Comprehensive solution for algal phenotype research (II)

5. Phenotypic Study of Coral Symbiosis

Miguel Costa LealThe photosynthetic capacity and ecological changes of coral symbionts were studied using the FluorCam closed multispectral fluorescence imaging system. The FluorCam closed multispectral fluorescence imaging system can not only perform chlorophyll fluorescence imaging measurements, but also perform vegetation normalization index NDVI imaging and green fluorescent protein GFP imaging (which can also measure other fluorescent proteins). In this study, NDVI imaging reflected the distribution and concentration changes of chlorophyll, GFP imaging indicated the tissue of echinoderms containing GFP like proteins, and chlorophyll fluorescence imaging demonstrated the photosynthetic capacity of corals. Meanwhile, multispectral fluorescence can also be used for secondary metabolomics and disease phenotype studies, as well as other fluorescent protein imaging, such as YFP, BFP, RFP, etc.

Therefore, a single FluorCam multispectral fluorescence imaging system can complete a comprehensive study of algal phenotypes, including photosynthetic phenotypes, NDVI reflectance spectral indices, fluorescent proteins, secondary metabolism levels and distribution.

Left: NDVI multispectral imaging and GFP green fluorescent protein imaging analysis at different stages of coral contraction expansion; Right; Analysis of RGB imaging, chlorophyll fluorescence imaging, and NDVI multispectral imaging of coral surface and vertical profile samples under low light and strong light adaptation conditions, respectively

Schematic diagram of the structure of the customized FluorCam closed multispectral fluorescence imaging system in this study

reference: Leal M C,et al.2015. Concurrent imaging of chlorophyllfluorescence, Chlorophyll a contentand greenfluorescent proteins-like proteinsof symbiotic cnidarians. Marine Ecology, DOI: 10.1111/maec.12164

6. Research on Biological Erosion Prevention and Control

Marble statues are often covered with green or black gray biofilms. This biofilm is a mixture of blue-green algae, algae, and fungi, and gradually erodes marble. In order to protect outdoor marble statues, European research institutions have conducted a large amount of related research work. To evaluate the activity of biofilms and their effectiveness after cleaning, chlorophyll fluorescence imaging technology is undoubtedly the most convenient and direct technique. Marta Mascalchi from the Florence Institute of Applied Physics in Italy uses laser to remove biofilms from marble. The FluorCam portable fluorescence imaging instrument provides the most powerful data for detecting clearance effects and optimizing laser parameters.

Left: Verify the processing effect through chlorophyll fluorescence imaging; Right: At workFluorCamPortable fluorescence imaging device

reference: Mascalchi M,et al.2018. Laser removal of biofilm from Carrara marble using 532 nm: The first validation study. Measurement 130: 255-263

7. Bioenergy development

The first step in developing algal oil producing bioenergy is to obtain high-quality algal species with high biomass. Non photochemical quenching of NPQ represents the energy dissipated by the photosynthetic system of algae without utilization. So reducing NPQ will inevitably increase its biomass. In this study, FluorCam chlorophyll fluorescence imaging system was used to screen two mutant strains of Chlamydomonas with reduced NPQ, and the optimal cultivation conditions were explored through MC1000 8-channel algae cultivation monitoring system. The MC1000 8-channel algae cultivation monitoring system can be considered as a simplified version of the FMT150 algae cultivation and online monitoring system, which can dynamically regulate light, temperature, and light dark cycles, and monitor algae growth dynamics by measuring OD. It is very suitable for multi replicate rapid cultivation of algae.

Left: NPQ curves, different culture conditions, and growth curves of wild-type Chlamydomonas reinhardtii and two mutant strains; Right: MC1000 8-channel algae cultivation monitoring system

reference: Berteotti S,et al.2016. Increased biomass productivity in green algae by tuning non-photochemical quenching. Scientific Reports 6, 21339

8. Optical imaging analysis system for phenotype omics of marine organisms

2019In the year, Ocean University of China equipped the first domestic optical imaging analysis system for marine biology phenotype omics, which includes the following subsystems:

lFKMMulti spectral fluorescence dynamic microscopy imaging system

lFluorCamMultispectral Fluorescence Imaging System

lFluorCamChlorophyll Fluorescence Imaging System

lSpecim IQhyperspectral imager

lMC1000 8Channel algae cultivation monitoring system

The various subsystems of the optical imaging analysis system for phenotype omics of marine organisms

The FluorCam multispectral fluorescence imaging system is an advanced extension product of FluorCam chlorophyll fluorescence imaging technology. It can be used for dynamic imaging analysis of chlorophyll fluorescence, as well as measurement and analysis of MCF multispectral fluorescence generated by long band UV light excitation on plant leaves. It can also be optionally equipped with imaging measurement of steady-state fluorescence such as green fluorescent protein GFP. FluorCam is widely used in research on algal photosynthetic functional genes, stress response, algal ecology, economic algae breeding, and bioenergy development. Among them, MCF multispectral fluorescence is particularly suitable for the overall analysis of secondary metabolites caused by diseases and the study of algal defense mechanisms.

Left: FluorCam multispectral fluorescence imaging system at work; Right: MCF multispectral fluorescence imaging analysis of seaweed infected diseases

The FKM (Fluorescence Kinetic Microscope) multispectral fluorescence dynamic microscopy imaging system is currently the most powerful and comprehensive instrument for studying plant microfluorescence. It is a customized microscopy imaging system based on FluorCam chlorophyll fluorescence imaging technology. FKM has truly made fluorescence imaging technology a probe for studying the mechanism of photosynthesis, enabling researchers to gain a deeper understanding of the process of photosynthesis and the various changes that occur during it at the cellular and subcellular levels in algae and higher plants. It provides the most powerful tool for directly studying the working mechanism of the photosynthetic system in chloroplasts. FKM, as a dual tool for the microscopic study of algal/plant phenotypes and genotypes, has been widely recognized by the academic community and has achieved a large number of scientific research results. The combination of FKM and FluorCam multispectral fluorescence imaging system can comprehensively reflect the impact of stress factors such as diseases on algae at both macro and micro levels.

Chlorophyll fluorescence microscopy image after infection with seaweed disease: left: maximum fluorescence Fm, middle: maximum photochemical efficiency Fv/Fm; 1 is the initial infected area of the disease, 2 is the area where the disease spreads, and 3 is the area that has not yet been infected; Right: FKM system at work

The Specim IQ Hyperspectral Imager is a complete, portable, handheld hyperspectral camera that integrates features such as hyperspectral data acquisition, analysis and processing, and result visualization. All required components are integrated into a compact and lightweight body, with IP level protection and fully automatic operation. The system comes with a rechargeable battery and a replaceable standard storage card. The measured hyperspectral imaging can further explain changes in algal pigment composition, light adaptation, pigment decomposition in the middle and later stages of stress, and damage to cell structure.

Left: Specim IQ hyperspectral imaging instrument; Right: Hyperspectral imaging analysis of different algae

Ocean University of China hopes to conduct extensive phenotypic analysis and research on different genotypes of algae under stress through this system, in order to reveal the mechanism of algal stress resistance and adaptation, and provide theoretical basis for the development of excellent economic algae species. Preliminary research results have been achieved so far.

Ocean University of China uses FluorCamResearch on Multi Spectral Fluorescence Imaging System for Porphyra yezoensisPyropia yezoensisSecondary metabolic response after infection with red rot disease (Tang L, 2019)

reference: Tang L,et al.2019. Transcriptomic Insights into Innate Immunity Responding to Red Rot Disease in Red AlgaPyropia yezoensis. Int. J. Mol. Sci. 20: five thousand nine hundred and seventy

9. AlgaTechAlgae high-throughput phenotype analysis platform

AlgaTech's customized high-throughput phenotype analysis platform for algae consists of a sample automatic transfer system, a spectral imaging station, and analysis software. It can be equipped with imaging workstations such as multispectral imaging, hyperspectral imaging, chlorophyll fluorescence imaging, and multispectral fluorescence imaging. Each workstation can operate independently and monitor algae photosynthesis, pH, and temperature synchronously. Different types of algae cultivation and online analysis systems can also be selected.