Researchers in many fields need to extract molecules of interest from samples for further analysis. A critical and early step in the isolation procedure is tissue homogenization or cell disruption, the process in which cells are lysed to release intracellular analytes, such as nucleic acids, proteins, or metabolites. These molecules can be analyzed using a variety of techniques, including PCR, NGS, and western blot. As these analytical methods all rely on efficient and reproducible sample preparation, optimizing this step is a priority.
Challenges with existing homogenization methods
The most traditional homogenization technique involves the mechanical pulverization of tissue samples using a mortar and pestle. However, this procedure is time-consuming, laborious, and may not be optimal depending on the physico-chemical properties of the sample.1 Non-mechanical methods, including chemical lysis and sonication, can also homogenize samples. However, the heat generated by sonication and the detergents used in chemical lysis can be harsh on fragile molecules, such as RNA.1,2
More recently developed mechanical methods reduce the labor and time required for lysis, as well as increase the efficiency of homogenization. For example, 3D bead beating technology uses rapidly accelerated bead motion to grind tissue samples.1 The beads inside the lysis tubes are agitated by a multidirectional oscillating movement. Moreover, a large assortment of lysis kits is available and offers the possibility to homogenize any type of biological sample. These kits include tubes whose volume varies from 0.3 to 15 ml, as well as balls made of ceramic, glass or stainless steel, whose diameter varies.
Problematic sample properties
In addition to the challenges associated with the chosen lysis method, the properties of a sample can also hinder analyte extraction. Some samples produce only a small amount of analytes, such as sperm DNA. Consequently, researchers must process larger sample volumes, which take longer to homogenize and may exceed the maximum volume limits of the instrument. Additionally, some samples, such as seeds, skin, bone, or tumor-derived tissues, are difficult to homogenize due to their hardness, elasticity, mineralization, or fibrous nature.1,3–5 To further compound the problem, hard-to-lyse samples generate more heat, leading to the degradation or denaturation of some temperature-sensitive analytes.
Bertin Technologies
The solution to homogenize difficult samples
THE Precellys® Evolution Touch is a versatile homogenizer from Bertin Technologies. It uses bead beating technology, which is the gold standard in tissue homogenization, to grind even the most difficult samples, such as teeth and hair, in minutes. It is able to process samples of different sizes, weights and volumes. In addition, it can homogenize up to twenty-four samples simultaneously. As the symmetry of the instrument allows all tubes to follow the same motion, it ensures equal homogenization efficiency for each sample. The Precellys® Evolution Touch is equipped with a responsive touch screen even when the user is wearing gloves. Additionally, researchers can choose from thirty different lysis kits designed to homogenize samples of varying properties and volumes. With speeds of up to 10,000 rpm, minimal heat is generated through samples, ensuring the safety of temperature-sensitive analytes. In addition, the instrument is compatible with the Cryolys Evolution refrigeration unit, which allows it to maintain the sample temperature between 0 and 10°C before and during homogenization.
Based on the variety of accepted tube sizes, numerous lysis kit options and the ability to process multiple samples simultaneously, the Precellys® Evolution Touch is a versatile and efficient homogenizer suitable for many research areas.
The references
Pagani S, Maglio M, Sicuro L, Fini M, Giavaresi G, Brogini S. Extraction of RNA from cartilage: problems, methods, advice. Int J Mol Sci. 2023;24(3):2120. do I:10.3390/ijms24032120
Shehadul Islam M, Aryasomayajula A, Selvaganapathy PR. A review of macro and micro scale cell lysis methods. Micromachines. 2017;8(3):83. do I:10.3390/mi8030083
Zelentsova EA, Yanshole VV, Tsentalovich YP. A new method for homogenizing samples using a microtome-cryostat device. RSC Adv. 2019;9(65):37809-37817. do I:10.1039/C9RA06808B
Reno C, Marchuk L, Sciore P, Frank C b., Hart D a. Rapid isolation of total RNA from small, hypocellular and dense connective tissue samples. Biotechnics. 1997;22(6):1082-1086. do I:10.2144/97226bm16
Duressa D, Rauscher G, Koike ST, et al. A real-time PCR test for the detection and quantification of Verticillium dahliae in spinach seeds. Phytopathology. 2012;102(4):443-451. do I:10.1094/PHYTO-10-11-0280
