Application

Fluorescence

Automate fluorescence assays in any plate format with Reshape and increase throughput

Analysis

Solid or suspension medium

GFP and RFP

Combine bright-field and fluorescence image information

Reshaping Fluorescence Assays

Fluorescence form the foundation of various laboratory applications, including enzymatic assays, transformant identification, protein detection via ELISA, HT-screening of drug candidate libraries, and analysis of protein-protein interactions.

The Reshape platform offers endpoint or continues fluorescence reading of up to 10 microtiter plates or 15 petri dishes simultaneously. In combination with customized and automated data processing, Reshape Technology achieves deeper insights of your research and improves throughput.

Other applications

Bread mold analysis

Analyze time of mold appearance and track mold growth over time on your food product.

Antagonistic assays

Evaluate inhibitory strain properties and inter-species competition through automated growth detection and quantification using Reshape technology

Seed germination

Automatically analyze complex phenotypes like germination rate, root growth, and time-to-germination with Reshape

Radial growth rate

Highly accurate and time-resolved radial growth assays

Automated CFU counting

The best in class solution for colony counting that actually works

Application Study: Screening for increased recombinant protein titer with 21st BIO

Objective

Identify high producers of recombinant protein using classical mutagenesis in Aspergillus oryzae

Introduction

Classical mutagenesis remains a potent method to generate diverse mutations, especially when working with cell factories. This capability enables identification of numerous and less intuitive combinations of mutations, often unattainable with modern targeted approaches. In this study, UV-mutagenesis was employed to generate a mutant library of Aspergillus oryzae containing an expression cassette for fluorescent protein. Continuous assessment of fluorescence signal and identification of strains with improved protein titer was enabled using the Reshape Imaging System (RIS) in combination with automated image analysis.

Results

Mutant Aspergillus oryzae strains display increased fluorescence. Pools of UV-mutagenized A. oryzae spores were plated on induction medium and development of fluorescence was monitored with Reshape technology. AI-powered image analysis facilitated discrimination between enhanced fluorescence stemming from strain improvement and that from confluent growth of overlapping colonies through time course evaluation (Fig. 1).

Figure 1. Mutagenized fungal spores. UV radiation was employed to mutagenize A. oryzae spores. Subsequently, spores were plated on induction medium in 150 mm petri dishes. Growth and fluorescence were continuously monitored using Reshape Imaging System.

Enhanced fluorescent strains identified with Reshape. Replating of selected candidate strains and tailored image analysis allowed identification of colonies displaying enhanced fluorescence. Fungal colonies were detected in bright field and the fluorescence was quantified over time in the corresponding areas (Fig. 2).

Figure 2. Identification of strains with improved fluorescence. Selected candidate strains were replated. The fluorescence was quantified using Reshape Imaging in combination with tailored data analysis to identify colonies with improved protein titer. Plates were incubated for 2 days.

Increased fluorescence correlates to improved protein yield. To correlate the level of detected fluorescence with protein yield, culture supernatants of the parental strain and selected strains (E1 and E2) were examined by SDS-PAGE. In comparison to the parental A. oryzae strain, the derivatives E1 and E2 significantly improved yield of the recombinant protein (Fig. 3). These results demonstrate the effectiveness of mutagenesis in combination with Reshape technology to increase protein titers in industrial cell factories.

Figure 3. SDS-PAGE of protein titer. The parental strain, ‘enhanced’ strain E1 and E2 were cultivated in deep well plates in triplicates. Culture supernatants from each strain were submitted to SDS-PAGE. The position of the fluorescent protein is indicated by the green arrow.

Conclusion

This study validates UV-mutagenesis combined with Reshape Imaging System and tailored image analysis for identification of enhanced protein yield. The results underscore the effectiveness of this combined approach in boosting protein titers in industrial cell factories, offering promising prospects for further advancements in bioengineering applications.

Food & Ingredients

Supercharge your functional ingredient characterization, stability testing, acidification, and other assays.

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Agriculture

Accelerate agricultural R&D with automated Imaging and AI-powered image analysis

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Personal care

Automate testing and analysis of cosmetics and personal care products to ensure product quality, safety, and efficacy.

Industrials

Enable automated high throughput microbial discovery or do library and enzyme screens with much higher resolution.

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