From cell disruption to homogenization and pulverization of a great variety of biological samples
Biological samples exist in all shapes and sizes: hard bones, tough and fibrous plants, tough and viscous sputum, soft muscles, tumor or liver tissue. Not to mention the millions of cells such as yeast, bacteria or algae, which have to be disrupted for applications such as DNA or RNA isolation or protein extraction. For research in genomics, transcriptomics or metabolomics, all kinds of biological samples are prepared. Sample preparation is the first step of every analytical process. Retsch offers a range of mills and grinders for easy and reproducible pulverization of solid sample materials some of which are also suitable for cell disruption and homogenization of biological sample materials.
Sometimes the preparation and homogenization of biological samples can be as tough as the material itself. The widely used 2 ml single-use Eppendorf tubes are often not large enough to accommodate the whole sample volume; hence, the sample needs to be divided and reunited after the homogenization process which means an additional time-consuming working step in the lab routine. While it is true that usually larger sized grinding jars, e. g. of stainless steel, are available which accommodate the complete sample volume, these have the drawback of requiring cleaning after use.
A solid sample material should always be sufficiently prepared by size
reduction and homogenization before it is subjected to chemical or physical analysis. Care should be taken that the analysis sample fully represents the original material and that the sample preparation process is carried out reproducibly. Only then are meaningful results guaranteed. Most sample materials can be reduced to the required analytical fineness at room temperature by choosing a mill with a suitable size reduction principle (impact, pressure, friction, shearing, cutting).
How to turn a laboratory sample into a representative part sample with homogeneous analytical fineness -
Food occurs in a great variety of consistencies and is often inhomogeneous. Food testing labs require representative samples to produce meaningful and reproducible analysis results. Therefore, food samples must be homogenized and pulverized to the required analytical fineness, ideally with as little time and effort as possible. Furthermore, reliable analytical results can only be obtained if the entire sample preparation process is carried out reproducibly.
The use of pesticides in agriculture makes it possible to plant extensive mono cultures and often leads to substantial yield increases of food and feed crops. Demand and application have grown steadily over the years, leading to increased contamination of the soil due to the toxic nature of pesticides. Soils save the toxins and their decomposition products so that wildlife is also affected by them. Among the undesired side effects are damages to useful plants and insects like bees. The wind carries pesticides to uncontaminated areas such as fields used for organic farming. Rain also transports the chemicals away from their original area of application to waters and groundwater. Although in most cases the limit values for particular pesticides and their decomposition products are not exceeded, the cumulative effect on humans and animals has not been thoroughly investigated so far. The possible accumulation of pesticides in the food chain could be a source of health hazards; therefore strict quality control of soils is indispensable.
The overall procedure of X-ray fluorescence analysis may be divided into three different stages: sampling, sample preparation and the actual spectrometric analysis itself. Of these three, it is usually the mechanical sample preparation that takes up most of the time and will therefore be discussed in this application report.
We often come across fibrous materials in everyday life. The fibers may be subdivided into natural ones like cellulose, hemp or asbestos and artificial ones like polyester or viscose  (figure 1). The artificial mineral fibers comprise crystalline fibers such as carbon fibers and silicon carbide but also amorphous fibers like glass wool or rock wool. Glass-like fibers are commonly used as insulating wool or as an additive in construction materials to enhance stability, toughness and durability.
Joint replacements, especially of hip and knee joints, rank among the most frequent surgical interventions in industrialized countries. One of the major risks of a joint replacement is prosthetic joint infection (PIJ), a bacterial infection at the interface of implant, tissue, and bone.
In 2010, A.-L. Roux et al. published an article titled „Diagnosis of prosthetic joint infection by beadmill processing of a periprosthetic specimen.“ It describes a new diagnosis method of involved microbes, with an impressive documentation rate of more than 83% and, at the same time, a very low contamination rate of 8.7%. The method involves washing the microbes off the tissue samples with 20 ml sterile water and 5 ml glass beads of 1 mm diameter at 30 Hz in a RETSCH
Mixer Mill within 210 seconds.
Reliable and accurate analysis results can only be guaranteed by reproducible sample preparation. This consists of transforming a laboratory sample into a representative part sample with homogeneous analytical fineness. Retsch offers a comprehensive range of the most modern mills and crushers for coarse, fine and ultra-fine size reduction of almost any material. The product range also comprises a wide choice of grinding tools and accessories which helps to ensure contamination-free preparation of a great variety of sample materials. The selection of the correct grinding tool depends on the sample material and the subsequent method of analysis. Different grinding tools have different characteristics, such as required energy input, hardness or wear-resistance.
In the analysis of solid material, the popular adage that “bigger is better” certainly does not apply. The goal is to produce particles that are sufficiently small to satisfy the requirements of the analysis while ensuring that the final sample accurately represents the original material. The “particles” of interest to the analyst generally range from 10 µm to 2mm. Additionally there are many application, where even finer sizes are needed. One example are active ingredients, where it is necessary to grind in the submicron range. Finally for DNA or RNA extraction mechanical cell lysis is well-established.
Materials differ widely in their composition and physical properties. Hence, there are many different grinding principles that can be applied, and this, together with other variables such as initial feed or “lump” size, fineness needed and amount of sample available, results in a wide range of models available to the researcher.
A variety of methods can be used to analyze solid materials. What they all have in common is the necessity to use a representative, homogeneous analysis sample which needs to have a particular fineness, depending on the analytical method used. The size reduction and homogenization of solids is usually carried out with laboratory crushers and grinders.
A faultless and comparable analysis is closely linked to an accurate sample handling. Only a sample representative of the initial material can provide meaningful analysis results. Rotating dividers and rotary tube dividers are an important means to ensure the representativeness of a sample and thus the reproducibility of the analysis. Correct sample handling consequently minimizes the probability of a production stop due to incorrect analysis results. Thus correct sample handling is the key to effective quality control.
The detection of illegal drugs and pharmaceuticals plays a role in various fields, for example in forensic science, road traffic accidents, in competitive sports or at the workplace. Chemical substances can be detected in blood, saliva, urine and in hair. Hair has the great advantage of storing the substances for a long period, which means that detection is still possible several months after consumption of the drug. In addition to the detection of drugs, hair samples are also used for DNA analysis as well as for the analysis of heavy metals and minerals.
Cell disruption of bacteria, yeast, filamentous fungi or microalgae is a standard procedure in basic biological research, applied biotechnology or medical research to get access to nucleic acids (DNA, RNA) or cell proteins. For the isolation of DNA or RNA usually less than 1 ml of cell material is needed. For the extraction of proteins, however, larger amounts of cell suspension are required. A very efficient method of cell disruption is the co called “bead beating” where cells in suspension are mechanically disrupted by glass beads in single-use reaction vials.
Cashmere wool is the best known precious wool. It is won from the cashmere goat which originates from the high mountain region of the same name. Due to its properties such as softness and warmth, cashmere wool gains more and more popularity in the manufacture of clothing. Genuine cashmere is won solely from the goat’s downy hair and must possess a certain hair structure with an exactly defined length and thickness.
Mechanochemistry is a very effective method to carry out syntheses without solvents and by-products. The technical literature describes a great number of reactions where a conversion of 100% is achieved. A precondition for the establishment of mechanochemistry in the industrial sector is the availability of suitable laboratory mills. A decisive factor is that – similar to conventional preparative chemistry – ambient parameters such as pressure and temperature can be documented and monitored. The Planetary Ball Mills and Mixer Mills from Retsch fulfill these requirements.
Why Size Reduction? A reliable and accurate analysis can only be guaranteed by reproducible sample preparation. The "Art of Milling" describes the process of turning a laboratory sample into a representative part sample with homogeneous analytical fineness.