Bio-carbon-containing single-cell quantitative techniques based on Raman group.

There are three main forms of storage in organisms: polysaccharides, fats and proteins, which together form the material basis of the bio-carbon storage and bioenergy industry.
at present, the identification, characterization and quantification of these three types of high-energy carbon storage molecules in cells is very cumbersome, and it is often difficult to measure the accuracy of individual cells, which limits the screening and modification efficiency of photosynthesis carbon sequestration cell plants.
the single-cell center of Qingdao Institute of Bioenergy and Process ingenuity of the Chinese Academy of Sciences has invented a single-cell fast-checking technology based on the "Raman group" that measures starch, triglycerides, protein content and oil unsaturatedness simultaneously with the accuracy of a single cell, adding a new means to the performance testing platform of cell factories. The research was published online on November 19,
in Biotechnology for Biofuels.
determine the content of starch, triglycerides, and proteins in cells usually require s3 parallel processes, each consisting of complex steps such as cell culture to accumulate sufficient biomass, extract and isolate target compounds from biomass, and quantify target components by specific methods.
these traditional methods follow the "one process to detect a biomolecular" pattern, which is time-consuming and labor-intensive, and difficult to analyze slow-growing or uncultivated cells.
therefore, it is of great value to develop a rapid, low-cost, high-throughput method for simultaneously determining a variety of carbon storage molecules in a single cell.
"Ramanome" is a collection of single-cell Raman spectra of a cell population in a given state.
researchers based on Raman group techniques, based on rheumine algae and micro-ball algae, established a method for simultaneously quantifying the content of starch, protein, triglycerides and lipid insemior in a single cell (see chart).
the screening speed was increased by at least two orders of magnitude because the Raman group could directly skip microalgae cell culture amplification and complete measurements at the second level without destroying the cells.
on this basis, the researchers put forward new concepts such as cumulative diversity index, cumulative content and cumulative heterogeneity, minimum sampling depth and minimum sampling depth and the safest sampling depth, and established the association between the sampling depth of the Raman group and the accuracy of the phenotype measurement, and theoretically guided the selection and optimization of the measurement parameters of the Raman group.
further proposed that 13 specific Raman peak combination of "cell storage carbon spectrum Raman identification number" can be sensitive, reliable, high-throughput surface to identify starch, protein, triglyceride content and oil unsaturated content and other key phenotypes, and distinguish and reveal the cell carbon storage components and their mutual transformation of static and dynamic characteristics.
In addition, in addition, in the liquid suspended culture of living cells, -80 degrees C frozen-preserved wet algae mud and frozen dry algae powder and other different cell preservation state, the measurement results are highly consistent, so the Raman group technology has the application of universal. the leap forward in the field of
synthetic biology depends to a large extent on the breakthrough of the three common technology platforms, genotype design, genotype synthesis and cell phenotype testing.
with the great improvement of genome sequencing and synthesis in flux and cost, the common link of "cell epitope testing" has become one of the "speed limit steps" of artificial cell construction and biocomponent characterization. the Raman group technology proposed by
researchers can realize the quantization, instrumentation and automation of single-cell function identification, sorting, sequencing and culture of this "cell phenotype test" complete process, combined with a series of single-cell flow Raman sorting techniques such as RADS and RAMS, which are invented in the previous period, without marking, non-destructive and rapid identification of theoretically infinite cell phenotypes.
, theRefore, the Raman group is expected to become a new generation of cell functional testing instrument platform with universality and single-cell phenotytype large data type, serving the energy, environment, health, marine, biosecurity and many other applications of synthetic biology research and industry.
.