1 Introduction to High-Throughput Tissue Homogenization Technology

The high-throughput tissue grinder as an important equipment in modern laboratory sample pretreatment has solved the problems of low efficiency and serious cross-contamination faced by traditional grinding methods. This type of instrument can quickly and parallelly process a large number of samples, and has been widely used in life sciences, biopharmaceuticals, agricultural sciences, environmental detection and other fields.Shanghai Bionoon Biotechnology Co., Ltd. has developed theBionoon series of high-throughput tissue grinders, which adopt an innovative vibration mechanism and a variety of adapter designs, and can complete the grinding treatment of192 samples in30 seconds, effectively improving the efficiency of experiments and the reliability of data.

With the rapid development of (genomics, proteomics, metabolomics) omics technologies, researchers have higher demands for the quality and efficiency of sample pretreatment. The degree of sample homogeneity, nucleic acid / protein extraction efficiency, and avoiding cross-contamination have become key indicators to evaluate the effectiveness of sample pretreatment. Traditional manual grinding or a single grinding method is difficult to meet the requirements of high throughput and high quality simultaneously, while the new generation of high-throughput tissue grinders help to ensure the consistency and reproducibility of sample processing through standardized procedures and programmable control, providing a solid foundation for downstream analysis.

This paper will deeply explore the technical characteristics, working principles and solutions of the high-throughput tissue grinder in different fields, providing professional references for laboratory workers to choose the appropriate sample pretreatment equipment and scheme.

2 Working principle and technical features

The high-throughput tissue grinder is based on the principle of high-frequency vibration crushing, which imparts kinetic energy to the grinding balls in the grinding container, causing the grinding balls to impact, friction, and squeeze the samples at high frequency within the container, thus achieving rapid grinding of tissue-like samples. The Bionoon-48 grinder from Shanghai Bionoon Biotechnology Co., Ltd. employs a special vertical and left-right radial vibration design, and the balls in the tank collide with the samples located on the smooth inner wall of the grinding tank with high energy, thus crushing the samples.

2.1 High-efficiency grinding mechanism

This device generates powerful impact and friction forces through three-dimensional vibration patterns (vertical direction and left-right radial vibration), causing the grinding balls to move at high speed within the sample tube. This multi-directional motion ensures thorough grinding of the samples, even for fibrous and tough samples, achieving uniform and sufficient grinding. The vibration frequency can be adjusted within the range of 0-70Hz（0-2100r/min） according to the needs, adapting to different hardness and toughness of sample types.

The grinding effect can be further improved by using multiple small balls of different diameters. When a large number of small balls (such as glass or agate beads) are used, the huge impact and friction between the balls can effectively achieve the disruption of cells, rapidly releasing the cellular contents. This mechanism is particularly effective for the extraction of nucleic acids and proteins, allowing for the obtaining of longer nucleic acid fragments and higher active proteins.

2.2 Design to prevent cross-contamination

Compared with traditional grinding methods, the high-throughput tissue grinder adopts a fully enclosed sample tube design, where samples remain sealed during the grinding process, and can be used with disposable centrifuge tubes and beads. This design ensures that samples are completely contained within the tube, effectively preventing cross-contamination between samples and contamination from the outside world. Additionally, the device's partitioned design and the use of enclosed grinding jars or disposable centrifuge tubes further reduce the risk of cross-contamination.

For applications scenarios with high cleanliness requirements, such as clinical diagnosis sample processing or micro nucleic acid extraction, the equipment can be optionally equipped with a variety of materials of研磨 vessels and grinding balls, including stainless steel, tungsten carbide, agate, zirconia, and polytetrafluoroethylene, etc., to meet the cleanliness requirements of different experimental conditions.

2.3 Intelligent operation and repeatability

The modern high-throughput tissue grinder is equipped with an intelligent operation interface, usually with a 7 inch touch screen that displays the working status, allowing users to operate intuitively. The built-in program controller can precisely set parameters such as grinding time and the vibration frequency of the rotor, helping to ensure the standardization of the operation process and the reproducibility of the results.

After setting the same protocol for the same tissue sample, a highly consistent grinding effect can be obtained, with little batch-to-batch and within-batch variation, ensuring the reliability of experimental data. The equipment has a short working time (usually 15 seconds - 2 minutes), and the sample temperature will not rise due to long-term grinding, avoiding the risk of degradation of heat-sensitive samples.

Table: Overview of the Technical Parameters of the High-Throughput Tissue Homogenizer

3 Typical Application Scenarios

3.1 Animal and plant tissue treatment scheme

For plant tissues (such as roots, stems, leaves, flowers, fruits, seeds, etc.), the high-throughput tissue grinder can effectively crush the cellulose cell wall, releasing the cellular contents. Different grinding strategies can be adopted for the characteristics of different plant tissues: for samples with a high fiber content (such as stems), it is recommended to use grinding balls made of stainless steel or tungsten carbide, and use a high-frequency vibration mode (1500-1800r/min) for long-term (1-2 minutes) grinding; for samples with a high moisture content (such as fruits, petals), then it is possible to use grinding balls made of zirconia or agate, and use a medium frequency (1000-1200r/min) for short-term (30-60 seconds) grinding, to avoid the loss of samples due to excessive liquefaction.

The processing of animal tissues (such as brain, heart, lung, stomach, liver, thymus, kidney, intestine, lymph node, muscle, bone, etc.) requires parameter optimization according to the toughness and fat content of different tissues. For tissues with strong toughness (such as heart, muscle), a strategy of high-frequency vibration combined with multiple small-diameter grinding balls (3-5mm) can be used; for tissues with high fat content (such as brain, liver), it is recommended to grind under low-temperature conditions to prevent fat oxidation and nucleic acid degradation. The processing of hard samples such as bone and hair requires the selection of grinding balls made of high-density materials (such as tungsten carbide) and the use of intermittent grinding mode to avoid excessive heating.

3.2 Microbial Sample Processing Procedure

The cell wall disruption of microbial samples (including fungi, bacteria, yeast, E. coli, etc.) is a key step in molecular biology and microbiology research. The high-throughput tissue homogenizer can effectively break the tough cell wall of microorganisms through high-intensity mechanical disruption, releasing the cellular contents.

For microbial samples such as yeast and fungal hyphae that are difficult to break, it is recommended to use 0.1-0.5mm diameter small glass beads or zirconia beads, with high-frequency vibration (1800-2100r/min) for multiple short-time cycle grinding (usually 3-4 cycles, each 30 seconds), and the sample can be placed on ice to cool between cycles to prevent overheating and degradation of biological macromolecules. For bacterial precipitates, smaller grinding beads (0.1-0.2mm) and longer grinding time (2-3 minutes) can be used to ensure the complete破碎of the cell wall.

3.3 Food and Drug Inspection Application Scheme

In the field of food and drug safety inspection, the high-throughput tissue grinder can be used for sample pretreatment in component analysis and detection, including various foods and tablets, etc. For samples with complex components, such as health food and traditional Chinese medicine, the equipment can achieve full homogenization, which helps to ensure the representativeness of sampling and improve the accuracy and reliability of the detection results.

Pill crushing is a common step in pharmaceutical quality control, and the high-throughput tissue crusher can process a large number of pills in a short time to obtain a uniform powder, which is convenient for active ingredient content determination and dissolution testing. For pills of different hardness, the vibration frequency and crushing time can be adjusted: for ordinary tablets, medium frequency (1000-1200r/min) crushing 30-60 seconds can be used; for coated tablets and sustained-release tablets with higher hardness, higher frequency (1500-1800r/min) and longer time (1-2 minutes) crushing is required.

The sample types in food analysis are diverse, from meat products to grains, from fresh fruits and vegetables to processed foods, all of which can be quickly pre-treated by high-throughput tissue grinders. For food samples with high water content, a strategy of pre-freezing with liquid nitrogen and then grinding can be used to prevent sample adhesion and component degradation during processing.

3.4 Special Sample Handling Procedure

The high-throughput tissue grinder is also suitable for the processing of a variety of special samples, including volatile samples (such as coal, oil shale, coke, wax products), plastic polymers (such as PE、PS、textiles, resins), and environmental samples (such as soil, sediments, sludge, feces), etc.

For volatile components, the equipment supports low-temperature crushing function, which can pre-cool samples and crushing vessels with liquid nitrogen, and complete crushing under low-temperature conditions to effectively prevent the loss of volatile components. The specific operation process is: immerse the adapter with the sample in liquid nitrogen to cool for 1-2 minutes, take it out and move it to the main machine to be quickly fixed, and the crushing can be started without further freezing, saving liquid nitrogen.

For environmental samples such as soil, sediments, etc., which often contain multiple components with different hardnesses, it is necessary to select appropriate grinding parameters according to the analysis target. If the microbial community in the soil needs to be detected, mild grinding conditions (low frequency, short time) should be used to maintain DNA integrity; if the mineral composition in the soil needs to be analyzed, then more intense grinding conditions (high frequency, long time) are required to ensure that the sampleis fully homogenized.

Table: Recommended grinding parameters for different sample types