Bead Mill Homogenizer Accessories

Ocular tissues, ranging from small, soft perfused tissues to larger, tougher collagenous ones, are commonly selected for analysis during preclinical pharmacokinetic studies for ophthalmic drugs. The process of tissue homogenization and subsequent extraction needs to be optimized for proper downstream drug detection from these various matrices. In this study, bead mill homogenizers were selected for use for its efficiency and effectiveness in preparing complete homogenates. This methodology allows for the extraction of Cyclosporine from eight different ocular tissues – cornea, conjunctiva, iris-ciliary body (ICB), sclera, eyelid, lens, retina-choroid, and lacrimal gland. Due to the different phospholipid profiles between the tissues, however, varying amounts of matrix suppression were encountered during method development; to reduce the effect on analyte response, a phospholipid removal plate was used. The developed LC-MS/MS assay is robust, as well as demonstrates analytical reproducibility and efficiency for the quantification of Cyclosporine in the assayed ocular tissues. Acceptable selectivity, matrix factor, and dilution integrity were demonstrated in detail. Similarly, the methodology was flexible enough to allow for reanalysis of over-range samples as well as in posterior tissues with expected lower Cyclosporine levels. For research use only. Not for use in diagnostic procedures.

Bead mill homogenizers enable rapid and efficient lysis of biological materials, including plants via high-impact forces generated from beads in an enclosed tube to dissociate samples. We explore the efficiency of vertical intra-tube bead milling motions of 7mL and 30mL tube and spiral grinding motions of a 50mL tube using the Omni Bead Ruptor Elite unit. Results indicate protein extraction yields were consistent run-to-run, in different tube sizes, and that full homogenization was achieved in less than a minute for all cases using the same homogenization settings within a given sample type. Thus, having one unit to support varying sample tube sizes allows bead milling to be leveraged for rapid and efficient sample homogenization across various sample input sizes/volumes for protein applications.

Bead mill sample homogenization, exemplified by the Omni Bead Ruptor Elite™ homogenizer, is a rapid and efficient method for sample disruption. However, for longer processing, the kinetic energy generated can increase sample temperature to potentially affect heat-sensitive analytes. We evaluated the passive cooling effect of the 7 mL carriage on larger volume samples, with the carriage and buffers pre-cooled to various temperatures. The results showed that using a carriage equilibrated at -80 °C in combination with chilled buffers effectively kept processing temperatures below 15 °C for up to 1 minute of processing time for larger volume samples though for samples that are in 2mL tubes, active chamber cooling is preferred utilizing a cryo cooling unit.

The comminution step in sample prep is critical for the effectiveness of the QuEChERS method, a type of dSPE, for the multiple pesticide extraction and detection. We evaluate the robustness of bead milling in 50 mL tubes for rapid homogenization of various types of certified organic produce, such as strawberries, soybeans, and apples spiked with 17 component pesticide standards at 100 ppb. Full homogenization by visual inspection was achieved in 30 seconds for all assayed samples of varying density with implemented settings. Pesticides were then extracted and analyzed by GC-MS for % recovery.

Milling is a crucial process in pharmaceutical production and process control, used to enhance solubility and stability of drug compounds through particle size reduction. We explored optimizing dry milling parameters to achieve effective particle size distributions, as dry milling is typically preferred for maintaining product integrity and scalability. On the Omni bead mill homogenizer, optimal milling frequency for analgesics ranged from 25 Hz and 30 Hz, with the most significant particle size reduction occurring within the first minute of milling. Extended milling times beyond five minutes showed negligible additional reduction.

The ability to detect pathogens throughout the food supply chain is an important component of food safety research. Just as the adoption of high-throughput methods such as PCR has expedited molecular detection of microorganisms vs more traditional time-consuming and labor-intensive methods, bead milling allows for an efficient and rapid sample prep method in foods, such as spinach. In this application note, both the bead mill and rotor-stator homogenizers were shown to fully process spinach samples spiked with recombinant E. coli while maintaining their viability. Both methods delivered a 76% cell recovery and a PCR detection limit of 1x10-6 CFUs/mL. Thus, the protocol and methods outlined may help expedite pathogen detection without necessitating a longer pre-enrichment step.

Within agricultural and food studies, genomic analysis plays a large role in elucidating insights into genome modifications, breeding studies, pesticide resistance studies, and identification of plant species. Technologies and methods that can help labs optimize their workflows are valuable, especially if they lend themselves to the throughput, scale, and consistency in nucleic acid extractions for downstream assays, such as PCR or NGS. In this application note, bead mill homogenizers are identified as a robust solution for sample preparation of plant tissues such as spinach leaves, offering an efficient alternative for DNA and RNA extraction compared to time-consuming methods like enzymatic digestion, with resulting homogenates being suitable for downstream analysis. For research use only. Not for use in diagnostic procedures.

DNA analysis of skeletal remains is useful in various scientific fields as bones are known to preserve DNA longer than soft tissues. The extraction of genetic material from bones, however, is challenging due to the degradation of DNA over time and under different environmental conditions. We describe uses of a combination of chemical and mechanical methods to extract DNA from aged Bos taurus bones, focusing on overcoming the low yield of genetic recovery and preventing further degradation compared to fresh tissue.

Plant biology research helps our understanding behind complex mechanisms of plant growth, development, and response to environmental conditions. It touches various aspects from molecular biology, genetics, and ecology through applicable areas in agricultural production, GM crop research, and natural extracts for pharmaceuticals or cosmetics. Within sample preparation for genetic or genotypic analysis, raw sample homogenization is a critical first step to nucleic acid extraction. Bead milling facilitates the disruption and homogenization of various tough plan materials and easily scales for labs looking for high-throughput solutions. In our study, the Omni Bead Ruptor bead mill homogenizer was used to homogenize and extract DNA from frozen spinach leaves and okra pods. Both sample types – soft and fibrous plant tissue – were sufficiently homogenized in a one minute cycle at 4 m/s to take into DNA extraction, resulting in quantities of gDNA cleanly visible by 1D gel electrophoresis, and indicative of high DNA integrity suitable for downstream genomic applications. For research use only. Not for use in diagnostic procedures.

Plants are essential to our diet and ecosystem; researchers are constantly working to better understand their genetic makeup and diversity. Traditional enzymatic lysis methods to extract plant DNA are time-consuming and can be inconsistent due to the toughness of their cell walls. Physical disruption via bead mill homogenization can be an efficient alternative to both rudimentary enzymatic and manual methods. We review the workflow and determine yields from staple crops such as corn, rice, and wheat in this application note. Automation-enabled solutions help scale throughput while providing consistent and efficient sample preparation and nucleic acid extraction from plant material.

Corn is an important stable crop, not only as a food source for humans and livestock, but also biofuel as well other non-food products. This versability makes corn an importance resource and topic of research globally. Within agricultural science and sustainability, labs are hard at work better understanding the genetic mechanisms behind healthy growth and environmental adaptability, whether in response to factors such as climate change, availability of arable land, susceptibility to disease. With the adoption of high-throughput sequencing assessing plant transcriptomics, labs require sample prep methods that reliably allows the extraction of RNA with high integrity for downstream assays, which can be challenging due to thick polysaccharide cell walls and the layers inherent to certain sample types. Semi-automated tabletop bead mills allow for repeatability only afforded by automation-enabled techniques that feed into automated nucleic acid extractions that scales as your throughput increases. For research use only. Not for use in diagnostic procedures.

In the field of pre-clinical and clinical research, sample sourcing and the realistic quantities of tissues or rare samples available for biological analysis can be a major challenge for researchers. Comprehensive studies typically rely on both genomic expression and proteomic profiling readouts to gain for meaningful insights into biological processes, disease progression, or a drug’s therapeutic value, amongst other scientific questions. Well-plate homogenization is an important tool for high-throughput, rapid tissue disruption via bead-milling in well-plates for downstream genomic and proteomic analysis. In this application note, DNA integrity and the protein repertoire assessed using LC-MS/MS techniques for bottom-up proteomic analysis is reviewed after homogenization from rat brain, heart, and small intestine using the same instrument setting for full sample disruption.

Bacterial research used to be limited by the need to plate or grow them in culture to amplify biological analytes for proper downstream analysis and detection. With the continued advances in genomic and proteomic techniques, however, methods such as qPCR and mass spectrophotometry has positively impacted the applicability of bacterial work within various applications in basic research and pre-clinical drug discovery. To help meet the speed at which analysis is now possible, labs can now leverage bead mill technologies in a 96-well plate format to support high-throughput processing. The Omni Bead Ruptor 96 homogenizer, in conjunction with 96-well plates pre-filled with 0.1 mm ceramic bead media, was evaluated for its efficiency in lysing E. coli and recovery of DNA, RNA, and proteins. The results showed high integrity and concentration of these analytes, suggesting a highly efficient system that can stand alone or be integrated with other high-throughput sample prep workflows for various downstream analyses.

Field sampling of mosquito followed by sample dissociation and DNA purification is a routine process for research that surveys the spread of vector-borne diseases. Researchers require a fast and repeatable sample preparation technique to rapidly dissociate a large number of mosquitoes while maintaining a high degree of vector and internal pathogen lysis for quantification of pathogen load. As the specimen sizes are relatively small and the number of samples is high, bead beating is an ideal method for high-throughput sample disaggregation. Herein, we demonstrate dissociation of Aedes aegypti using the Omni Bead Ruptor 96 bead mill homogenizer for the dissociation of mosquitos in both well-plates and tube format and assessment of the extracted DNA from homogenates by endpoint PCR.

Application note on Bead Ruptor 96 homogenization of hemp flower for cannabinoid analysis using HPLC-UV

Bats are essential to the biodiversity critical to sustaining ecosystems, and at the same time, they are also widely studied because they are capable of transmitting viruses to the general human population. Both ecological and animal scientists and human disease researchers depend on reliable sample processing solutions such as bead mills to process tough samples in a safe manner. We evaluate protocols utilizing bead milling in sealed tubes, which helps minimize both risk of dispersion of potentially hazardous materials and risk of cross-contamination without diminishing efficiency, via the Omni Bead Ruptor Elite bead mill homogenizer to process bat fur and skin via both dry-grinding and wet-grinding methods.

Halal and Kosher dietary laws prohibit the use of products containing porcine origin. Pork-based additives, such as collagens used in gelatin or as gelling agents in various processes, are surprisingly common in a variety of food and pharmaceuticals. Bead milling, such as with the Omni Bead Ruptor Elite, offers a high-throughput solution for homogenizing samples for DNA or protein extraction. Methods such as PCR or LC-MS/MS are conductive and crucial for detecting porcine-derived collagen. Bead milling using single-use, sealed tubes are particularly effective for challenging confectionery matrices, and requires no hazardous cleaning solutions; it also offers an alternative that significantly reduces processing time compared to single-sample, manual methods. Herein, we provide recommended parameters for bead mill homogenization of candy.

Sample homogenization is essential in preparing hemp and cannabis samples for cannabinoid analysis. 96-well plate homogenization is a quick and robust method that can be optimized for hemp and cannabis, as it allows for simultaneous processing of multiple samples without the need for cleaning between runs. This method also offers benefits compared to traditional homogenizers, significantly reducing solvent use and increasing throughput, making it a reliable choice for particle size reduction in cannabinoid quantification.

Metagenomics and microbial studies sample prep workflow flyer

Cannabinoid quantification is a common analytical method used by many cannabis producers and testing facilities to define the quantity of cannabinoids in their plants and plant-derived products. The potency of cannabis is typically expressed as the sum of acidic and neutral forms of cannabinoids, with the plant primarily synthesizing the carboxylic acid forms. These forms, however, convert to their neutral counterparts when exposed to substantial heat, such as in the process of smoking or general cannabinoid-containing food production. As such, it is important to maintain this ratio of neutral/acid cannabinoid forms during the sample prep process through analysis. Common methods include manual hand-grinding, rotor-stator homogenization, as well as bead- or cryo-milling. In this study, the effect of bead milling on sample temperature and acidic cannabinoid decarboxylation was evaluated. Cannabinoid standards for THC, THCA, CBD, and CBDA were prepared and serially diluted into a series ranging from 250 ppm (stock) to 7.81 ppm. Samples were analyzed using reverse phase HPLC, where peak areas were integrated for each standard and compared against known concentrations. The degree of decarboxylation was assessed using relative analyte concentrations, and relative amount expressed as a percentage. Results from the parameters evaluated indicate that the Omni Bead Ruptor Elite homogenizer processes cannabinoids without causing decarboxylation, maintaining the ratio of neutral/acid cannabinoid forms for proper quantification

In biomedical research, the ability to reliably extract DNA and RNA from fresh tissue samples is important for molecular applications. It provides an efficient alternative to traditional and manual methods, reducing total processing time and helping with sample-to-sample consistency. We demonstrate the use of the Omni Bead Ruptor Elite™ bead mill homogenizer with stainless-steel lysing/grinding cylinders to yield robust concentrations of DNA and RNA, suitable for downstream molecular applications. The stainless-steel lysing/grinding cylinders are durable and can also facilitate cryogenic sample preparation.

In this study, the Omni Bead Ruptor Elite homogenizer was integrated into a semi-automated workflow for high-throughput nucleic acid extraction from various mouse tissues, including liver, kidney, heart, skin, skeletal muscle, and lungs. An efficient protocol using off-the-shelf chemistry was developed and our results demonstrate its robust scalability, where a processing range from 10 – 100 mg showed proportional increases in nucleic acid yield for most tissue types with this bead mill and KingFisher™ Flex system combination. While liver and kidney samples showed some saturation effects at higher masses, the homogenization parameters and the extraction protocol employed, generally produced nucleic acids that could be reliably feed into common downstream applications such as PCR and sequencing. The workflow, therefore, offers a reliable solution and path to automation for researchers working with multiple types of complex tissue and looking to improve laboratory efficiency. For research use only. Not for use in diagnostic procedures.

The QuEChERS method, a solvent extraction technique introduced in 2003, is widely used for the extraction of multiple pesticides from samples; It has since been optimized to support a broad range of analytes. It starts with the comminution of samples, which can include the convenient and reliable use of bead-beating in solvent. This study describes the application of bead mill homogenization to simultaneously homogenize several samples for QuEChERS extraction for pesticide detection in common fruits and vegetables, with full homogenization achieved in 30 seconds for all assayed produce types in 50 mL tubes. Pesticides were then extracted and analyzed by LC-MS/MS.

Application note quantifying immune mediators from serum and skin tissue using LPS-induced murine model of inflammation via multi-sample processing.