect the vitality of the plants. Studies reveal the solubility of cellulose in alcohols, and here we report the study of endurance of cell wall when exposed to methanol. We employed a home-built light sheet microscopy system to visualize the time-dependent deterioration of the cell wall due to the intake of methanol by the plant. The studies were repeated for different concentrations of methanol and show a concentration-dependent degradation curve.
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