Migrasome is a type of recently discovered organelle that plays a vital role in the release of cytosolic contents, regulation of zebrafish embryo formation, mitochondria quality control process, etc. Fluorescence microscopy is widely used to investigate biological specimens, including migrasomes. However, the labelling of fluorescence probes not only requires additional preparation steps, but also may interfere with cellular functions and potentially result in phototoxicity, while only a limited number of labelled structures can be observed at one time. Optical diffraction tomography, as a label-free imaging technique complementary to fluorescence imaging tools, is able to characterize the biophysical properties of organelles. Here we propose to apply optical diffraction tomography for three-dimensional (3D) imaging of migrasome and monitoring its dynamics in living cells.
We have recently demonstrated a high throughput three-dimensional (3D) image flow cytometry method, in which a machine-learning algorithm is used to retrieve the 3D refractive index maps of cells from one angle-multiplexing interferogram. Using this system, we have imaged flowing red blood cells and NIH/3T3 cells with a throughput of more than < 10,000 volumes/second. To further demonstrate its potential on cell phenotyping for clinical testing, we plan to apply this platform to image large populations of various cell types and extracting their morphological and biophysical parameters.
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