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Microbial 16S/18S FISH Probe Design Synthesis and Detection Service

Microbial 16S/18S FISH Probe Design Synthesis and Detection Service

Overview

CD Genomics specializes in the design and synthesis of custom, single-stranded nucleic acid probes, labeled with fluorescence for specific rRNA sequences. These probes are meticulously engineered to bind only to the target microorganism's rRNA sequence based on complementary pairing, ensuring high specificity. This technique allows for the precise labeling and visualization of microorganisms in various environments, including complex ecological samples, with high spatial resolution under a fluorescence microscope.

Utilizing cutting-edge FISH technology, CD Genomics offers both qualitative and quantitative analyses, allowing researchers to identify individual microbial species or entire populations with unmatched accuracy. This method is particularly beneficial in diverse research areas, including environmental microbiology, clinical diagnostics, and industrial microbiology, where spatial distribution and detailed microbial profiling are critical for advancing scientific understanding.

Advantages of Microbial FISH Detection

High Specificity: allows for precise detection of target microorganismst; enables FISH to accurately identify target strains even in complex samples.
Visualization: enables the direct observation of the expression and distribution of target sequences within cells or tissues.
Safe and Non-contaminating: it is safer to operate and carries no risk of contamination.
Multiplex Detection: simultaneously detect multiple targets using probes with different fluorescence labels.
Broad Applicability: applicable to a wide range of sample types, including bacterial cultures, soil, water samples, plant roots, and intestinal samples.

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Principle of Microbial FISH Probe Design

The core principle of FISH technology relies on the use of highly specific nucleic acid probes that bind to the rRNA sequences of target microorganisms, with fluorescence markers emitting a detectable signal. At CD Genomics, we ensure optimal specificity by utilizing advanced bioinformatics tools and proprietary databases to design probes targeting 16S or 18S rRNA sequences. This precise probe design guarantees unmatched accuracy in detecting and visualizing microorganisms.

Factors Affecting Microbial FISH Detection

To ensure the accuracy and sensitivity of FISH detection, the following factors may influence the experimental results:

Sample Quality

Factors such as cell viability, nucleic acid integrity, and the thickness of tissue sections can all affect the results of the FISH experiment. Ensuring the quality of the sample is crucial.

Probe Quality

The specificity, storage conditions, and concentration of the probe can all affect the experimental outcome. We ensure that all probes are rigorously designed and validated.

Hybridization and Washing Conditions

The temperature, duration, and pH of the washing solutions need to be strictly controlled to reduce non-specific binding and enhance signal specificity.

Microscope Resolution

The resolution and sensitivity of the fluorescence microscope will directly impact the detection of fluorescence signals. We use high-quality microscopy equipment to ensure result accuracy.

How to Choose the Fluorescent Label for FISH Probes?

Fluorescent Label Application	Description
Single Bacterial 16S Label	Recommended to use Texas Red, compatible with CY3. FITC can be used as a negative control.
Dual-Color FISH Label	Used for detecting two different strains or using two different fluorescent labels for the same strain. FITC is recommended as the positive control, and the target microorganism can be labeled with Texas Red or CY5.

Combined FISH Detection and Antibody Detection

FISH technology can be used alone or in combination with antibody detection for joint experiments. This allows you to simultaneously detect nucleic acids and proteins in the same experiment. This method is particularly useful for studies that require observing both the gene and protein expression of microorganisms. It is important to carefully optimize factors such as the choice of fixative, avoidance of nucleases, and compatibility of fluorescent labels to ensure that the detection results of the two methods do not interfere with each other.

Service Specifications

CD Genomics Customized FISH Detection Services

1. Probe Design and Synthesis: Based on your research goals, we provide precise probe design and synthesis services.

2. Experimental Condition Optimization: We optimize the experimental conditions according to the sample type and characteristics of the target microorganism to ensure the best experimental outcome.

3. Technical Support: We offer detailed experimental operation guidance and technical support to ensure your experiments proceed smoothly.

Data Analysis and Result Interpretation: After the experiment is completed, we provide a comprehensive results analysis report and assist you in interpreting the experimental data.

Why Choose CD Genomics' 16S FISH Probe Design Service?

With its strong technical background, extensive database resources, and years of experimental experience, CD Genomics is capable of providing high-quality, customized FISH probe design services to clients. Here are several unique advantages of our services:

High-Quality Probe Design: Leveraging the world's largest 16S database, we evaluate millions of 16S sequences and select the most specific probes to ensure high-precision detection.
Personalized Customization: Based on your research needs, we can design probes tailored to specific targets, whether for a single microorganism or a complex microbial community. CD Genomics provides the best solutions for your specific research objectives.
Comprehensive Technical Support: From experimental planning and probe selection to data analysis and result interpretation, we provide full technical support to ensure that your experiments are successful and that the data is accurate and reliable.
Multiplex Probe Design: For research that requires simultaneous detection of multiple microbial targets, CD Genomics offers multiplex FISH probe design services, supporting the simultaneous hybridization of probes with different fluorescent labels.

Microbial FISH Detection Workflow

Probe Design and Synthesis

We begin by designing and synthesizing highly specific probes tailored to the 16S or 18S rRNA sequences of your target microorganism. The probes are rigorously tested for specificity, ensuring they bind exclusively to the desired microorganism's rRNA.

Sample Fixation

Proper fixation preserves the integrity of the microorganism's cellular structure and nucleic acid, ensuring that the probes can effectively enter the cells. Samples may include bacterial cultures, tissue sections, and environmental samples like soil or water.

Sample Permeabilization

Samples undergo a permeabilization process, which is optimized to enhance probe entry and binding specificity, ensuring the probes reach their intended target within the cells.

Overnight Hybridization

Probes are incubated with the sample overnight under controlled conditions to allow sufficient time for hybridization with the target rRNA sequences.

Washing

Excess probes and non-specifically bound probes are removed through a washing step, reducing background signals and enhancing the clarity of the results.

Fluorescence Microscopy Observation

The sample is then analyzed under a fluorescence microscope, where the detected fluorescence signals indicate the presence and distribution of the target microorganisms. By using different fluorescence markers, we can capture multiple data points, including the abundance and spatial distribution of the target microorganism.

Interpretation of Microbial FISH Detection Results

Result Type	Description
Positive Result	Observed the expected fluorescence signal, indicating the presence of the target sequence and successful detection of the target microorganism.
Negative Result	No fluorescence signal observed, indicating that the target sequence is absent or its expression level is below the detection limit.
Non-specific Binding	Background fluorescence observed, suggesting that the probe may have bound to non-target sequences, requiring optimization of experimental conditions.

Samples Requirement

To ensure that we provide the most accurate FISH probe design and detection services, we require the following information from you:

Sample Type: This can include cell cultures, tissue sections, microbial community samples, etc.
Target Microorganism's Genetic Sequence or GeneID: This is the basis for designing specific probes and helps us accurately locate the target sequence.

CD Genomics' Species-Specific 16S FISH Probe Design Service provides precise and reliable molecular tools for your microbial research. By utilizing advanced probe design platforms, extensive database support, and rigorous experimental protocols, we ensure that each client receives highly specific and sensitive experimental results. Whether you are conducting microbial community analysis, pathogen detection, or ecological research, CD Genomics can offer the most suitable solutions for your needs. If you have any questions or require further details about our services, please feel free to contact us. We are dedicated to providing professional technical support and service.