Intracellular heterogeneity is crucial to both cellular pathophysiology and physiology in some crippling illnesses. This is significantly influenced by different organelle populations, and methods that can extract and differentially study organelles from specific places within tissues are needed to understand disease an aetiology. We describe the creation of a subcellular biopsy technique that makes it simpler to separate organelles from human tissue, like mitochondria. We contrasted laser capture micro dissection (LCM), the industry standard for separating cells from the tissues around them, with subcellular biopsy technology. We first demonstrate that LCM has an operating limit of (>20 m) and then demonstrate that subcellular biopsies can be used to isolate mitochondria in human tissue past this limit.

Many human diseases, including cancer, cardiovascular disease, metabolic disease, neurodegeneration, neurodevelopmental disorders, and pathological ageing, are hypothesised to be influenced by inter-tissue and inter-cellular heterogeneity. But analysing heterogeneity at the cellular and tissue levels frequently makes subtle subcellular and organelle heterogeneity difficult to see. As a result of their own multi-copy genome, mitochondria display genetic heterogeneity in addition to the morphological and functional variability seen in other organelles. Homoplasmy, the state in which all mitochondrial DNA (mtDNA) molecules are uniformly wild-type at birth in healthy individuals, is contrasted with heteroplasmy, the state in which de novo mutations produce a mixture of wild-type and mutant mtDNA molecules.

Low levels of heteroplasmy can be tolerated, but if enough mutant mtDNA molecules accumulate and spread, oxidative phosphorylation can become inhibited, which frequently results in mitochondrial illness. This procedure—known as clonal expansion—has an unidentified mechanism. Investigating clonal growth at the subcellular level may aid in characterising mitochondrial illness and aid in our understanding of the mechanisms behind it. In general, a deeper comprehension of the physiological (and pathological) significance of intracellular organelle heterogeneity with subcellular specificity will probably help with efficient illness diagnosis and therapy; however, we need the right technology to do this.

In order to fully benefit from single-cell multiomics, nano probe-based technologies can get beyond some of the problems that are frequently encountered while analysing subcellular molecules, such as: Scanning probe microscopy is frequently used with nano probe technology to achieve nanometer-level accuracy within and outside of cells. Since the relatively small probe size has no effect on the viability of cells or the cellular environment, it permits sampling from living cells. Scanning probe microscopy is frequently used with nano probe technology to achieve nanometer-level accuracy within and outside of cells. Since the relatively small probe size has no effect on the viability of cells or the cellular environment, it permits sampling from living cells.

Here, the coworkers created nano biopsy technique in 2014, which aspirates mitochondria and mRNA from cultivated fibroblasts' cytoplasm using a nanopipette filled with an organic solvent. This approach is based on a process called electro wetting, which involves applying a voltage to a liquid-liquid interface to aspirate a target from the cytoplasm of a living cell. The cytoplasmic proteins and nucleic acids of cultivated cells have recently been successfully sampled using Fluid Force Microscopy (FFM), di electrophoretic nano tweezers, and nano pipettes.

The cytoplasmic proteins and nucleic acids of cultivated cells have recently been successfully sampled using Fluid Force Microscopy (FFM), di electrophoretic nano tweezers, and nano pipettes. The investigation of tissue samples utilised in clinical and molecular pathology hasn't, however, been done using any of these technologies. This study set out to determine whether nano biopsy might be modified to sample from human tissue samples. By comparing subcellular biopsy, a modification of nano biopsy, to Laser-Capture-Micro Dissection (LCM), the most popular technique for examining individual cells in tissue samples, we demonstrate that subcellular biopsy has the potential to move above current methodological limitations